Microplastic Contamination in Aquafeed Ingredients Used as Protein and Carbohydrate Sources.

The current study aimed to evaluate the occurrence of microplastics in feed ingredients commonly used as protein and carbohydrate (energy) sources to understand and mitigate microplastic contamination. Microplastics average was 1.27, 0.69, 2.85, 0.55, 0.07, and 0.17 particle g- 1 in fishmeal, soybean meal, poultry by-products, rice bran, wheat bran, and wheat flour, respectively. Notably, poultry by-products demonstrated significantly higher microplastic levels than other ingredients (p < 0.05). The dominant microplastic shape was microfibers, with prevalent sizes ranging from 500 to 1000 µm. We estimated that packaging materials are a significant source of pollution due to the high presence of polypropylene and other polymers. Top aquaculture species with the greatest microplastic exposure risk include the Catla catla, Hypophthalmichthys nobilis, and Oreochromis niloticus. This research extends our knowledge of microplastic pathways, contributes to improving aquafeed quality, and provides the basis for determining the risk of microplastic exposure in aquafeed.

Exploring mercury detoxification in fish: The role of selenium from tuna byproduct diets for sustainable aquaculture.

Exposure to mercury (Hg) through fish consumption poses significant environmental and public health risks, given its status as one of the top ten hazardous chemicals. Aquaculture is expanding, driving a surge in demand for sustainable aquafeeds. Tuna byproducts, which are rich in protein, offer potential for aquafeed production, yet their use is challenged by the high content of heavy metals, particularly Hg. However, these byproducts also contain elevated levels of selenium (Se), which may counteract Hg adverse effects. This study examines the fate of dietary Hg and Se in an aquaculture model fish. Biomolecular speciation analyses through hyphenated analytical approaches were conducted on the water-soluble protein fraction of key organs of juvenile rainbow trout (Oncorhynchus mykiss) exposed to various combinations of Hg and Se species, including diets containing tuna byproducts, over a six-month period. The findings shed light on the dynamics of Hg and Se compounds in fish revealing potential Hg detoxification mechanisms through complexation with Hg-biomolecules, such as cysteine, glutathione, and metallothionein. Furthermore, the trophic transfer of selenoneine is demonstrated, revealing novel opportunities for sustainable aquafeed production. Understanding the interactions between Hg and Se in aquaculture systems is crucial for optimizing feed formulations and mitigating environmental risks. This research contributes to the broader goal of advancing sustainable practices in aquaculture while addressing food security challenges.

Transfer and bioaccumulation of chemical and biological contaminants in the marine polychaete Hediste diversicolor (OF müller 1776) when reared on salmon aquaculture sludge.

Side streams from aquaculture production such as fish sludge poses ample opportunities for biological upcycling, as the sludge contains high amounts of nutrients, energy and valuable biochemicals, making it an ideal food for extractive species. Sludge has been proposed as a feed stock for polychaete production, which in turn can be utilized live in shrimp aquaculture or as an aquafeed ingredient. However, the biosafety of such value chains has not yet been addressed. We conducted an experiment exposing the polychaete Hediste diversicolor to aquaculture sludge spiked with four different fish pathogens (Mycobacterium salmoniphilum, Yersinia ruckeri, Infectious Pancreatic Necrosis (IPN) and Infectious Salmon Anaemia (ISA)) known to cause diseases in Atlantic salmon (Salmo salar L.). Moreover, we assessed whether heavy metals and other potentially hazardous elements present in fish sludge bioaccumulates in the polychaetes. Neither of the bacteria nor viruses could be detected in the polychaetes after 14 days of continuous exposure. Seven of the 15 elements we analysed showed bioaccumulation factors significantly below one, meaning biodilution, while the other eight did not differ from one, meaning no bioaccumulation. None of the elements showed a significant bioaccumulation. Further on, none of the heavy metals found in the polychaetes at the end of our experiment exceeded the EU regulatory maximum levels for fish feed ingredients. The current results suggest that a H. diversicolor can reared on aquaculture sludge, and aquaculture sludge may serve as feed stock for polychaete production without the product exceeding EU regulations for contaminants in animal feed.

The green seaweed Ulva: tomorrow's "wheat of the sea" in foods, feeds, nutrition, and biomaterials.

Ulva, a genus of green macroalgae commonly known as sea lettuce, has long been recognized for its nutritional benefits for food and feed. As the demand for sustainable food and feed sources continues to grow, so does the interest in alternative, plant-based protein sources. With its abundance along coastal waters and high protein content, Ulva spp. have emerged as promising candidates. While the use of Ulva in food and feed has its challenges, the utilization of Ulva in other industries, including in biomaterials, biostimulants, and biorefineries, has been growing. This review aims to provide a comprehensive overview of the current status, challenges and opportunities associated with using Ulva in food, feed, and beyond. Drawing on the expertise of leading researchers and industry professionals, it explores the latest knowledge on Ulva's nutritional value, processing methods, and potential benefits for human nutrition, aquaculture feeds, terrestrial feeds, biomaterials, biostimulants and biorefineries. In addition, it examines the economic feasibility of incorporating Ulva into aquafeed. Through its comprehensive and insightful analysis, including a critical review of the challenges and future research needs, this review will be a valuable resource for anyone interested in sustainable aquaculture and Ulva's role in food, feed, biomaterials, biostimulants and beyond.

Influence of dietary bitter orange peel powder on growth, body composition, blood parameters, gut morphometry, and thermal tolerance of Nile tilapia (Oreochromis niloticus).

The potential of bitter orange peel powder (BOPP) as a nutritional strategy for fish was investigated in Nile tilapia. A total of 120 juveniles with an average initial weight of 9.8 ± 0.7 g were divided into four groups, replicated three times, resulting in 12 experimental units (60 L each) at a stocking density of 1.63 g of fish per liter. Productive parameters, whole-body composition, blood biochemistry, erythroid morphometry, intestinal histology, and heat tolerance were assessed in the juveniles subjected to one of the following treatments: non-supplemented basal diet (control group); basal diet with BOPP at 10 g/kg (BOPP10 group); basal diet with BOPP at 20 g/kg (BOPP20 group); and basal diet with BOPP at 40 g/kg (BOPP40 group). The BOPP additive had a positive influence on Nile tilapia growth, as final weight and weight gain were greater in all BOPP-treated fish, despite the reduction in crude protein in BOPP10 and BOPP20 groups. Fish receiving BOPP40 had an increase in total lipids and showed the highest levels of triglycerides and total cholesterol. Villi development was greater in the tilapia given BOPP10. It may be concluded that BOPP presented the most promising results for Nile tilapia juveniles when used at 10 g/kg diet. Regarding the erythroid morphometry, there was a general increase in nuclear and cytoplasmic areas in BOPP-fed tilapia; this seems to be the first report on the direct impact of the inclusion of functional additives in fish diet upon such parameters. As concerns the thermal tolerance evaluated at the end of the feeding trial, no differences were registered among the experimental groups. Thus, BOPP represents a feasible alternative ingredient to be explored in fish nutrition, since orange peel is a natural low-cost source of essential nutrients and valuable bioactive compounds.

Dietary supplementation of mineral nanoparticles for channel catfish (Ictalurus punctatus).

This study evaluated the supplementation of iron and copper nanoparticles in channel catfish diets and their influences on growth and health. A comparative feeding trial was carried out for 9 weeks to evaluate combinations of iron and copper nanoparticles: only iron nanoparticles (IronNP), only copper nanoparticles (CopperNP), CopperNP + IronNP, and a control diet supplemented with inorganic iron and copper (FeSO4 and CuSO4). After a 9-week feeding trial, growth performance, hematological parameters, whole-body proximate composition, and intestinal microbiota were evaluated, and fish were subjected to a bacterial challenge against Edwardsiella ictaluri to evaluate the contribution of the experimental treatments to fish health status. No statistical differences were detected for catfish fed the various diets in terms of production performance or survival after bacterial challenge. The hematocrit and RBC counts from fish fed the diet containing copper nanoparticles were significantly lower than the control group. A higher relative abundance of gram-positive bacteria was found in the digesta of catfish fed diets containing copper nanoparticles. Furthermore, in the context of hematology, iron nanoparticles did not impact the blood parameters of channel catfish; however, reduced hematocrits were observed in fish fed the copper nanoparticle diet, which lacked supplemental dietary iron, thus reinforcing the importance of dietary iron to catfish hematopoiesis. Nonetheless, additional studies are needed to investigate the effects of dietary copper nanoparticle supplementation in catfish diets to better illuminate its effects on the intestinal microbiota.

Economically viable bioprocess for inclusion of microalga Nannochloropsis gaditana in aquaculture feeds: Evaluation of antioxidant addition in preventing lipid oxidation during storage.

The rising prices of fishery derivatives limits their use in aquafeeds. Therefore, other alternatives are used to replace those ingredients. Among them, microalgae are of great interest both as an ingredient and as a potential stabilising agent against lipid oxidation. This study evaluates on the use of Nannochloropsis gaditana to prevent lipid oxidation in a set of 12 aquafeeds over 540 days of storage. Aquafeeds were formulated with/without 15 % N. gaditana combined with two antioxidants -butylhydroxytoluene (25-150 mg·kg-1) or vitamin E (500-3000 mg·kg-1). The effect of i) storage period, ii) presence of microalgae and iii) antioxidant addition on lipid oxidation was assessed. Results showed higher fatty acid degradation in diets lacking microalgae. The microalgae supplemented diets is enough for preserving feeds presenting the highest antioxidant effect at the end, without significant differences with the microalgae-supplemented feeds and those including antioxidants after 540 days of storage.

Insect meal in aquafeeds: A sustainable path to enhanced mucosal immunity in fish.

The mucosal surfaces of fish, including their intestines, gills, and skin, are constantly exposed to various environmental threats, such as water quality fluctuations, pollutants, and pathogens. However, various cells and microbiota closely associated with these surfaces work in tandem to create a functional protective barrier against these conditions. Recent research has shown that incorporating specific feed ingredients into fish diets can significantly boost their mucosal and general immune response. Among the various ingredients being investigated, insect meal has emerged as one of the most promising options, owing to its high protein content and immunomodulatory properties. By positively influencing the structure and function of mucosal surfaces, insect meal (IM) has the potential to enhance the overall immune status of fish. This review provides a comprehensive overview of the potential benefits of incorporating IM into aquafeed as a feed ingredient for augmenting the mucosal immune response of fish.

A Sustainable Diet for Tambaqui Farming in the Amazon: Growth Performance, Hematological Parameters, Whole-Body Composition and Fillet Color.

The aim of this study was to produce feed based on locally sourced ingredients for tambaqui farming in Amazon. Diets were formulated with increasing levels (0, 25, 50, 75 and 100%) of defatted black soldier fly larvae meal (BSFL) as a replacement for fish meal (FM), and cassava by-products in the same proportion (tuber residues, peel and leaves). A conventional diet (CO) was used as the control. Juvenile tambaqui (24.61 ± 1.14 g) were housed in 24 tanks in a recirculation aquaculture system. Neither diet rejection nor mortality were observed. Fish fed cassava by-products showed similar feed conversion rates (FCR 1.76); however, these values were worse than those observed in fish fed the CO (FCR 1.33). No differences were observed in the whole-body composition of the fish. The fillets of fish fed cassava by-products had a yellow color due the carotenoids present in the leaves. Dietary BSFL and cassava by-products can contribute to the sustainability of Amazonian aquaculture. Further studies with a lower proportion of cassava leaves in the diet formulation are recommended so as to ensure enhanced diet digestibility and less impact on the color of the fillets.

Use of grape by-products in aquaculture: New frontiers for a circular economy application.

Grape by-products have already been used in cosmetics, food industries, but also animal feed industry, especially monogastrics and in aquaculture. Grape by-products have been studied for a long time and their principal activities are antimicrobial and antioxidant. Concerning aquaculture, the great demand and necessity to replace animal sources with vegetable ones, has placed grape by-products as possible new phytonutrients with beneficial properties. The purpose of this review is to describe the use of grape by-products in aquaculture, during the last decade, concerning their effects on: 1) gut health and welfare status; 2) growth performances; 3) quality of fillets and flesh during the rearing cycle and shelf-life products. Although other studies highlighted that the high supplementation of grape by-products could negatively affect fish health and growth, due to antinutritional factors (tannins), grape by-products are proven to be valuable phytonutrients that can be incorporated into fish feed to enhance growth and health during rearing conditions. Even in fish products, their utilization has proven to elongate the properties and shelf-life of fillets and minces. Further studies to evaluate the possible integrations or replacements with grape by-products in fish feed in order to evaluate their effectiveness in aquaculture from a sustainable circular economy perspective will be desirable to enhance the use of these products.

Microalgae as fishmeal alternatives in aquaculture: current status, existing problems, and possible solutions.

Fishmeal is an indispensable ingredient for most aquatic animals. However, the finite supply and escalating price of fishmeal seriously limit its use in aquaculture. Thus the development of new, sustainable protein ingredients has been a research focus. Microalgae are potential fishmeal alternatives owing to their high protein content and balanced amino acid profile. Studies suggest that suitable replacement of fishmeal with microalgae is beneficial for fish growth performance, but excessive replacement would induce poor growth and feed utilization. Therefore, this paper aims to review research on the maximum substitutional level of fishmeal by microalgae and propose the main issues and possible solutions for fishmeal replacement by microalgae. The maximum replacement level is affected by microalgal species, fish feeding habits, quality of fishmeal and microalgal meals, and supplemental levels of fishmeal in the control group. Microalgae could generally replace 100%, 95%, 95%, 64.1%, 25.6%, and 18.6% fishmeal protein in diets of carp, shrimp, catfish, tilapia, marine fish, and salmon and trout, respectively. The main issues with fishmeal replacement using microalgae include low production and high production cost, poor digestibility, and anti-nutritional factors. Possible solutions to these problems are recommended in this paper. Overall, microalgae are promising fishmeal alternatives in aquaculture.

The application of protease in aquaculture: Prospects for enhancing the aquafeed industry.

Low-fishmeal and protein-saving diets are two prominent nutritional strategies utilized to address challenges related to the scarcity and sustainability of protein sources in aquaculture. However, these diets have been associated with adverse effects on the growth performance, feed utilization, and disease resistance of aquatic animals. To mitigate these challenges, exogenous protease has been applied to enhance the quality of diets with lower protein contents or fishmeal alternatives, thereby improving the bioavailability of nutritional ingredients. Additionally, protease preparations were also used to enzymatically hydrolyze fishmeal alternatives, thus enhancing their nutritional utilization. The present review aims to consolidate recent research progress on the use of protease in aquaculture and conclude the benefits and limitations of its application, thereby providing a comprehensive understanding of the subject and identifying opportunities for future research.

Long-term regulation of fat sensing in rainbow trout (Oncorhynchus mykiss) fed a vegetable diet from the first feeding: focus on free fatty acid receptors and their signalling.

Taste plays a fundamental role in an animal's ability to detect nutrients and transmits key dietary information to the brain, which is crucial for its growth and survival. Providing alternative terrestrial ingredients early in feeding influences the growth of rainbow trout (RT, Oncorhynchus mykiss). Thus, the present study aimed to assess the influence, via long-term feeding (from the first feeding to 8 months), of alternative plant ingredients (V diet for vegetable diet v. C diet for a control diet) in RT on the mechanism of fat sensing at the gustatory level. After the feeding trial, we studied the pathways of the fat-sensing mechanism in tongue tissue and the integrated response in the brain. To this end, we analysed the expression pattern of free fatty acid receptors (ffar) 1 and 2, markers of calcium-signalling pathways (phospholipase Cß, Orai, Stim or Serca), the serotonin level (a key neurotransmitter in taste buds) and the expression pattern of appetite-regulating neuropeptides in the hypothalamus (central area of appetite regulation). The results revealed that the V diet modified the expression pattern of ffar1 and paralogs of ffar2 genes in tongue tissue, along with differential regulation of calcium-signalling pathways and a defect in serotonin level and brain turnover, without influencing neuropeptide expression. This study is the first to support that changes in feeding behaviour of RT fed a V diet could be due to the difference in nutrient sensing and a decrease in hedonic sensation. We revealed that RT have similar fat-detection mechanisms as mammals.

Effect of digestible protein on intermediate metabolism, hepatic enzyme activities, energy reserves, and growth performance of pacu (Piaractus mesopotamicus) in the finishing growth phase.

This study investigated the effect of different levels of digestible protein (DP) on blood metabolites, hepatic enzyme activity of glycolysis and amino acid metabolism, energy reserves, and the production characteristics of pacu (Piaractus mesopotamicus) during the finishing growth phase. Six semi purified and isoenergetic diets, containing 16.3, 20.1, 23.8, 27.2, 31.5, and 34.8% of balanced DP, provided with essential amino acid balance, were hand-fed to pacu (1100.0 ± 10.3 g, initial weight) three times daily for 7 weeks. The experiment consisted of six treatments, with three randomly arranged replicates (tanks) per treatment. The data obtained from this experiment were analyzed by one-way analysis of variance (ANOVA), and significant differences (p < 0.05) between treatments were determined using Tukey's test. Blood metabolites, except serum ammonia and the hepatic enzymes activities of glycolysis and amino acid metabolism, except hexokinase activity were affected (p < 0.05) by balanced DP. The energy reserve indices, except hepatic total lipid content, were also found associated (p < 0.05) with balanced DP. The test diets significantly (p < 0.05) affected growth performance parameters. Higher dietary proteins led to a greater energy uptake by fish from the protein in feed. Overall, fish fed the intermediate level (23.8%) of balanced DP with digestible energy of 17.95 MJ kg-1 showed better production traits and physio-biochemical health markers. This information could help nutritionists and farmers to develop nutritionally balanced and economically and environmentally sustainable aquafeed for promoting healthy and sustainable production of pacu in intensive culture systems.

Insights into the functional properties of a natural free amino acid mix: Effect on growth performance, nutrient metabolism, and immune response in a carnivorous fish, Asian seabass (Lates calcarifer).

Dietary supplements containing a functional feed additive have been shown to be beneficial to fish and shellfish aquaculture. However, the functional properties of aquafeed formulations have rarely been reported in fish. This study aimed to investigate the effects of natural free amino acid mix (FAAM) supplementation as a functional solution on the growth performance and nutrient utilization in a carnivorous fish, Asian seabass (Lates calcarifer). Five isonitrogenous and isolipidic diets were prepared with graded supplementation levels of FAAM at 0 % (control group), 0.25 %, 0.50 %, 0.75 %, and 1.0 %, denoted as FAAM0, FAAM0.25, FAAM0.5, FAAM0.75, and FAAM1.0, respectively. The experimental fish were fed different dietary FAAM supplementations to apparent satiation twice daily for eight weeks. Significant improvements were observed in the growth performance of fish among the five groups (P < 0.05). Fish fed with FAAM0.75 displayed significantly increased activities of lysozyme, myeloperoxidase, catalase, and glutathione peroxidase (P < 0.05). The activities of digestive enzymes, including amylase, protease, and lipase, were enhanced by the supplementation of FAAM in the feed (P < 0.05), especially for the groups that contained more than 0.5 % FAAM in the feed. Furthermore, the morphological profile of the intestinal tract, including the mucosal fold height, width, thickness, and goblet cell, increased in fish fed with FAAM at 1.0 % (P < 0.05). Moreover, FAAM supplementation in diets not only modulated the expression of immune-related genes (glutathione peroxidase (GPx), complement (C)3, C4, and C-reactive protein) in the liver but also positively impacted the growth-ralated genes, including growth hormone (GH), GH receptor (GHR), insulin-like growth factor I (IGF-I), and IGF-II. In addition, the amounts of monounsaturated fatty acids (mainly oleic acid (C18:1n9c)) and polyunsaturated fatty acids-especially γ-linolenic acid (C18:3 n6) and α-linolenic acid (C18:3n3)-increased in fish fed with diets containing FAAMs (P < 0.05). Interestingly, the diets supplemented with FAAMs also had a positive effect on the economic indices in terms of revenue-to-cost ratios. These findings provide a scientific basis for the application of FAAMs as a functional solution that can be used in feed formulations for Asian seabass.

Combined Effects of Yellow Mealworm (Tenebrio molitor) and Saccharomyces cerevisiae on the Growth Performance, Feed Utilization Intestinal Health, and Blood Biomarkers of Nile Tilapia (Oreochromis niloticus) Fed Fish Meal-Free Diets.

Aquafeed quality is the most critical factor for aquaculture sustainability. However, limitations of traditional feed ingredients such as fishmeal (FM) need alternative strategies to ensure the nutritional requirements for aquatic animals. In this trial, four test diets were formulated (2 × 2 factorial design), where FM was incorporated in two diets at 10% with or without Saccharomyces cerevisiae (SC) at 1 g/kg. At the same time, FM was replaced with yellow mealworm (Tenebrio molitor) meal (TM) with or without SC at 1 g/kg. The growth performance indices (final weight, weight gain, and SGR), and the feed utilization indices (FCR and PER) were markedly affected by the protein source (FM or TM) and dietary SC (P < 0.05). The protein source (FM or TM) significantly (P < 0.05) affected the whole-body protein and lipid contents, while the moisture and ash contents were unaffected (P > 0.05) by TM or SC. The growth of the intestinal villi showed a marked increase in both height and branching in the treated groups with SC along the whole length of the intestine. Furthermore, the immune cell infiltration was prominent near the intestinal crypts of the middle intestinal segments in the supplemented groups by SC. Dietary TM and SC revealed improved hepatic parenchyma in the liver tissue better than other groups. The hematological indices, including hemoglobulin, hematocrit, red blood cells, and white blood cells, were markedly affected by dietary SC (P < 0.05). The lysozyme activity and phagocytic index were markedly affected by dietary SC, while phagocytic activity was affected by dietary TM (P < 0.05). The catalase, glutathione peroxidase, and malondialdehyde were markedly affected by the interaction between dietary protein source and SC, while superoxide dismutase was affected by dietary SC (P < 0.05). In conclusion, adding SC could enhance the utilization of TM by Nile tilapia with positive effects on the intestinal and liver histological features and the immune and antioxidative responses.

Effect of dietary gamma aminobutyric acid (GABA) modulated the growth performance, immune and antioxidant capacity, digestive enzymes, intestinal histology and gene expression of Nile tilapia (Oreochromisniloticus).

Dietary GABA plays an important role in a variety of physiological functions in animals, but this has been rarely reported in fish. This study investigated the effects of dietary supplementation with GABA on growth, serum antioxidant indicators, digestive enzyme activities, intestinal morphology, and the gene expression of Nile tilapia. Diets containing three different GABA concentrations, 0 (control), 200 mg/kg (GABA200), and 500 mg/kg (GABA500), were fed to Nile tilapia (6.97 ± 0.34 g) for 56 days. The present study found that increasing dietary GABA content could increase the fish growth performance including final body weight, weight gain, specific growth rate, average daily gain, protein efficiency ratio, and feed efficiency compared to the control diet. Interestingly, the feed conversion ratio was improved by dietary GABA supplementation. The antioxidant enzyme activities against ammonia stress of fish fed the GABA diets were significantly higher than the corresponding control group throughout the 96-h ammonia exposure. Moreover, significant increases in digestive enzyme activities including protease, amylase and lipase were found in fish fed the GABA diets. Intestinal morphology analysis revealed increased heights and widths of intestinal villi as well as thickness of the intestinal muscularis in fish fed the GABA diets compared to the control diet. The supplementation of diets with GABA significantly increased the expression level of immune- and growth-related genes. The above results indicate that dietary GABA can modulate the growth, improve their immune response and antioxidant status, gut health and morphology and gene expression of Nile tilapia. Therefore, GABA is a promising feed additive for Nile tilapia aquaculture.

Genetic Identification and Traceability of Insect Meals.

Insects have been proposed as a rich alternative source of protein for the partial or total replacement of fishmeal in aquaculture. For maximum safety and effectiveness of insect meals, control of the quality composition of these products is considered mandatory. The aim of this study was the genetic analysis of the composition of commercially available insect meals at the species level. Commercially available Hermetia illucens, Tenebrio molitor and Musca domestica individuals, as well as nine insect meals produced from these species, were analyzed. The genetic identification of insects at the species level was based on a COI fragment, and analysis of the insect meals' composition was performed with the processes of cloning and colony PCR. Genetic analysis indicated that the commercially available larvae morphologically identified as Musca domestica belonged to the species Muscina stabulans. In the commercially available insect meals, no other animal species was identified beyond the expected one. However, in the insect meal produced for research purposes, fungal growth was detected. The used methodology, herein, allows for the qualitative genetic identification of insect meals and could be included in the methods of traceability of products containing insects and other animal species.

Dietary Exogenous α-Amylase Modulates the Nutrient Digestibility, Digestive Enzyme Activity, Growth-Related Gene Expression, and Diet Degradation Rate of Olive Flounder (Paralichthys olivaceus).

In this study, a 12-week feeding experiment was conducted to characterize the effects of exogenous α-amylase on the growth, feed utilization, digestibility, plasma α-amylase activity, feed degradation rate, and fecal particle size of olive flounder (Paralichthys olivaceus). Diet was supplemented with 0 (AA0; control), 100 (AA100), 200 (AA200), or 400 (AA400) mg/kg of α-amylase, respectively. Fish (273.1 ± 2.3 g) were stocked into 12 tanks (25 fish/1,000-L tank) and 3 tanks were randomly selected for each diet group. As a result, α-amylase was found to have no significant effects (p ≥ 0.05) on the growth, feed utilization parameters, and whole-body proximate compositions. α-Amylase-treated fish exhibited only a significant increase in the apparent digestibility coefficient of carbohydrates compared to the controls. In addition, in vitro analyses revealed that α-amylase dose-dependently increased (p < 0.05) the feed degradation rate, while photographs of the intestinal content after 2, 4, and 8 h of feeding demonstrated an improved degradation rate in the α-amylase-treated groups. Plasma α-amylase content was higher in the AA200 and AA400 groups, whereas the control group produced significantly larger-sized fecal particles (90% size class) than these two groups. In the intestine, no changes were observed in the expression levels of the immune-related TNF-α, IL-1ß, IL-2, immunoglobulin-M, HSP-70, lysozyme, and amylase alpha-2A. However, growth-related genes IGF-1, IGF-2, TGF-ß3, and growth hormone genes were upregulated in muscle tissues. Collectively, exogenous α-amylase has positive roles in the modulation of the digestibility coefficient, blood α-amylase concentration, growth-related gene expression, and diet degradation for improved digestion in olive flounder.

Evaluation of the Combined Administration of Chlorella fusca and Vibrio proteolyticus in Diets for Chelon labrosus: Effects on Growth, Metabolism, and Digestive Functionality.

This study aimed to evaluate the combined effect of dietary Chlorella fusca and ethanol-inactivated Vibrio proteolyticus DCF12.2 (C + V diet) in Chelon labrosus juveniles, highlighting their nutritional, physiological, and morphological effects. The results showed that the combined dietary inclusion of C. fusca and V. proteolyticus significantly enhanced growth performance and feed utilization compared to the control group. The C + V diet increased the fish lipid quality index (FLQ), n-3 polyunsaturated fatty acids, and n-3/n-6 ratio, which might be beneficial in terms of human nutrition. The C + V diet considerably increased carbohydrate metabolic activity by statistically boosting plasma glucose. The dietary inclusion of C. fusca in conjunction with V. proteolyticus increased metabolic enzyme activity as well as intestinal absorption capacity compared to that found in the control group. In conclusion, the experimental diet was suitable for feeding C. labrosus, increasing their growth and the nutritional characteristics of the muscle and intestine, without causing tissue damage.