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In Stung Treng Province north-east Cambodia, WorldFish in partnership with the Fisheries Administration (FiA) and the non-government organisation, Culture and Environment Preservation Association (CEPA), used community science to improve the uptake of small-scale aquaculture (SSA) by communities with limited space and experience of fish culture. The project was funded by the Wetlands Alliance Program and the SSA system, called "WISHPonds" that combines the words "Water and Fish" to reflect the integration of water and fish cultivation with water for storage and vegetable growing. WISH used participatory action research to establish a system of transformative learning in peri-urban households in northeast Cambodia, to assess and evaluate the costs and benefits of establishing SSA in the community. The WISH-Pond system has also been designed to promote the role of gender within the process of SSA development. This paper describes the WISH-Pond system and how research has been used by the community to test and develop aquaculture ponds that meet the needs of households, women in particular. This paper explores the role of gender in community science and in the development and adoption of SSA systems as an alternative livelihoods and contributions to improving management of wetland resources. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the effects of three formulated diets: wheat-gluten-protein-based diets supplemented with Lys- Gly dipeptide ( LG) or free lysine and glycine ( FL), a control diet without lysine supplementation ( C) and commercial starter Bio Oregon ( BO) for on the growth and digestive system morphology of yellow perch. After 48 days of experimental feeding, fish fed LG diet showed the highest body mass. Fish fed LG diet showed the highest number of gastrin/cholecystokinin positive cells and the lowest number of CD3-positive cells. The brush border of anterior intestine was the most Pep T1 immunopositive in fish fed LG diet, the weakest in fish fed C diet. The largest hepatocytes were observed in fish fed BO, while the smallest in those fed FL diet, the difference being statistically significant. Relative hepatocyte cytoplasm volume occupied by lipids was higher in fish fed BO and FL compared to those fed C and LG. Number of proliferating cell nuclear antigen-positive hepatocyte nuclei did not significantly differ among experimental groups. These results indicate that wheat-gluten-protein-based diets supplemented with dipeptide Lys- Gly ( LG) were appropriate for yellow perch. [ABSTRACT FROM AUTHOR]

We have shown previously that diets in which the nitrogen portion is based on synthetic dipeptides (PP) resulted in weight gain by rainbow trout alevins when free amino acid (FAA) based diets did not. However, the protein-based diet used as a control in the previous study resulted in a significantly better performance of fish than peptide- or FAA based diet fed fish. Therefore, the objectives of our study were (i) to test how stomachless fish respond to peptide-based diets, (ii) to evaluate PP and protein-PP mixture diets and (iii) to examine if post-prandial response to FAA concentrations in the fish body can be used as an indicator of the availability of dietary amino acid sources. The first experiment was conducted with a 4-day old Koi carp ( Cyprinus carpio) larvae and included groups fed a casein–gelatin (CG) based diet, a commercial diet and live Artemia nauplii. In the second experiment, fish fed live Artemia for 3 weeks (69 ± 12 mg) were placed in glass aquaria in triplicate per dietary treatment. Four diets provided equivalent amounts of nitrogen in the form of a CG, a PP, a 50% PP and 50% CG (PP50) mix and as FAA. Fish were fed at 1 and 3 h intervals, and the survival and growth were monitored during the second and third weeks of experiments 1 and 2 respectively. Following the completion of feeding, juvenile carp were sampled prior to feeding and 3 h after a meal. Whole body FAA analyses were carried out. We observed marginal suitability of the CG diet for larval Koi carp, as earlier indicated in common carp. Juvenile Koi carp fed the CG diet achieved 236 ± 19 mg, whereas PP50, PP and FAA diet fed fish grew to 140 ± 37, 70 ± 8 and 73 ± 5 mg respectively. Free amino acids in the fish body, and in particular indispensable amino acids (IDAA), were excellent indicators of dietary availability. The present experiment shows that a dietary 1:1 ratio of protein to synthetic PPs results in better growth, survival and whole body IDAA concentrations in relation to FAA- and PP-based diets confirming earlier reports on the use of synthetic PP-based diets, although this diet does not match the performance of complete protein, CG-based diet. [ABSTRACT FROM AUTHOR]

Gender identification of fish species is carried out mainly by examining external morphological characteristics, which in general, it is very complex and not always a reliable approach. Electrophoresis of plasma proteins can be used as an alternative and useful molecular tool for a more precise sex determination. The presence of female specific proteins in the plasma is a starting point for the application of this technique. In this study, reducing discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was applied to analyze plasma proteins of male and female koi carp (Cyprinus carpio haematopterus). Image analyses of electrophoregrams with resolved plasma proteins by SDS-PAGE showed that it is an appropriate technique to discriminate male from female samples. It is based on the presence of apolipoprotein B-100 which can be used as a suitable marker. Further amino acid characterization of apolipoprotein B-100 confirmed that it is a specific protein for female individuals.

Evidence suggests that fish are more tolerant than mammals to imbalanced dietary amino acid profiles. However, the behavioral and physiological responses of fish to individual deficiencies in dietary indispensable amino acids (IDAA) remain unclear. This study examined how stomachless fish respond to diets deficient in limiting IDAA (lysine, methionine, and threonine), using Zebrafish (Danio rerio) as a model. The response to deficient diets was assessed based on; 1) growth performance and feeding efficiency; 2) feed intake; 3) expression of appetite-regulating hormones and nutrient-sensing receptors; and 4) muscle postprandial free amino acid (FAA) levels. There were 6 treatments, each with 3 replicate tanks. A semi-purified diet was formulated for each group. The CG diet was based on casein and gelatin, while the FAA50 diet had 50 % of dietary protein supplied with crystalline amino acids. Both were formulated to contain matching, balanced amino acid profiles. The remaining diets were formulated the same as the FAA50 diet, with minor adjustments to create deficiencies in selected IDAA. The (-) Lys, (-) Met, and (-) Thr diets had lysine, methionine, and threonine withheld from the FAA mix, respectively, and the Def diet was deficient in all three. The juvenile Zebrafish were fed to satiation 3 times daily from 21 to 50 days-post-hatch. Results showed that 50 % replacement of dietary protein with crystalline amino acids significantly reduced growth of juvenile Zebrafish. There were no significant differences in growth between the FAA50 group and groups that received deficient diets. The deficiency of singular IDAA did not induce significant changes in feed intake; however, the combined deficiency in the Def diet caused a significant increase in feed intake. This increased feed intake led to decreased feeding efficiency. A significant decrease in feeding efficiency was also observed in the (-) Lys group. There was an observed upregulation of neuropeptide Y (NPY), an orexigenic hormone, in the Def group. Overall, results from this study suggest stomachless fish increase feed intake when challenged with IDAA-deficient diets, and the regulation of NPY might play a role in this response., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Soybean meal (SBM) has become a common dietary replacement for fish meal (FM) in aquafeed. However, at high inclusions, SBM has been shown to have negative impacts presenting as reduced feed intake and intestinal inflammation. Medicinal plant extracts, namely essential oils, have been used to promote growth performance and immune response. The objective of this study was to investigate the potential therapeutic effects of oregano ( Origanum vulgare ) essential oil (OEO) inclusion on utilization of a high-inclusion SBM diet using zebrafish as a model. Five diets were used in this study: reference-FM-based diet, control-55.7% inclusion SBM diet, and three experimental SBM-based diets OEO1, OEO2, and OEO3 that were supplemented with 1%, 2%, or 3% of oregano oil, respectively. The FM group had overall better growth performance when compared with the other treatment groups; however, the OEO3 mean weight and feed conversion ratio were not significantly different from the FM group ( p > 0.05) and were significantly improved compared with the SBM group ( p < 0.05). Similarly, OEO2 total length was not significantly different from FM ( p > 0.05) but significantly higher than the SBM group ( p < 0.05). Expression of inflammation-related genes did not significantly differ between the OEO groups and the SBM-only group. However, the OEO2 and OEO3 groups displayed improved growth performance compared with the SBM group, suggesting that inclusion of OEO at or above 2% inclusion may help to alleviate common symptoms induced by a high-inclusion SBM diet.

Fish protein hydrolysates used in larval diets have been prepared from a variety of fish species, with different enzymes used to hydrolyze the protein. This study's objectives were to determine the effect of the dietary inclusion of fish muscle hydrolysates obtained from species-specific muscle/enzymes-versus hydrolysates produced from muscle/enzymes of a different species-on the growth performance, survival, skeletal development, intestinal peptide uptake, and muscle-free amino acid (FAA) composition of larval Walleye ( Sander vitreus ). Eight protein products were obtained for this study, comprising an unhydrolyzed and hydrolyzed product from each combination of muscle/enzymes from Walleye and Nile tilapia ( Oreochromis niloticus ). Four diets were produced, and the dietary protein was provided in a 50/50 ratio of unhydrolyzed and hydrolyzed protein from the respective muscle/enzyme combination. Four groups were fed one of the corresponding formulated diets, and two groups of larvae, fed a commercial starter diet and Artemia , respectively, served as reference groups. Larval Walleye fed the diet containing protein produced with the species-specific muscle and enzymes had a significantly higher weight after the study-30% higher than any other group. A significant interaction effect between muscle and enzyme sources on the growth of Walleye larvae was observed. The species-specific combination also led to a significant increase in postprandial FAA and indispensable amino acid concentrations in muscle. No significant differences were observed between the hydrolysate-fed groups in survival, deformity occurrence, or peptide uptake. Each hydrolysate-based diet significantly reduced skeletal deformities and survival compared to the commercial diet. The results of this study suggest that species-specific muscles and enzymes produce a more optimal dietary protein source for larval fish than non-species-specific products. Further research should focus on improving the physical properties of the formulated diets to reduce possible leaching of hydrolyzed protein and improve the survival of fish larvae.

Many organisms exhibit social behaviors and are part of some scheme of social structure. Zebrafish are highly social, shoaling fish and therefore, social isolation may have notable impacts on their physiology and behavior. The objective of this study was to evaluate the effects of social isolation on feed intake, monoaminergic system related gene expression, and intestinal health of juvenile zebrafish fed a high-inclusion soybean meal based diet. At 20 days post-fertilization zebrafish were randomly assigned to chronic isolation (1 fish per 1.5 L tank) or social housing (6 fish per 9 L tank) with 18 tanks per treatment group (n = 18). Dividers were placed between all tanks to prevent visual cues between fish. Zebrafish were fed a commercial fishmeal based diet until 35 days post-fertilization and then fed the experimental high-inclusion soybean meal based diet until 50 days post-fertilization. At the end of the experiment (51 days post-fertilization), the mean total length, weight, and weight gain were not significantly different between treatment groups. Feed intake and feed conversion ratio were significantly higher in chronic isolation fish than in social housing fish. Expression of monoaminergic and appetite-related genes were not significantly different between groups. The chronic isolation group showed higher expression of the inflammatory gene il-1b, however, average intestinal villi width was significantly smaller and average length-to-width ratio was significantly higher in chronic isolation fish, suggesting morphological signs of inflammation were not present at the time of sampling. These results indicate that chronic isolation positively affects feed intake of juvenile zebrafish and suggest that isolation may be useful in promoting feed intake of less-palatable diets such as those based on soybean meal., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Dissinger et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This study's objectives were to determine the effect of Largemouth Bass (LMB) muscle hydrolysates obtained using same-species digestive enzymes and the degree of LMB muscle hydrolysis when included in the first feeds of growth performance and survival, skeletal development, intestinal peptide uptake, and muscle-free amino acid composition of larval LMB. LMB muscle was mixed with digestive enzymes from adult LMB, and hydrolyzed for 1.5, 3, and 6 h, respectively. Five diets were produced, the intact diet containing non-hydrolyzed muscle and four diets with 37% muscle hydrolysate inclusion. Those diets were characterized by their level of each hydrolysate (presented as a ratio of 1.5, 3, and 6 Ts hydrolysates): 1:1:1, 1:3:6, 1:3:1, 6:3:1 for diets A, B, C, and D, respectively. To account for gut development, one group of larval LMB was fed a weekly series of diets B, C, and D to provide an increasing molecular weight profile throughout development. This group was compared against others that received either; (1) diets D, C, and B; (2) diet A; or (3) intact diet. The initial inclusion of the hydrolysates significantly improved the total length of the larval LMB; however, neither the hydrolysate inclusion nor the series of dietary molecular weight profiles improved the overall growth of larval LMB. The inclusion of hydrolysates significantly decreased the occurrence of skeletal deformities. The degree of hydrolysis did not have a significant effect on the parameters measured, except for intestinal peptide uptake, which was increased in the group that received the most hydrolyzed diet at the final time of sampling. The lack of overall growth improvement suggests that while the hydrolysates improve the initial growth performance, further research is necessary to determine the optimal molecular weight profile, hydrolysate inclusion level, and physical properties of feeds for larval LMB.

Live food is necessary for the proper development of zebrafish larvae, providing nutrition in a form that is easily digestible and available to the larvae. Live food is commonly enriched to increase the dietary content of certain nutrients. However, little research has been done on protein-based enrichments, especially those of plant origin. This study sought to examine how different quality protein enrichments affected the composition of live food as well as growth and digestive tract development of larval zebrafish, Danio rerio . Larval zebrafish were fed from 3 to 22 days posthatch (dph) with one of six live food (rotifers Brachionus plicatilis and Artemia spp.) treatments: (1) live food with no enrichment (starved; control); (2) live food enriched with commercially used Spirulina spp. algae; (3) live food enriched with soybean meal (SBM); (4) live feed enriched with soy protein concentrate (SPC); (5) live feed enriched with a fishmeal hydrolysate; and (6) live feed enriched with intact fishmeal (FM). Proximate composition of live food was significantly affected by enrichment, in particular, protein content of rotifers was significantly increased by enrichment with SBM. Zebrafish fed SBM-enriched live food showed longer total body length than all other groups, except SPC. Zebrafish in the SBM group also showed increased gene expression of chymotrypsin in the intestine, possibly indicating improved intestinal development and extracellular digestion, which likely contributed to improved growth. Conversely, zebrafish fed hydrolysate-enriched live food showed reduced gene expression of alkaline phosphatase, possibly indicating a less developed intestinal tract, correlating with reduced growth compared to SBM group. Overall, plant protein was shown to be a promising source of live food enrichment for improving larval zebrafish growth.

Plant protein (PP) utilization in fish is limited due to lower digestibility compared to fishmeal (FM) and the presence of antinutritional factors. Its utilization can be improved by nutritional programming (NP), a method wherein a fish is provided a nutritional stimulus early in life which can alter their physiology. NP has been shown to be effective but methods of applying NP are varied and have been applied at different stages of development with different outcomes. To find the optimal timeframe to perform NP in fish early stages Largemouth bass (Micropterus salmoides, Lacepède) were nutritionally programmed at three different ages in early development. In this study bass were programmed with: (1) live food enriched with soybean meal (SBM) from 6 to 15 days post-hatch (dph) (NPL), (2) SBM-based formulated diet from 16 to 25 dph (NPD1) and (3) formulated SBM-diet from 26 to 35 dph (NPD2). After programming, each group was fed FM-diet before being refed SBM-diet from 100 to 172 dph. A positive control (PC) was fed FM-diet throughout. Final average body weight of PC was significantly higher than NPD1 and NPD2 but did not significantly differ from NPL. Overall NPL showed much improved growth and utilization of PP compared to NPD1 and was similar to growth achieved by PC. This study showed an optimum window of time exists wherein NP of Largemouth bass yields the best impact on growth in the larval stage and later in life when fed SBM-diet. Programming should be performed right after mouth opening using enriched live food and can result in growth similar to non-programmed fish fed FM-based diet. Programming effects similar to that of the live food approach can be achieved with formulated diet, however it is crucial that Largemouth bass are of a proper age and sufficiently developed when programmed with dry food or severe impacts on growth can occur., (© 2022 Wiley-VCH GmbH.)

Nutritional programming (NP) is a concept in which early nutritional events alter the physiology of an animal and its response to different dietary regimes later in life. The objective of this study was to determine if NP via broodstock with dietary plant protein (PP) has any effect on the gut microbiome of the progeny fish and whether this modified gut microbiome leads to better utilization of PP diet. The experiment consisted of four different treatments as follows: (1) progeny that received FM diet obtained from fishmeal (FM)-fed broodstock (FMBS-FM, +control); (2) progeny that received PP diet obtained from FM-fed parents (FMBS-PP); (3) progeny that received PP diet obtained from "nutritionally programmed" parents (PPBS-PP; -control); and (4) progeny that received FM diet obtained from "nutritionally programmed" parents (PPBS-FM). Zebrafish was used as a model species. This study found that parental programming seems to have some positive effect on dietary PP utilization in progeny. However, the influence of NP with PP through broodstock on gut microbiota of the offspring fish was not detected.

Manipulating feeding rate and protein quality may improve growth and feeding efficiency of cultured species. However, whether feeding rate, protein quality, or their interaction has a greater effect on growth and feeding efficiency response variables is unknown. To determine whether feeding rate and protein quality individually or interactively affect growth and feeding efficiency, juvenile Zebrafish ( Danio rerio ) were either offered nutritionally similar diet consisting of either menhaden fishmeal protein or a 100% replacement of fishmeal with soybean meal-based protein restrictively or to satiation. Total length, weight, feed intake, and feed conversion ratio (FCR) were measured throughout the duration of the study. Protein quality and feeding rate individually and interactively affected feed intake and FCR: Zebrafish offered feed to satiation had higher growth and FCR than those fed restrictively, and Zebrafish fed soybean meal-based diet showed lower growth and higher FCR and feed intake compared to those fed fishmeal-based diet, although magnitude of response depended on feeding rate. These findings likely indicate lower digestibility of soybean meal or the presence of antinutritional factors in soybean meal that led to impaired nutrient absorption of fish offered soybean meal-based diet. Differences in measured response variables between protein qualities and feeding rates highlight the importance of determining interactive effects in nutritional studies.

The present study utilized digestives tracts from adult largemouth bass (LMB) to hydrolyze Bighead carp muscle and obtain an optimal profile of muscle protein hydrolysates that would be easily assimilated within the primitive digestive tract of larval LMB. Specifically, muscle protein source was digested for the larva using the fully developed digestive system of the same species. The objectives of this study were: 1) to develop an optimal in vitro methodology for carp muscle hydrolysis using LMB endogenous digestive enzymes, and 2) to evaluate the effect of dietary inclusion of the carp muscle protein hydrolysate on LMB growth, survival, occurrence of skeletal deformities, and whole-body free amino acid composition. The study found that the in vitro hydrolysis method using carp intact muscle and LMB digestive tracts incubated at both acid and alkaline pH (to mimic digestive process of LMB) yielded a wide range of low molecular weight fractions (peptides), as opposed to the non-hydrolyzed muscle protein or muscle treated only with acid pH or alkaline pH without enzymes from LMB digestive tracts, which were comprised of large molecular weight fractions (polypeptides above 150 kDa). Overall, the dietary inclusion of the carp muscle hydrolysate improved growth performance of larval LMB in terms of final average weight, weight gain, DGC, SGR, and body length after 21 days of feeding compared to fish that received the diet based on non-hydrolyzed carp muscle. The study also found that hydrolysate-based feed significantly reduced skeletal deformities. The positive growth performance presented by fish in the hydrolysate-fed group possibly resulted from matching the specific requirements of the larvae with respect to their digestive organ development, levels of digestive enzymes present in the gut, and nutritional requirements., Competing Interests: The authors have declared that no competing interests exist.

The objective of the present study was to compare skeletal muscle proteomic profiles, histochemical characteristics, and expression levels of myogenic regulatory factors (MRFs) between fast- versus slow-growing yellow perch Perca flavescens and identify the proteins/peptides that might play a crucial role in the muscle growth dynamic. Yellow perch were nursed in ponds for 6 weeks from larval stage and cultured in two meter diameter tanks thereafter. The fingerlings were graded to select the top 10% and bottom 10% fish which represented fast- and slow-growing groups (31 yellow perch per each group). Our statistical analyses showed 18 proteins that had different staining intensities between fast- and slow-growing yellow perch. From those proteins 10 showed higher expression in slow-growers, and 8 demonstrated higher expression in fast-growers. Fast-growing yellow perch with a greater body weight was influenced by both the muscle fiber hypertrophy and mosaic hyperplasia compared to slow-growing fish. These hyperplastic and hypertrophic growth in fast-grower were associated with not only metabolic enzymes, including creatine kinase, glycogen phosphorylase, and aldolase, but also myoD and myogenin as MRFs. Overall, the results of the present study contribute to the identification of different expression patterns of gene products in fast- and slow-growing fish associated with their muscle growth., (© 2021. The Author(s).)

Soybean meal (SBM) inclusion in aquaculture diets has been found to negatively affect growth and induce intestinal inflammation in fish. The objective of this study was to determine the effect of health-promoting dipeptide supplementation into SBM-based feeds on growth performance, intestinal health, and muscle free amino acid composition, an indicator of dietary amino acid availability, in a zebrafish model. There were five treatment groups in this study. The first group ((+) Control) received a fishmeal-based diet. The second group ((-) Control) received SBM-based diet. The last three groups (Ala-Glu, Car, and Ans) were fed SBM-based diets, supplemented with alanyl-glutamine, carnosine, and anserine respectively. The Ala-Glu and Car groups experienced a significantly higher weight gain than the (-) Control group, weighing 35.38% and 33.96% more, respectively at the conclusion of the study. There were no significant differences in gene expression among the groups, but Ala-Glu had the highest expression of both nutrient absorption genes measured, PepT1 and fabp2. Ala-Glu had significantly longer intestinal villi, and a significantly higher villus length-to-width ratio than the (-) Control group. The Car group had a significantly higher post-prandial tissue concentration of lysine, compared to the (-) Control group. The increase in villus surface area and expression of nutrient absorption genes represent an improvement in intestinal absorptive capacity in the Ala-Glu group. The results from this study provide support for the use of alanyl-glutamine and carnosine supplementation as a means of improving growth performance of zebrafish fed with a high level SBM-based diet., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Nutritional Programming (NP) has been studied as a means of improving dietary plant protein (PP) utilization in different fish species. This study investigated the use of enriched live feed as a vehicle for NP in larval fish. The objective of this study was to determine the effect of NP induced during the larval stage via PP-enriched live feed on: (1) growth performance; (2) expression of genes associated with inflammation and any morphological changes in the intestine; and (3) muscle free amino acid composition in largemouth bass (Micropterus salmoides) during its later life stages. Two diets were used in this study, a fish meal (FM)-based diet, and a soybean mean (SBM)-based diet, serving as the PP diet. There were 4 groups in this study. The two control groups, ( +) Control and (-) Control, were not programmed and received the FM-diet and SBM-diet, respectively throughout the whole trial after the live feed stage (27-122 days post hatch (dph). The next group, programmed, was programmed with SBM-enriched Artemia nauplii during the live feed stage (4-26 dph) and challenged with the SBM-diet during the final stage of the study (79-122 dph). The final group, non-programmed, did not receive any programming and, was challenged with the SBM-diet during the final stage of the study. The programmed group experienced a significantly higher (%) weight gain during the PP-Challenge than the non-programmed group. In addition, the live feed programming resulted in significantly longer distal villi, and a higher villi length to width ratio, compared to the non-programmed group. No significant effects on free amino acid composition and gene expression were observed between the programmed and non-programmed group, except for an increased post-prandial concentration of free proline in the programmed group. The results of this study support use of live feed as a vehicle for nutritional programming and improving the growth performance of largemouth bass fed with a SBM-based diet.

Nutritional programming (NP) is considered a promising approach that can counteract the negative effects of dietary plant protein (PP) by introducing PP to fish in the early developmental stages. Therefore the objective of our study was to assess the effect of NP on PP utilization and the gut microbiome in zebrafish Danio rerio . The study included four treatment groups: (1) a positive control group that received a fishmeal (FM) diet throughout the entire trial (+ control); (2) a negative control group that received PP diet throughout the entire trial (- control); (3) an NP group that received dietary PP during the larval stage followed by FM-based diet during the juvenile stage and PP diet again during a PP challenge in the grow-out phase (NP-PP); and (4) an FM-group that received FM-based diet during the larval and juvenile stages and was challenged with a PP diet during the grow-out phase (NP-FM). During the PP challenge, the NP-PP group achieved the highest weight gain compared to the (-) control and NP-FM groups. The relative abundance of certain phyla such as Chloroflexi, Planctomycetes, and Bacteroidetes presented higher values in some groups at early juvenile stage. The fish gut microbiome also presented differences throughout the study.

Nutritional Programming (NP) has been studied as a means of mitigating the negative effects of dietary plant protein (PP), but the optimal timing and mechanism behind NP are still unknown. The objectives of this study were: 1) To determine whether zebrafish (Danio rerio) can be programmed to soybean meal (SBM) through early feeding and broodstock exposure to improve SBM utilization; 2) To determine if NP in zebrafish affects expression of genes associated with intestinal nutrient uptake; 3) To determine if early stage NP and/or broodstock affects gene expression associated with intestinal inflammation or any morphological changes in the intestinal tract that might improve dietary SBM utilization. Two broodstocks were used to form the six experimental groups. One broodstock group received fishmeal (FM) diet (FMBS), while the other was fed ("programmed with") SBM diet (PPBS). The first ((+) Control) and the second group ((-) Control) received FM and SBM diet for the entire study, respectively, and were progeny of FMBS. The last four groups consisted of a non-programmed (FMBS-X-PP and PPBS-X-PP) and a programmed group (FMBS-NP-PP and PPBS-NP-PP) from each of the broodstocks. The programming occurred through feeding with SBM diet during 13-23 dph. The non-control groups underwent a PP-Challenge, receiving SBM diet during 36-60 dph. During the PP-Challenge, both PPBS groups experienced significantly lower weight gains than the (+) Control group. NP in early life stages significantly increased the expression of PepT1 in PPBS-NP-PP, compared to PPBS-X-PP. NP also tended to increase the expression of fabp2 in the programmed vs. non-programmed groups of both broodstocks. The highest distal villus length-to-width ratio was observed in the dual-programmed group, suggesting an increase in surface area for nutrient absorption within the intestine. The results of this study suggest that NP during early life stages may increase intestinal absorption of nutrients from PP-based feeds., Competing Interests: The authors have declared that no competing interests exist.

Nutritional Programming (NP) has been shown to counteract the negative effects of dietary plant protein (PP) by introducing PP at an early age towards enhancement of PP utilization during later life stages. This study explored the effect of NP and its induction time on growth, expression of appetite-stimulating hormones, and any morphological changes in the gut possibly responsible for improved dietary PP utilization. At 3 days post-hatch (dph) zebrafish were distributed into 12 (3 L) tanks, 100 larvae per tank. This study included four groups: 1) The control (NP-FM) group received fishmeal (FM)-based diet from 13-36 dph and was challenged with PP-based diet during 36-66 dph; 2) The NP-PP group received NP with dietary PP in larval stage via live food enrichment during 3-13 dph followed by FM diet during 13-36 dph and PP diet during 36-66 dph; 3) The T-NP group received NP between 13-23 dph through PP diet followed by FM diet during 23-36 dph and PP diet during 36-66 dph; and 4) The PP group received PP diet from 13-66 dph. During the PP challenge the T-NP group achieved the highest weight gain compared to control and PP. Ghrelin expression in the brain was higher in T-NP compared to NP-FM and NP-PP, while in the gut it was reduced in both NP-PP and T-NP groups. Cholecystokinin expression showed an opposite trend to ghrelin. The brain neuropeptide Y expression was lower in NP-PP compared to PP but not different with NP-FM and T-NP groups. The highest villus length to width ratio in the middle intestine was found in T-NP compared to all other groups. The study suggests that NP induced during juvenile stages improves zebrafish growth and affects digestive hormone regulation and morphology of the intestinal lining-possible mechanisms behind the improved PP utilization in pre-adult zebrafish stages., Competing Interests: The authors have declared that no competing interests exist.

Achieving Sustainable Development Goal 2 of zero hunger and malnutrition by 2030 will require dietary shifts that include increasing the consumption of nutrient dense foods by populations in low- and middle-income countries. Animal source foods are known to be rich in a number of highly bioavailable nutrients that otherwise are not often consumed in the staple-food based diets of poorer populations throughout the world. Fish is the dominant animal source food in many low- and middle-income countries in the global south and is available from both fisheries and aquaculture. Consumers often perceive that wild caught fish have higher nutritional value than fish produced through aquaculture, and this may be true for some nutrients, for example omega-3 fatty acid content. However, there is potential to modify the nutritional value of farmed fish through feeds and through production systems, illustrated by the common practice of supplementing omega-3 fatty acids in fish diets to optimize their fatty acid profile. This manuscript reviews the evidence related to fish feeds and the nutritional composition of fish with respect to a number of nutrients of interest to human health, including iron, zinc, vitamins A and D, selenium, calcium, and omega-3 fatty acids, with low- and middle-income country populations in mind. In general, we find that the research on fortification of fish diet particularly with vitamins and minerals has not been directed toward human health but rather toward improvement of fish growth and health performance. We were unable to identify any studies directly exploring the impact of fish feed modification on the health of human consumers of fish, but as nutrition and health rises in the development agenda and consumer attention, the topic requires more urgent attention in future feed formulations.

Hypoxia can affect various fish populations, including yellow perch Perca flavescens, which is an economically and ecologically important species in Lake Erie, a freshwater system that often experiences hypoxia in the hypolimnetic part of the lake. Fish, similarly to mammals, possess molecular oxygen sensor-hypoxia-inducible factor-1 (HIF-1), a transcription factor that can affect expression of many downstream genes related to animal growth and locomotion, protein synthesis, as well as ATP and amino acid metabolism. HIF-1 is a heterodimer, which consists of two subunits: oxygen-sensitive and oxygen-insensitive subunits, α and β, respectively. In this study, we report first on the molecular cloning and sequencing of P. flavescens HIF-1α. The full-length complementary DNA (cDNA) was isolated and submitted to the GenBank with accession number KT783483. It consists of 3529 base pairs (bp) carrying a single open-reading frame that encompasses 2250 bp of the coding region, 247 bp of the 5' untranslated region (UTR), and 1032 bp of the 3' UTR. The "de novo" prediction of the 3D structure of HIF-1α protein, which consists of 749 amino acids, is presented, too. We then utilized One-Step Taqman® real-time RT-PCR technology to monitor changes in HIF-1α messenger RNA (mRNA) copies in response to chronic hypoxic stress. An experiment was conducted using 14-day post-swim-up stage yellow perch larvae with uninflated swim bladders. This experiment included three treatment groups: hypoxia, mid-hypoxia, and normoxia, in four replicates (four tanks per treatment) with the following dissolved oxygen levels: 3, 4, and >7 mg O 2 /L, respectively. At the end (2 weeks) and in the middle (1 week) of the experiment, fish from each tank were sampled for body measurements and molecular biology analysis. The results showed no differences in survival (∼90%) between treatment groups. Oxygen concentration was lowered to 3.02 ± 0.15 (mean ± SE) mg O 2 /L with no adverse effect on fish survival. The highest growth rate was observed in the normoxic group. A similar trend was observed with fish body length. The growth rate of fish declined with decreasing water-dissolved oxygen. The number of HIF-1α mRNA copies was not significantly different between hypoxic, mid-hypoxic, and normoxic conditions, and this was true for fish obtained in the middle and at the end of the experiment. Graphical abstract.

Studies on larval rearing of silver bream (Vimba vimba), a migratory cyprinid fish have addressed on limited scale larval and juvenile rearing using commercial and semipurified diets along with live feeds, such as brine shrimp Artemia nauplii. The objectives of the present study were (1) to determine whether experimental, protein-, peptide-, free amino acid-based diets are adequate for larval silver bream, a stomachless fish, (2) to evaluate whether commercial and purified diets are comparable as the first/exclusive feed for growth and survival of silver bream, and (3) to examine whether free amino acid concentrations in fish body are potential indicators of availability of amino acid sources. We report here the differences in diets acceptance, fish growth and diet utilization in silver bream in comparison to other cyprinid fishes. We specifically address the response in free amino acids in the body to dietary treatments. Experimental diets included: a commercial Aglo Norse feed, casein-gelatin based diet (CG), free amino acid mixture diet (FAA), dipeptide (PP), dipeptide-protein (PP50) based diet, and dipeptide diet without arginine (NoArg). In addition, live Artemia were offered to 3 groups and "fasting" control treatment was included during 3 week long trial. Fish offered Artemia overperformed those offered formulated diets both in terms of mass (80.7+/-26.3 mg) and survival (97.2%). We also indicate that commercial and purified diets are comparable as the first/exclusive feed for growth and survival of silver bream. Our experiment also showed that the whole body free amino acid concentrations of 9 indispensable amino acids (IDAA) out of 10 (His, Thr, Arg, Val, Met, Ile, Leu, Trp, Lys) in the PP50 group was the highest among 7 diet treatments and the totalfree amino acid concentration, total dispensable amino acids (DAA) and total IDAA of the PP50 diet fed fish showed the same trend. This may indicate that diets based on 50% of dipeptides and 50% of protein are adequate for silver bream larvae as they result in similar growth, survival and whole body IDAA concentrations as in fish fed CG (protein-based) diet fed fish.