Development of a Squid-hydrolysate-based Larval Diet and its Feeding Performance on Summer Flounder, Paralichthys dentatus, Larvae

Locally generated squid‐processing byproduct was processed into concentrated hydrolysate (22% solids, 17.3% protein, and 3.0% lipid, primarily phospholipids—11.6% eicosapentaenoic acid/24.5% docosahexaenoic acid on a lipid weight basis). Two microparticulate diets (65% protein, 19% lipid, 7.5% carbohydrate, and 19.12 MJ/kg energy, on a dry weight basis) were prepared using squid hydrolysate (SH) and squid‐herring hydrolysate as sole protein sources (73.3 and 78.65% of the whole diet, respectively). A 22‐d feeding trial with summer flounder, Paralichthys dentatus, larvae of 17 d after hatch showed that the survival rate (92%) of larvae fed SH was significantly (P < 0.05) higher than those of larvae fed live Artemia nauplii (81%) and a commercial diet, Proton (65%), while specific growth rates (SGR) were comparable (2.23% /d for SH and 2.86% /d for Artemia) with the lowest for Proton (1.39% /d). After switching from commercial and Artemia diets to a SH diet for 17 d following the 22‐d feeding, significant improvements were seen in survival rates of postweaning larvae fed previously commercial (65.28–76.57%) and Artemia diets (81.25–89.07%).     

Channel Catfish,Ictalurus punctatus,Immunity to Saprolegnia sp.

ABSTRACT

The Japanese flounder, Paralichthys olivaceus, is one of the most common finfish cultured in Japan and Korea. Despite the relatively high production of fingerlings, some problems remain, mainly related to the larval feeding and cost of maintaining microalgae and rotifers. In order to determine the effects of different diets on the Japanese flounder larval growth and survival, a series of experiments was carried out related to the size and nutritional value of different live feeds. The larvae culture conditions were at 10 or 20 larvae/L in 50 to 2,000 L tanks, with aeration and with or without “green water,” and a temperature range of 18.5 to 22.5°C. The live foods used were microalgae (Chlorella ellipsoidea and Nannochloris oculata), baker's yeast, experimental n-yeasts, oyster trochophore larvae, three strains of rotifer Brachionus plicatilis (L-type, S-type and U-type) and Artemia nauplii. Variations were detected in size, dry weight, and chemical composition of the three strains of rotifers used. The maximum number of rotifers ingested by flounder larvae increased steadily from 7 individuals, at first feeding (3.13 mm), to 42 individuals at 5.25 mm of total length (6 days after first feeding). There was a relationship between larval total length and size of the rotifers ingested. The effect of rotifer size on larval growth and survival appeared to be limited to the first two days of feeding. Of the diets tested in the growth and survival of larval flounder during 14 days after hatching, rotifers fed on C. ellipsoidea and raised in green-water gave the best results. Rotifers cultured on enriched N. oculata and n-yeasts did not support larval growth and caused higher mortalities. The n-yeasts used as rotifer enrichment appeared to satisfy, partially, the nutritional requirement of 7-day-old flounder larvae, as did n-yeast squid wintering oil the requirements of 14-day-old larvae. From 7-9-days after hatching and throughout the second 14-day period, rotifers and Artemia cultured on N. oculata improved the survival of flounder compared with those fed on rotifers cultured on C. ellipsoidea. Moreover, the larval growth did not vary significantly between both microalgae-rotifer feedings. No clear relation was found between total protein, lipid, amino acids and fatty acids of live feeds with the growth and survival of flounder larvae, although the total lipid was higher in C. ellipsoidea than in N. oculata. The Artemia nauplii San Francisco strain appeared to be more suitable for the growth and survival of flounder larvae, than the Utah strain. The nutritional value of Artemia nauplii (Utah strain) for flounder larvae remained unchanged despite the use of either microalgae as nauplii enrichment.     

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.     

Establishing larval feeding regimens for the Forktail Blenny Meiacanthus atrodorsalis (Günther, 1877): effects of Artemia strain,time of prey switch and co‐feeding period

This study aimed to establish feeding strategies covering the whole larval period of the forktail blenny, Meiacanthus atrodorsalis, based on the standard hatchery feeds of rotifers and Artemia. Three purposely designed experiments were conducted to determine the appropriate times and techniques to transition larvae from rotifers onto Artemia nauplii of a Great Salt Lake (GSL) strain, and a specialty AF strain, as well as subsequent transition onto enriched metanauplii of GSL Artemia. With a 3‐day co‐feeding period, larvae adapted well to a transition from rotifers to newly hatched GSL Artemia nauplii as early as 5 days posthatching (DPH), and as early as 3 DPH when fed the smaller AF Artemia nauplii. However, prolonging the rotifer‐feeding period up to 11 DPH did not negatively affect survival. Larvae fed Artemia nauplii of the AF strain showed 17–21% higher survival, 24–33% greater standard length and body depth, and 91–200% greater dry weight, after 20 days relative to those fed nauplii of the GSL strain. Meanwhile, enriched Artemia metanauplii of the GSL strain were shown to be an acceptable alternative to AF Artemia nauplii for later larvae, producing similar survival and growth when introduced from 8 DPH. Based on our findings, we recommend feeding M. atrodorsalis larvae rotifers as a first food between 0 and 2 DPH, introducing AF Artemia nauplii from 3 DPH, followed by enriched GSL Artemia metanauplii from 8 DPH onward, with a 3‐day co‐feeding period between each prey change.     

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.     

Survival and Growth Improvement of Palm Ruff,Seriolella violacea,Larvae Fed Artemia Nauplii Enriched with an Experimental Emulsion

The palm ruff, Seriolella violacea (Cojinoba), is a potential new species for Chilean aquaculture. To approach Cojinoba larviculture, an experimental Artemia enrichment emulsion, containing docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA) = 2.5, supplemented with vitamin E, astaxanthin, and β‐glucan, was evaluated in both Artemia and Cojinoba larvae, 30–50 d.a.h. This study tested an experimental enrichment emulsion versus a commercial emulsion, with an integral approach of multicompound emulsions. After 23 h enrichment, experimental emulsion (EE)‐enriched nauplii reached DHA and EPA concentrations of 23.8 and 18.7 mg/g dry weight (dwt), respectively, while in Cojinoba larvae they were 18.4 and 19.7 mg/g dwt. Control emulsion (CE)‐enriched nauplii exhibited lower DHA and EPA (6.1 and 7.7 mg/g dwt), while only DHA decreased in the control larvae (12.6 mg/g dwt). Vitamin E was higher in EE‐enriched nauplii (29.2 mg/100 g dwt) than in the control (8.4 mg/100 g dwt). Larvae fed EE‐enriched Artemia exhibited 8% increase in survival and 19% in growth compared with the control. Astaxanthin was detected only in larvae fed EE‐enriched nauplii. The tumor necrosis factor‐α concentration was not significantly different between larvae fed EE‐ and CE‐enriched nauplii. EE looks promising as an Artemia enrichment and experimental diet to assess palm ruff larval requirements, and has a positive impact on fish larvae performance.     

营养强化的轮虫、卤虫对牙鲆仔鱼的成活、生长及体脂肪酸组成的影响

用3种营养强化剂强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,研究牙鲆仔鱼的生长、成活、体脂肪酸的组成。结果表明:用强化的轮虫和卤虫无节幼体投喂牙鲆仔鱼,成活率、增重均显著高于对照组(p<0 01),其中V号强化剂的效果最好,成活率为29 34%,比对照组提高100%;增重倍数为217 90,比对照组提高68 61%;这是由于V号强化剂强化的卤虫无节幼体体内含有较多的AA的缘故,饵料中AA含量的提高,可以提高牙鲆仔鱼的成活率、促进其生长。牙鲆摄食强化过的轮虫、卤虫无节幼体后,其EPA、DHA、n-3HUFA、PUFA的含量随着饵料中含量的升高而升高,这也是牙鲆仔鱼生长速度和成活率提高的重要因素之一。     

Effect of enriched live feeds on survival and growth rates in larval Korean rockfish, Sebastes schlegeli Hilgendorf

High mortality frequently occurs in larval mass production of Korean rockfish, Sebastes schlegeli Hilgendorf. Nutritional deficiencies in live feeds, rotifers and Artemia nauplii, fed to larvae could be a reason. A series of experiments was carried out to evaluate the effect of nutritional enrichment of live feeds by ω‐yeast, Spirulina powder and Super Selco^TM on survival and growth rates in rockfish larvae. Preference of rockfish larvae for the live feeds was determined by analysis of stomach contents. In addition, the effect of green water produced by the use of Chlorella ellipsoidea and Spirulina powder on the growth performance of larvae was evaluated. Larvae fed rotifers nutritionally enriched with Super Selco showed significantly higher survival rates than those fed rotifers enriched with ω‐yeast. Larvae fed rotifers that were nutritionally enriched with both Super Selco and Spirulina together exhibited improved growth and survival rates. Larvae fed Artemia nauplii nutritionally enriched with Spirulina powder showed significantly higher survival than larvae fed Artemia nauplii without enrichment. When larvae were fed rotifers, Artemia nauplii or the mixture of rotifers and Artemia nauplii, the second and last group showed significantly higher survival than the first group. Fatty acid composition in live feeds was improved by enrichment of ω‐yeast and larvae fed this feed showed higher survival and growth rates compared with larvae fed non‐enriched feeds. No positive effect of green water in the tank produced with C. ellipsoidea or Spirulina powder was observed on survival and growth rates for larvae fed nutritionally enriched rotifers with Super Selco and Spirulina powder. However, when the larvae were fed Artemia nauplii that were nutritionally enriched with ω‐yeast and Spirulina powder, green water obtained by adding Spirulina powder to the tanks resulted in significantly higher growth rates of larvae than was obtained by adding C. ellipsoidea.     

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 Effects of Feeding on Muscle Growth Dynamics and the Proliferation of Myogenic Progenitor Cells during Pike Perch Development (Sander lucioperca)

The effects of feeding on the development and growth of pike perch muscle and on proliferation of their progenitor myogenic cells were evaluated. Larvae were fed starting on Day 5 after hatching with Artemia nauplii, two commercial diets (Aglo Norse [AN] and Biokyowa [BK]), and two formulated diets (C [nonhydrolyzed casein] and CH [25% casein hydrolysate]). The survival, body mass, and length of pike perch juveniles fed Artemia nauplii and AN and BK diets were significantly higher compared to the C and CH groups. The highest somatic growth rate was associated with an increased contribution of hyperplasia to white muscle growth. Significantly higher frequency of proliferating cell nuclear antigen‐ and Ki‐67‐positive nuclei in the white muscle of fish fed Artemia nauplii and commercial diets compared to those fed C and CH feeds indicates that feeding affected the number of fibers. The pike perch fed the CH diet exhibited significantly lower total cross‐section area and average fiber area, additionally to the pathological changes in muscle morphology. The larvae fed natural food and diets promoting a fast growth rate exhibited a higher contribution of hyperplasia to muscle growth, which in turn, promoted an increase in the body size of adult fish.     

Effects of enriched Artemia nauplii on production, survival and growth of the mysid shrimp Mysidopsis almyra Bowman 1964 (Crustacea: Mysidacea)

The effects of enriched Artemia nauplii on larvae production and survival and growth of the mysid Mysidopsis almyra Bowman 1964 are compared. There were no significant differences (P > 0.05) in production between mysids fed the Artemia nauplii (133 ± 69 mysids day⁻¹) and mysids fed the enriched nauplii (139 ± 82 mysids day⁻¹). No differences in size of newly hatched mysids or mysid growth to 15 days (P > 0.05) were found between the two diets. Survival was significantly higher (P < 0.05) for mysids fed the enriched nauplii (59.1%) compared with mysids fed Artemia nauplii (41.4%).     

Effect of Rearing Density and Feeding Regimes on Growth and Survival of African Catfish,Heterobranchus longifilis (Valenceinnes, 1840) Larvae in a Closed Recirculating Aquaculture System

Heterobranchus longifilis larvae were reared over a 35 d period to evaluate the effects of stocking densities and feeding regimes on growth and survival. In experiment 1, larvae (12.3?±?2.1 mg) were stocked into glass aquaria at densities of 1, 2, 5, 10, 15, 20, and 25 larvae L^?1. Larvae were fed on Artemia nauplii ad libitum. Significant variations in terms of growth performance and feed utilization occurred at all levels of density treatments. Specific growth rate (SGR), body weight gain (BWG), and feed efficiency (FE) of the larvae decreased significantly as density increased. However, survival rate increased with the increase of stocking density. In experiment 2, larvae (13.4?±?1.1 mg) stocked at a density of 15 larvae L^?1, in the same conditions as experiment 1, were fed on three different regimes: Artemia nauplii; 35%?protein beef brain; and 35%?protein commercial catfish feed (CN⁺). SGR, BWG, and coefficient of variation (CV) of larvae fed on Artemia nauplii were significantly higher than those fed on beef brain and CN⁺. The survival rate of larvae fed on beef brain was significantly higher (88.40?±?9.75%) than those of Artemia (69.21?±?6.69%) and CN⁺ (40.40?±?6.22%). The results of this study suggest that the optimum stocking density is 15 larvae L^?1 and the beef brain can be used as alternative feed to Artemia in rearing H. longifilis larvae.     

Preliminary study on the activity of protease enzymes in Persian sturgeon (Acipenser persicus Borodin, 1897) larvae in response to different diets: effects on growth and survival

The specific activity of alkaline protease, trypsin and pepsin‐like enzymes was measured in yolk sac stage Acipenser persicus larvae and over a 1‐month feeding experiment using live Artemia nauplii (ND), formulated feed (FD) or mixed food (MD). Artemia nauplii group larvae showed significantly higher growth and survival during the first 15 days while FD larvae showed the lowest growth and survival. At day 30, MD larvae exhibited significantly higher growth than the ND group. Alkaline protease activity showed the lowest activity on day 15; the highest activity was observed in the MD group larvae. Pepsin‐like activity showed a drastic increase from day 1 to 5 in all treatments, but remained stable throughout the next 25 days, with the lowest and the highest activity in the FD treatment on day 10 and in the MD treatment on day 30 post‐feeding respectively. Trypsin‐like activity in group ND remained almost the same from day 5 to 30, whereas in groups MD and FD, it decreased significantly from day 10 to 30. The contribution of the naupliar proteases was moderate but effective. Additionally, better performance in Artemia fed sturgeon larvae may also be due to the structure and digestibility of proteins and the food intake stimulation by the nauplii.     

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

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

Growth, survival, quality and digestive enzyme activities of larval shrimp fed microencapsulated, mixed and live diets

An artificial diet for shrimp larvae was microencapsulated with a polysaccharide blend [66% gum arabic, 17% mesquite gum and 17% maltodextrin 10 dextrose equivalent (DE)]. Microencapsulated diet (MD) was fed to mysis alone, as a co‐feed with the microalgae Chaetoceros cerastosporum and Tetraselmis chuii (mixed) and compared with a live diet (control) of C. cerastosporum, T. chuii and Artemia nauplii. No significant differences (P > 0.05) were detected in growth rates, development and quality indexes of larvae fed the three experimental diets. All diets supported survival percentages of >90%. Shrimp larvae fed MD and mixed diets had higher specific trypsin activity and soluble protein content than those fed live diet. Amylase activity decreased in larvae fed the mixed and MD apparently due to the carbohydrate composition of the diet. The results indicate that it is possible to substitute a live diet with a microencapsulated one (with a wall composition made up of a polysaccharide blend) in Litopenaeus vannamei mysis.     

Effect of Bacillus subtilis on growth development and survival of larvae Macrobrachium rosenbergii (de Man)

A feeding experiment was conducted to investigate the effect of Bacillus subtilis bacterium, on larval growth and development rate of Macrobrachium rosenbergii (de Man) during February 28 to April 8, 2005 in University Putra Malaysia hatchery. Newly hatched larvae of M. rosenbergii were reared with two dietary treatments consisting of newly hatched Artemia salina nauplii with B. subtilis (10⁸ cells ml⁻¹), and newly hatched A. salina nauplii without B. subtilis carried out in triplicate in 60‐L aquarium (50 L⁻¹). After trial, the larvae that fed B. subtilis‐treated Artemia naupli were found to have higher survival and a faster rate of metamorphosis than larvae that were fed with nontreated Artemia naupli. There were significant differences between B. subtilis‐treated Artemia naupli and nontreated Artemia diet in larval growth and development rate of metamorphosis (P < 0.05). Larval survival after 40 days was significantly greater (P < 0.05) in the B. subtilis‐treated groups (55.3 ± 1.02) compared with the nontreated groups (36.2 ± 5.02%).     

Preliminary results of experiments on the rearing of Tasmanian flounders, Rhombosolea tapirina and Ammotretis rostratus

Tasmanian species of flounder were cultured successfully following stripping and fertilizing eggs after hormone-induced ovulation. The larvae were fed rotifers followed by Artemia nauplii. Survival rates of larvae from first-feeding to metamorphosis, which were as high as 94–98% for R. tapirina and 65% for A. rostratus, indicate that both species can be readily cultured in captivity.     

Larval rearing of African carp,Labeo parvus Boulenger, 1902 (Pisces: Cyprinidae), using live food and artificial diet under controlled conditions

This study aimed at (1) evaluating the efficacy of live food organisms (Artemia and natural zooplankton) and an artificial diet in the larval rearing of African carp Labeo parvus and (2) determining appropriate rearing conditions. After yolk sac resorption, the larvae were separated into five different feeding trials with two replicates. At the end of the larval rearing period (28 days post‐hatching), the highest (94.6%) and the lowest (53.7%) cumulative survival rates were found when larvae were fed with natural zooplankton for 7 days followed by Nippai food for 21 days, and when larvae were fed from the beginning of exogenous feeding with Nippai food only respectively. The significant highest body weight (351.6 mg), total length (34.4 mm) and specific growth rates (15.5%day^?1) were recorded when the larvae were fed with Artemia nauplii for 14 days followed by Nippai food for 14 days. The lowest growth performance (body weight and specific growth rates) were obtained when larvae were fed exclusively Nippai food. These results indicate that L. parvus can be successfully cultured in indoor nursery systems from hatching to the early juvenile stage.     

Effects of dietary eicosapentaenoic acid on growth, survival, pigmentation and fatty acid composition in Senegal sole (Solea senegalensis) larvae during the Artemia feeding period

We examined the effect of dietary eicosapentaenoic acid (EPA, 20:5n‐3) on growth, survival, pigmentation and fatty acid composition of Senegal sole larvae. From 3 to 40 days post‐hatch (dph), larvae were fed live food that had been enriched using one of four experimental emulsions containing graduated concentrations of EPA and constant docosahexaenoic acid (DHA, 22:6n‐3) and arachidonic acid (ARA, 20:4n‐6). Final proportions of EPA in the enriched Artemia nauplii were described as ‘nil’ (EPA‐N, 0.5% total fatty acids, TFA), ‘low’ (EPA‐L, 10.7% TFA), ‘medium’ (EPA‐M, 20.3% TFA) or ‘high’ (EPA‐H, 29.5% TFA). Significant differences among dietary treatments in larval length were observed at 25, 30 and 40 dph, and in dry weight at 30 and 40 dph, although no significant correlation could be found between dietary EPA content and growth. Eye migration at 17 and 25 dph was affected by dietary levels of EPA. Significantly lower survival was observed in fish fed EPA‐H diet. Lower percentage of fish fed EPA‐N (82.7%) and EPA‐L (82.9%) diets were normally pigmented compared with the fish fed EPA‐M (98.1%) and EPA‐H (99.4%) enriched nauplii. Tissue fatty acid concentrations reflected the corresponding dietary composition. ARA and DHA levels in all the tissues examined were inversely related to dietary EPA. This work concluded that Senegal sole larvae have a very low EPA requirement during the live feeding period.     

Stable carbon (δ13C) and nitrogen (δ15N) isotopes as natural indicators of live and dry food in Piaractus mesopotamicus (Holmberg, 1887) larval tissue

This study proposed the use of the stable isotope technique to track the type of food utilized by pacu Piaractus mesopotamicus larvae during their development, and to identify the moment when the larvae start using nutrients from the dry diet by retaining its carbon and nitrogen atoms in their body tissues. Five‐day‐old pacu larvae at the onset of exogenous feeding were fed Artemia nauplii or formulated diet exclusively; nauplii+formulated diet during the entire period; or were weaned from nauplii to a dry diet after 3, 6 or 12 days after the first feeding. δ¹³C and δ¹⁵N values for Artemia nauplii were ?15.1‰ and 4.7‰, respectively, and ?25.0‰ and 7.4‰ for the dry diet. The initial isotopic composition of the larval tissue was ?20.2‰ and 9.5‰ for δ¹³C and δ¹⁵N respectively. Later, at the end of a 42‐day feeding period, larvae fed Artemia nauplii alone reached values of ?12.7‰ and 7.0‰ for δ¹³C and δ¹⁵N respectively. Larvae that received the formulated diet alone showed values of ?22.7‰ for δ¹³C and 9.6‰ for δ¹⁵N. The stable isotope technique was precise, and the time at which the larvae utilized Artemia nauplii, and later dry diet as a food source could be clearly defined.