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Zebrafish is one of the most used vertebrate model organisms in molecular and developmental biology, recently gaining popularity also in medical research. However, very little work has been done to assess zebrafish as a model species in nutritional studies in aquaculture in order to utilize the methodological toolbox that this species represents. As a starting point to acquire some baseline data for further nutritional studies, growth of a population of zebrafish was followed for 15 weeks. Furthermore, whole body proteome was screened during development by means of bi-dimensional gel electrophoresis and mass spectrometry. Fish were reared under best practice laboratory conditions from hatching until 103 days post-fertilization (dpf) and regularly fed ad libitum with Artemia nauplii from 12 dpf. A growth burst occurred within 9-51 dpf, reaching a plateau after 65 dpf. Fork length and body weight were significantly lower in males than in females from 58 dpf onwards. Proteomics analysis showed 28 spot proteins differently expressed through development and according to sex. Of these proteins, 20 were successfully identified revealing proteins involved in energy production, muscle development, eye lens differentiation, and sexual maturation. In summary, zebrafish exhibited a rapid growth until approximately 50 dpf, when most individuals started to allocate part of the dietary energy intake for sexual maturation. However, proteomic analysis revealed that some individuals reached sexual maturity earlier and already from 30 dpf onwards. Thus, in order to design nutritional studies with zebrafish fed Artemia nauplii, it is recommended to select a period between 20 and 40 dpf, when fish allocate most of the ingested energy for non-reproductive growth purposes. [ABSTRACT FROM AUTHOR]

Ontogenetic changes in the capacity of Senegalese sole (Solea senegalensis Kaup, 1858) larvae to digest and metabolise Artemia protein and amino acids (AA) were studied using 12, 22 and 35 days after hatching (DAH) larvae that were fed Artemia metanauplii radiolabelled with a [U-14C] protein hydrolysate. About 82% and 18% of the label was incorporated into the Artemia trichloroacetic acid (TCA) precipitate (mostly protein) and soluble (mainly free AA) fractions, respectively. The digestibility of Artemia was high at all tested ages, with label absorption varying between 77% and 83% at 24 h after feeding (HAF). A rapid digestion, absorption and catabolism of Artemia AA were noted, with most of the absorption into the body occurring during the first 3 HAF. Traces of label were already found in the metabolic-CO2 trap at 1 HAF. Furthermore, label was largely and almost immediately incorporated into the TCA precipitate fraction (mostly protein) of gut and body tissues. Slight differences were noted in diet utilization between larvae at different ages. At 12 DAH larvae had a lower catabolism and evacuation of the label, as well as the highest accumulation in the gut. However, except for the amount of catabolised label, the results were not significantly different from those for 35 DAH larvae. Moreover, no significant differences were found in the amount of label incorporated into the body, although it seemed to be higher in 12 DAH larvae. Taken together, these results reveal a higher absorption and a significantly higher retention of the absorbed label in 12 DAH larvae. In addition, 12 DAH larvae appeared to have a slower absorption of the label, which, in continuously feeding larvae, might result in overall lower food absorption efficiency. Therefore, it seems that young larvae have the ability to compensate for a possible lower digestive capacity with higher body retention of absorbed AA. This study confirms that sole larvae, even young stages, have a high capacity for digesting live preys. [ABSTRACT FROM AUTHOR]

With the aim of improving the formulation of diets for early stages of marine fish, post-larval Hippoglossus hippoglossus (56 days after first feeding; 23 mg dry weight) were tube-fed liquid diets of radiolabelled L-(14C) free amino acids (FAA diet), peptides (PEPT diet) and protein (PROT diet), in order to compare the kinetics of transfer from gut into the liver and carcass. The test diets were prepared using equal concentrations of amino acids (AA) and fed as a single pulse (0.3 µl, 2.2 µg AA). The PROT diet consisted of methylated-14C-labelled (mainly lysine) bovine serum albumin (BSA), the PEPT diet was the PROT diet digested with trypsin and the FAA diet consisted of an amino acid mixture based on BSA composition with added 14C-lysine. The common labelling with 14C-lysine permitted a comparison of the three diets. Based on dissection and distribution of the tracer in each compartment, the FAA diet was almost completely absorbed 1 h after the tube-feeding; PEPT and PROT diets required more time before they were processed by the digestive tract. The FAA diet was absorbed about eight times as fast as protein and about six times as fast as peptides from the gut. A low retention of label in the liver for all diets at all sampling times suggests that the liver does not store the AA at these absorption rates. The FAA diet had a higher assimilation efficiency (about 90%) than the PEPT diet (about 12%) and PROT diet (about 32%). FAAs, thus, appear to be superior to protein and peptides as a dietary source of amino acids in post-larval Atlantic halibut. [ABSTRACT FROM AUTHOR]

To specifically study the functional ontogeny of the digestive tract, larvae of Japanese flounder at various developmental stages were injected with liquid solutions using tube feeding in vivo. The survival in the stages tested was on average 93%. The injected solutions were almost completely transferred from the presumptive stomach, or stomach to the midgut within 10 min of injection. The passage to the hindgut in some cases started 10 min after injection and over 90% of the solution had passed from the midgut to the hindgut after 1 h. In most cases the hindgut seemed to be completely empty after 3 h. Two different mixtures of pH indicators with sensitivities in the alkaline (7.5 to 9) and acid (4 to 6) ranges respectively were used for assessment of pH in the various gut segments. The pH of the stomach remained alkaline during the larval period, but had fallen close to 4 during late metamorphosis, an indication of active HCl secretion and progressive stomach differentiation. In mid and late metamorphosing fish a rapid colour change in the pH indicator was observed once it had passed the pyloric sphincter. This demonstrates that there was also active secretion of alkaline fluid, most likely HCO3 −, from the pancreas into the pylorus lumen. A single injection of liquid solution of 14C-FAA showed that assimilation of FAA was high in all stages tested (79.5 ± 7.1%; SD; n=91). The presently reported data for Japanese flounder support earlier studies that FAA are absorbed with a high efficiency in the early stages of marine fish. [ABSTRACT FROM AUTHOR]

Free amino acids (FAA) and protein have been measured in whole laboratory-readed halibut larvae and on dissected individuals separated into yolk and body compartments. At hatching both FAA and protein are mainly located in the yolk compartment. During the first 12 d of the yolk-sac stage more than 70% of the FAA pool disappeared from the yolk without any significant changes in the yolk protein pool. This suggests different uptake mechanisms for FAA and protein from the yolk, and a sequential utilisation of the endogeneous reservoirs of free and protein amino acids in Atlantic halibut larvae. The data suggest that in the early yolk-sac stage FAA enter the embryo from the yolk and are utilised both for energy and protein synthesis. Later on when the free pool cannot fulfil the nutritional requirements, additional amino acids are recruited from yolk protein. Of the total amino acids (free + protein amino acids) present at hatching ca. 60% will be used as precursors for body protein synthesis while the remaining 40% are used as fuel in the larval energy metabolism. [ABSTRACT FROM AUTHOR]

Comments on the article "Assimilation of Dietary Free Amino Acids, Peptides and Protein in Post-Larval Atlantic Halibut," published in previous issue. Study conducted on methylated bovine serum albumin (BSA) using the protein and peptide test solutions. Demonstration of the study in vitro hydrolysis rates of methylated BSA by two mammalian cytosolic proteases; Verification of the results for assimilation of protein and peptides with the use of non-methylated compounds.