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.     

Senegalese sole larvae growth and protein utilization is depressed when co‐fed high levels of inert diet and Artemia since first feeding

A large effort has been dedicated in the past years to the development of nutritional balanced inert diets for marine fish larvae in order to suppress the nutritional deficiencies of live feed. In this study growth performance, Artemia intake, protein digestibility and protein retention were measured for Senegalese sole (Solea senegalensis Kaup), in order to provide insight into how protein utilization affects growth performance. Three feeding regimes were tested: ST – standard live feed; ArtRL – live feed and 20%Artemia replacement with inert diet (dry matter basis) from mouth opening; ArtRH – live feed and 58%Artemia replacement with inert diet from mouth opening. Artemia intake and protein metabolism were determined at 6, 15 and 21 days after hatching using ¹⁴C‐labelled Artemia protein and subsequent incubation in metabolic chambers. At the end of the experiment, sole fed exclusively with live feed were significantly larger than sole from Artemia replacement treatments. Protein digestibility decreased during sole ontogeny, and more sharply in ArtRH sole. Concomitantly retention efficiency increased during ontogeny but with a slight delay in ArtRH sole. Senegalese sole larvae growth and protein utilization is depressed when co‐fed high levels of inert diet and Artemia, mostly during metamorphosis climax.     

Weaning of the wedge sole <Emphasis Type="Italic">Dicologoglossa cuneata</Emphasis> (Moreau): influence of initial size on survival and growth

The weaning phase can be decisive in fish-culture viability. In this work, the relationship between the initial size and weaning success has been studied in wedge sole (Dicologoglossa cuneata). For each age (30, 50, and 70 days after hatching, DAH), two to three sizes were selected, and all were put on the same feeding schedule for 20 days. Each batch (three replicates) was sampled at 1, 10, and 20 days. Specific growth rate (SGR) and survival were compared at the end of the co-feeding period, after 10 days on dry feed only. The best results for survival and growth were found with the smallest larvae, and vice versa. The SGRs and survival rates recorded during the co-feeding period were higher (0.8–15.6 day⁻¹ and 68.3–97.8%) than those from the dry-food phase (0.9–4.7 day⁻¹ and 56.3–66.7%). Successful weaning (survival = 65% and SGR = 9.3 day⁻¹) is possible with 30 DAH larvae (7.6–8.1 mm and 3.9–4.6 mg). In conclusion, the most effective weaning would be possible at 30 DAH, implying significant Artemia savings (25–50%).     

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.     

Optimum time for weaning South African <Emphasis Type="Italic">Scylla serrata</Emphasis> (Forskål) larvae from rotifers to <Emphasis Type="Italic">Artemia</Emphasis>

To determine the optimum time at which to wean Scylla serrata larvae from rotifers onto Artemia two experiments were conducted, approximately 1 month apart, using larvae from two different female crabs. In the first experiment, the larvae in three treatment groups, with nine replicates each, were fed rotifers for the first 8 days after hatching. Artemia were introduced on days after hatch (DAH) 0 – during the first zoeal instar (treatment R + A); on DAH 4 – during the second zoeal instar (treatment R4A); on DAH 8 – during the third zoeal instar (treatment R8A). In a control (ROT) larvae were fed with rotifers exclusively for 18 days until the completion of metamorphosis to megalopa. In the second experiment, the same four feeding schedules as in experiment 1 were used with an additional group of larvae (treatment AC) that were fed only on Artemia throughout the rearing period. Similar results were recorded in the two experiments. Larvae in treatments R + A and R4A performed significantly better than those in treatments R8A, ROT and AC. This was particularly evident when examining the proportion of zoeae which successfully completed metamorphosis to megalopa. Poor performance of larvae in treatments AC and ROT implied that rotifers are needed as a first food, but that rotifers alone do not fill the nutritional requirements of S. serrata larvae. Poor performance of larvae in treatment R8A suggested that the diet should be supplemented with Artemia before the end of the zoea 3 stage.     

Rearing dover sole larvae onTisbe andArtemia diets

The suitability of the harpacticoid copepodTisbe holothuriae as a diet for larval and juvenile Dover sole (Solea solea) was assessed by rearing groups of sole for 42 days under a range of dietary regimes. Larval sole, approximately 1 week old, were reared onTisbe, Artemia, or a mixedTisbe-Artemia diet for 13 days. No significant differences in length between sole larvae from any diet were found after this time, but larvae offeredArtemia alone showed a significantly higher frequency of malpigmentation than those offered the other diets. After metamorphosis (day 16), survivors of this experiment were reared for a further 29 days on various diets to give the following dietary sequences:Tisbe-fed larvae, fed onArtemia as juveniles (Tis.-Art.);Artemia-fed larvae, fed onArtemia as juveniles (Art.-Art.);Artemia-fed larvae, fed onTisbe as juveniles (Art.-Tis.) and mixed diet fed larvae, fed on a mixed diet as juveniles (AT-AT). At the end of this period AT-AT and Tis.-Art.-fed juveniles were significantly larger than those on the Art.-Art. dietary regime. Juveniles from the Tis.-Art. dietary regime consumed more prey items than the Art.-Art group. AT-AT juveniles consumed similar amounts of food to Art.-Art. juveniles but were significantly larger after 29 days culture. This was attributed to the presence ofTisbe in their diet. Overall, the best larval and juvenile diet appeared to be a mixed diet throughout the culture period.     

Use of the freshwater rotifer <Emphasis Type="Italic">Brachionus angularis</Emphasis> as the first food for larvae of the Siamese fighting fish <Emphasis Type="Italic">Betta splendens</Emphasis>

Larvae of the Siamese fighting fish Betta splendens were reared on the mass-cultured small freshwater rotifer Brachionus angularis Laos strain (UTAC-Lao), Paramecia sp., and Artemia as live food sources. Larvae fed live food were found to have a significantly high survival rate (97.5–100%) 18 days after hatch (DAH) in comparison to the control unfed larvae, which died by 12 DAH. Rotifer-fed larvae were found to grow faster than paramecia-fed larvae. The fastest growth rate was observed in larvae fed a combination of rotifer and Artemia, with growth in these larvae increasing by 282% by 18 DAH [total length (TL) 11.3 ± 1.2 mm] relative to body measurements taken 3 DAH. The next fastest growth rate was observed in rotifer-fed larvae, with a 158% increase in growth observed by 18 DAH (TL 7.6 ± 0.5 mm). The paramecia-fed larvae were found to grow by only 54.3% (TL 4.6 ± 0.1 mm) during the same period.     

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.     

Iodine‐enriched rotifers and Artemia prevent goitre in Senegalese sole (Solea senegalensis) larvae reared in a recirculation system

To study the effect of dietary supplementation of iodine in Solea senegalensis, larvae were randomly distributed in six tanks. Larvae in three tanks were given rotifers and Artemia enriched with iodine in addition to Rich Advance or Super Selco from 2 days after hatch (DAH) until 31 DAH. Larvae in a second set of three tanks were fed control rotifers and Artemia, enriched only with Rich Advance or Super Selco. Samples were collected at 2, 5, 10, 15 and 31 DAH to determine dry weight, total length, myotome height and thyroid status. Larvae fed the iodine‐enriched diet had significantly higher weight at 31 DAH and higher levels of whole body iodine concentration, compared to control larvae. At 31 DAH, larvae from the control treatment showed typical goitrous thyroid follicles. Thyroid cells of larvae from this treatment appeared columnar or afollicular, with the colloid partly or completely depleted, representative of hyperplasia (goitre). The lower growth rate in fish larvae from the control treatment was possibly a consequence of the hyperplasia, and the iodine enrichment prevented Senegalese sole larvae from developing goitre. This study demonstrates the importance of iodine enrichment of live feed for fish reared in a recirculation system.     

Effects of daphnia (Moina micrura) plus chlorella (Chlorella pyrenoidosa) or microparticle diets on growth and survival of larval loach (Misgurnus anguillicaudatus)

Culture performance beyond metamorphosis of larval loach (Misgurnus anguillicaudatus) was examined in a feeding experiment of the early development stage (20 days after hatch; DAH). Total length, dry weight, length- and weight-specific growth rate (SGR) and survival were monitored in different diet regimes. During 20 days, diet treatments included: microparticle diets (A); live daphnia (Moina micrura) (B); live daphnia plus live chlorella (Chlorella pyrenoidosa) (C); and live daphnia plus microparticle diets (D). Fish survival rates during 20 days were 21.23 ± 4.2% (A), 73.19 ± 2.8% (B), 90.76 ± 3% (C) and 91.46 ± 3.1% (D), respectively. Length- and weight-specific growth rate after 20 DAH (final mean SGR; % day⁻¹) were 5.36 ± 0.44 and 15.75 ± 1.52 (A), 9.29 ± 1.25 and 23.47 ± 2.23 (B), 9.42 ± 1.55 and 24.88 ± 2.9 (C) and 9.55 ± 1.23 and 24.40 ± 2.75 (D), respectively. Fish in treatments B, C and D displayed higher growth rates and were significantly longer and heavier than fish in treatment A by the end of the experiment (Ρ < 0.05). Fish in treatment A had highly significant greater (Ρ < 0.001) mortalities than in treatments B, C and D. There were no significant differences in any growth parameter between fish in treatments B, C and D, but the survivals in treatments C and D (90.76% and 91.46%) were significantly higher than in treatment B (73.19%, Ρ < 0.05). The results demonstrated that enriched prey and co-feeding may serve as a potential feeding strategy for loach larvae, and the form of co-feeding reduces the costs and dependence on live foods to a certain extent. We concluded that larval loach should be reared over metamorphosis using either of the following methods: feed with live daphnia supplemented with microparticle diets or with live chlorella. However, a prolonged rearing period of loach larvae is needed to detect nutritional problems and observe remote effects of co-feeding on weaning in the future.     

Optimal first feed organism for South African mud crab <Emphasis Type="Italic">Scylla serrata</Emphasis> (Forskål) larvae

It is not known whether rotifers or Artemia nauplii are the best first food for South African mud crab Scylla serrata larvae. In order to test this, larvae were fed with five different test diets. These were rotifers for the first 8 days and newly hatched EG^® type Artemia nauplii (San Francisco Bay) from day 6 onwards (treatment R6A); newly hatched EG^® type Artemia nauplii throughout the rearing period (treatment EG); newly hatched Vinh-Chau strain (Vietnam) Artemia nauplii throughout the rearing period (treatment VC); decapsulated cysts of EG^® type Artemia throughout the rearing period (treatment DECAP); or decapsulated cysts supplemented with low densities of Artemia EG type Artemia nauplii (treatment MIX). Two experiments were conducted approximately 1 month apart using larvae from two different female crabs. Although results showed it is possible to rear S. serrata larvae through metamorphosis on Artemia nauplii exclusively, larval performance (development, survival and successful metamorphosis) was enhanced by the inclusion of rotifers as a first feed.No significant difference in performance was recorded between larvae fed on the two strains of Artemia nauplii. Larvae fed on decapsulated cysts in treatments DECAP and MIX performed poorly, but there were indications that decapsulated cysts and other inert diets may have potential as supplements to live food in the rearing of S. serrata larvae.     

Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae: Effects on growth performance and digestive enzyme activities

In this study, the influence of commercial probiotic, Lactobacillus spp., supplementation was investigated on growth parameters and digestive enzyme activities in gilthead sea bream, Sparus aurata, during larval development. All experiments were triplicated and designed in three different administrations of probiotic from 3 days after hatching (DAH) concurrently with starting of exogenous feeding. In the first group, probiotic was added to live food (rotifer and Artemia). In the second group, probiotic was supplemented directly to both live food and water. In the third group, probiotic was added directly to water. Also, no probiotic treatment was maintained in control group. Total bacterial counts among probiotic probiotic-supplemented groups were significantly different from total bacterial counts in controls in water and digestive tract of larvae (p < 0.05). The mean of total bacterial counts in control was approximately 4 × 10⁴-fold increased from the experimental groups in the sea water (p < 0.05). Besides, mean digestive enzyme activities of all probiotics treatment groups were significantly different (p < 0.05) with that of the control. Except probiotic water supplementation group, in all treatments, the specific activities of pancreatic and intestinal enzymes were significantly higher (p < 0.05) in larvae to which probiotic had been supplemented by live food and live food with water. Also, S. aurata larvae that had probiotic administered by live food with water demonstrated significant (p < 0.05) increases in both survival (13–105% higher) and specific growth rate (2–9% higher) as compared to controls. As a result, supplementation of probiotic to directly tank water could not significantly increase growth parameters and digestive enzyme activities and therefore, administration of probiotics by this method would not be effective in terms of husbandry parameters and nutritional condition.     

黑点青鳉的生长和摄食行为发育

针对黑点青鳉实验室养殖成活率低的问题,对其摄食行为发育进行研究,了解其早期阶段的摄食行为发育状况,为人工养殖提供理论依据。通过单摄像机结合镜面成像的方法,对黑点青鳉胚后发育阶段90 d摄食行为的观察与分析,研究了投喂卤虫无节幼体条件下黑点青鳉的生长特征和摄食行为发育。结果显示,幼体体长平均生长率为2.579%/d,体长与日龄间关系为y=3.132+0.383x-0.004x^2+0.00003x^3,R^2=0.98。通过各项摄食行为指标将其幼体生长发育分为3期6个阶段:仔鱼期(0~10 d)分为前仔鱼期(0~4 d)和后仔鱼期(5~10 d),稚鱼期(11~30 d)分为前稚鱼期(11~18 d)、中稚鱼期(19~24 d)和后稚鱼期(25~30 d),幼鱼期(31~65 d)。黑点青鳉幼体出膜4 d后开始投喂初孵卤虫,至10 d所有鱼苗均能捕食。黑点青鳉仔鱼期幼体死亡率较高,但摄食能力稳步提升,至该期末,摄食成功率提升至45%~55%,摄食效率达0.5~0.6个/min;稚鱼期幼体的各项摄食指标均有大幅度提升,对饵料的响应时间不断缩短,摄食速率提高,响应距离也逐渐增长,摄食量增大,摄食成功率已提升至90%~95%;幼鱼期各项摄食指标已接近成鱼,摄食效率达9个/min以上,摄食功能已趋于完善。研究表明,黑点青鳉在仔鱼期对卤虫无节幼体的响应距离短、摄食速率低下,摄食成功率与摄食效率均处于较低水平,随着其自身的发育以及摄食能力的稳定提升,摄食成功率和摄食效率逐步提高,成活率趋于稳定。养殖期间,通过密切注意黑点青鳉的摄食行为变化,改变投喂的数量和频次,使更多的仔鱼尽快过渡到稚鱼期,可能是提高养殖成活率的一条有效途径。     

Improving weaning strategies for Senegalese sole: effects of body weight and digestive capacity

To optimize Senegalese sole‐weaning strategies, three experiments were performed. The first trial tested four weaning strategies with a 10 mg sole. Artemia‐fed sole grew threefold less than fish fed an inert diet. Sudden weaning (abrupt change from Artemia to inert diet) and weaning with co‐feeding produced larger sole than did a late weaning treatment; delayed weaning negatively affected fish growth. In the second experiment, the digestive capacity of early‐weaned 1, 2 and 4 mg sole was investigated. The highest growth was observed in sole weaned at 4 mg. Digestive enzyme profiles suggest that sole have an adaptation period to inert diets, with reduced feed intake. This adaptation period is inversely proportional to post‐larvae weight. The third experiment examined weaning with co‐feeding at different weights (2, 5 and 11 mg). These studies demonstrate that sole of 5–10 mg can be weaned, with high survival rates. On the basis of the digestive enzyme profiles, the early introduction of inert diets in co‐feeding with Artemia seems to affect intestinal processes in smaller postlarvae. This study also suggests that trypsin and alkaline phosphatase may be used as indicators of nutritional status in sole of <5 mg.     

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.     

Prediction of apparent protein digestibility of ingredients and diets by in vitro pH-stat degree of protein hydrolysis with species-specific enzymes for juvenile Pacific white shrimp Litopenaeus vannamei

Aquafeed production faces global issues related to availability of feed ingredients. Feed manufacturers require greater flexibility in order to develop nutritional and cost-effective formulations that take into account nutrient content and availability of ingredients. The search for appropriate ingredients requires detailed screening of their potential nutritional value and variability at the industrial level. In vitro digestion of feedstuffs by enzymes extracted from the target species has been correlated with apparent protein digestibility (APD) in fish and shrimp species. The present study verified the relationship between APD and in vitro degree of protein hydrolysis (DH) with Litopenaeus vannamei hepatopancreas enzymes in several different ingredients (n = 26): blood meals, casein, corn gluten meal, crab meal, distiller's dried grains with solubles, feather meal, fish meals, gelatin, krill meals, poultry by-product meal, soybean meals, squid meals and wheat gluten. The relationship between APD and DH was further verified in diets formulated with these ingredients at 30% inclusion into a reference diet. APD was determined in vivo (30.1 ± 0.5 °C, 32.2 ± 0.4‰) with juvenile L. vannamei (9 to 12 g) after placement of test ingredients into a reference diet (35 g kg^− 1 CP; 8.03 g kg^− 1 lipid; 2.01 kcal g^− 1) with chromic oxide as the inert marker. In vitro DH was assessed in ingredients and diets with standardized hepatopancreas enzymes extracted from pond-reared shrimp. The DH of ingredients was determined under different assay conditions to check for the most suitable in vitro protocol for APD prediction: different batches of enzyme extracts (HPf5 or HPf6), temperatures (25 or 30 °C) and enzyme activity (azocasein): crude protein ratios (4 U: 80 mg CP or 4 U: 40 mg CP). DH was not affected by ingredient proximate composition. APD was significantly correlated to DH in regressions considering either ingredients or diets. The relationships between APD and DH of the ingredients could be suitably adjusted to a Rational Function (y = (a + bx)/(1 + cx + dx²), n = 26. Best in vitro APD predictions were obtained at 25 °C, 4 U: 80 mg CP both for ingredients (R² = 0.86; P = 0.001) and test diets (R² = 0.96; P = 0.007). The regression model including all 26 ingredients generated higher prediction residuals (i.e., predicted APD − determined APD) for corn gluten meal, feather meal, poultry by-product meal and krill flour. The remaining test ingredients presented mean prediction residuals of 3.5 points. A model including only ingredients with APD > 80% showed higher prediction precision (R² = 0.98; P = 0.000004; n = 20) with average residual of 1.8 points. Predictive models including only ingredients from the same origin (e.g., marine-based, R² = 0.98; P = 0.033) also displayed low residuals. Since in vitro techniques have been usually validated through regressions against in vivo APD, the DH predictive capacity may depend on the consistency of the in vivo methodology. Regressions between APD and DH suggested a close relationship between peptide bond breakage by hepatopancreas digestive proteases and the apparent nitrogen assimilation in shrimp, and this may be a useful tool to provide rapid nutritional information.     

The proteolytic digestive activity and growth during ontogeny of Parachromis dovii larvae (Pisces: Cichlidae) using two feeding protocols

The proteolytic digestive activity and growth of Parachromis dovii larvae during the ontogeny were evaluated in a recirculation system using two feeding strategies during a 28-day period. Larvae were reared using two feeding protocols (three replicates each): (A) Artemia nauplii (at satiation), fed from exogenous feeding [8 days after hatching (DAH)] until 15 DAH followed by nauplii substitution by formulated feed (20 % day^?1) until 20 DAH and then formulated feed until 28 DAH; (B) formulated feed (100 % BW daily) from exogenous feeding until 28 DAH. Levels of acid (pepsin type) and alkaline digestive proteases as well as growth and survival of larvae were measured along the feeding period. Survival was high and similar between treatments: 98.9 ± 0.0 for Artemia, 97.3 ± 0.0 % for formulated feed. The specific growth rate for length and weight was higher in larvae fed with Artemia nauplii than in larvae reared with formulated feed: 3.4 ± 0.1 versus 1.8 ± 0.1 % day^?1 for body length (P = 0.009) and 12.2 ± 0.1 versus 6.5 ± 0.3 % day^?1 for body weight (P = 0.002). The acid and alkaline proteolytic activity was detected, in both treatments, from the beginning of the experiment, at 8 DAH. The total enzymatic activity (U larva^?1) for acid and alkaline proteases was higher in larvae reared with Artemia after 12 DAH, whereas the specific enzymatic activity was similar for both enzyme types in the two treatments. The results suggest that P. dovii larvae were capable to digest formulated diets from the beginning of exogenous feeding and that they could be reared with formulated feeds. However, the formulated feed used should be nutritionally improved because of the poor growth obtained in this research.     

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

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

First feeding of freshwater fish larvae with live feed versus compound diets: a meta-analysis

A meta-analysis of published results was used to quantify differences in mortality and growth of freshwater fish larvae when live feed was replaced by compound diets at first feeding. A mean relative risk of 2.4560 (95% confidence interval = 2.0879–2.8891), calculated with 75 observations from 47 studies conducted with 27 freshwater fish species according to a random effects model, indicated that larvae fed on compound diets have a 2.5 times higher chance to die than those fed on live feed. Compared to Artemia nauplii as sole live feed, compound diets were more effective (causing a lower mortality) when replacing zooplankton other than Artemia nauplii. A mean effect size (Hedges’ d) of −3.1813 (95% confidence interval = −3.8099 to −2.5527), calculated (random effects model) from 51 values determined in 33 studies with 21 fish species, represents the size of the negative effect that compound diets would have on growth of larvae. Numerical differences obtained in this study could be use to monitor future development of larval diets.     

Effect of dietary vitamin A on Senegalese sole (Solea senegalensis) skeletogenesis and larval quality

The effects of different levels of vitamin A (VA) in Senegalese sole larval performance and development were evaluated by means of a dietary dose–response experiment using enriched Artemia metanauplii as a carrier of this micronutrient. Larvae were fed from 6 to 27 days post hatch (dph) with enriched Artemia containing graded levels of total VA (1.3, 2.1, 4.5 and 12.9 µg VA mg^− 1 DW). The content of VA in live prey directly affected its accumulation in larvae and early juveniles. Retinyl palmitate accumulated during larval ontogeny, whereas retinol showed the opposite trend, decreasing from hatching until 41 dph and then remaining constant until the end of the study.In metamorphic larvae (10 and 15 dph), VA did not affect the number of thyroid follicles or the intensity of the immunoreactive staining of T₃ and T₄. However, at older stages of development (post-metamorphic larvae: 20, 30, 41 and 48 dph), VA decreased the number of thyroid follicles but increased their mean size and enhanced T₃ and T₄ immunoreactive staining. A dietary excess of VA did not affect either larval performance in terms of growth and survival or the maturation of the digestive system. However, the most remarkable impact of this morphogenetic nutrient was detected during skeletal morphogenesis. Dietary VA accelerated the intramembranous ossification of vertebral centrums, which led to the formation of a supranumerary haemal vertebra and a high incidence of fused and compressed vertebrae in fish fed 2.1, 4.5 and 12.9 mg VA mg^− 1 DW. In addition, VA also affected those structures from vertebrae and caudal fin formed by chondral ossification, leading to defects in their shape and fusions with adjacent skeletal elements. In particular, the caudal fin was the region most affected by the dietary treatments. In order of importance, the bones with more developmental anomalies were the modified neural and haemal spines, epural, hypurals and parahypural. The impact of systemic factors such as thyroidal hormones in skeletogenesis should not be neglected since present results revealed that an excess of dietary VA affected the levels of T₃ and T₄, which might have affected bone formation and remodelling, leading to skeletal deformities.