Search

The aim of the present work was to investigate the influence of fasting and refeeding on body condition, gut physiology and microbiota in reared O. mykiss. Ninety-six fish were randomly allotted among three groups subjected to different feeding plan: C (control, fed for 5 weeks); R (restricted ration over 3 weeks followed by 2 weeks feeding); F (fasted over 3 weeks followed by 2 weeks feeding) in a well’s fresh water flow-through rearing plan. Sampling occurred at 0, 1, 2, 4, 7, 14 days during the refeeding period. At day 0 and throughout the feeding period until day 14, the weight of the fish was significantly affected by the feeding restriction. Feed deprivation reduced significantly the viscerosomatic and hepatosomatic indexes. Brush border membrane enzymes’ specific activity was modulated by feeding regimes until day 7, to level in all experimental groups at day 14. At the end of the restricted/fasted period, the microbiota of the C group was made up of 70% of Actinobacteria, 24% of Proteobacteria, 4.2% of Firmicutes and

One of the main sources of MPs contamination in fish farms is aquafeed. The present study investigated, for the first time through a comparative approach, the effects of different-sized fluorescent MPs included in a diet intended for zebrafish (Danio rerio). A comparison based on fish developmental stage (larval vs. juvenile), exposure time, and dietary MPs’ size and concentration was performed. Four experimental diets were formulated, starting from the control, by adding fluorescent polymer A (size range 1–5 µm) and B (size range 40–47 µm) at two different concentrations (50 and 500 mg/kg). Zebrafish were sampled at 20 (larval phase) and 60 dpf (juvenile stage). Whole larvae, intestine, liver and muscles of juveniles were collected for the analyses. Polymer A was absorbed at the intestinal level in both larvae and juveniles, while it was evidenced at the hepatic and muscular levels only in juveniles. Hepatic accumulation caused an increase in oxidative stress markers in juveniles, but at the same time significantly reduced the number of MPs able to reach the muscle, representing an efficient barrier against the spread of MPs. Polymer B simply transited through the gut, causing an abrasive effect and an increase in goblet cell abundance in both stages.

Within the modern aquaculture goals, the present study aimed to couple sustainable aquafeed formulation and culturing systems. Two experimental diets characterized by 3 and 20% of fish meal replacement with full-fat spirulina-enriched black soldier fly (Hermetia illucens) prepupae meal (HPM3 and HPM20, respectively) were tested on European seabass (Dicentrarchus labrax) juveniles during a 90-day feeding trial performed in aquaponic systems. The experimental diets ensured 100% survival and proper zootechnical performance. No behavioral alterations were evidenced in fish. Histological and molecular analyses did not reveal structural alterations and signs of inflammation at the intestinal level, highlighting the beneficial role on gut health of bioactive molecules typical of HPM or derived from the enriching procedure of insects’ growth substrate with spirulina. Considering the quality traits, the tested experimental diets did not negatively alter the fillet’s fatty acid profile and did not compromise the fillet’s physical features. In addition, the results highlighted a possible role of spirulina-enriched HPM in preserving the fillet from lipid oxidation. Taken together, these results corroborate the use of sustainable ingredients (spirulina-enriched HPM) in aquaponic systems for euryhaline fish rearing.

In the present study, an organic substrate (coffee silverskin) enriched with spirulina (

The production of insects on an industrial scale has attracted the attention of the research and agricultural industry as novel protein sources. To detect the presence of Gryllodes sigillatus (GS) in feed and food, a real-time PCR method based on the mitochondrial cytochrome b (CYB) gene is proposed by this study. Forty DNA samples of animal and plant origin were used to confirm the specificity of the qPCR system. The detection method’s performance was evaluated on different processed GS matrices including native GS (UnGS) and different commercial products: crunchy roasted samples (RoGS), insect meal mixtures (ACGS) and energetic snacks containing GS (GSS). Data on sequencing were aligned with the reference gene to confirm the PCR products. The regression curve (y = −3.394 x + 42.521; R2 = 0.994, d.f. 14) between Ct values and Log DNA concentrations of Gryllodes sigillatus resulted in an efficiency of 96.4%. The severity of the technological processing treatments and the matrix structure affected the intensity of the PCR signal with the same amount of insect DNA as observed by different y-intercepts of the three-regression lines for RoGS, ACGS, and GSS. The real-time PCR method resulted in robust and sensitive outcomes able to detect low amounts of GS DNA (5 g/100 g) in a complex matrix, making it suitable for detecting the presence or absence of labeled Gryllodes sigillatus material both in feed and food.

Insects represent a valuable and sustainable alternative ingredient for aquafeed formulation. However, insect-based diets have often highlighted controversial results in different fish species, especially when high inclusion levels were used. Several studies have demonstrated that nutritional programming through parental feeding may allow the production of fish better adapted to use sub-optimal aquafeed ingredients. To date, this approach has never been explored on insect-based diets. In the present study, five experimental diets characterized by increasing fish meal substitution levels with full-fat Black Soldier Fly (Hermetia illucens, BSF) prepupae meal (0%, 25%, 50%, 75% and 100%) were used to investigate the effects of programming via broodstock nutrition on F1 zebrafish larvae development. The responses of offspring were assayed through biometric, gas chromatographic, histological, and molecular analyses. The results evidenced that the same BSF-based diets provided to adults were able to affect F1 zebrafish larvae fatty acid composition without impairing growth performances, hepatic lipid accumulation and gut health. Groups challenged with higher BSF inclusion with respect to fish meal (50%, 75% and 100%) showed a significant downregulation of stress response markers and a positive modulation of inflammatory cytokines gene expression. The present study evidences that nutritional programming through parental feeding may make it possible to extend the fish meal substitution level with BSF prepupae meal in the diet up to almost 100% without incurring the well-known negative side effects of BSF-based diets.

Simple Summary Sustainability and fish welfare have been receiving increasing attention in the aquaculture sector, with an emphasis on the search for new, sustainable, and healthy aquafeed ingredients. For many years, plant ingredients have been widely used in aquafeed formulation; however, negative side effects on gut welfare have often been reported in several carnivorous fish species. From this perspective, alternative ingredients such as poultry by-products and insect meal are receiving attention due to their low ecological footprint and high nutritional value. In the present study, these two ingredients were used, singly or in combination, to formulate practical diets for gilthead seabream (Sparus aurata). After a twelve-week feeding trial, a multidisciplinary laboratory approach including histological, molecular, and spectroscopic techniques was adopted in order to investigate fish physiological responses to the new test diets. The results obtained showed excellent zootechnical performances and ameliorated gut health in fish fed dietary inclusions of poultry by-products and insect meal compared to those fed a vegetable-based diet. In addition, the modulation of nutrient absorption in relation to the ingredients used was highlighted by means of spectroscopic tools. The results obtained demonstrated that poultry by-products and insect meal can be successfully used to replace plant-derived ingredients in diets for gilthead seabream without negatively affecting fish welfare. Abstract The attempt to replace marine-derived ingredients for aquafeed formulation with plant-derived ones has met some limitations due to their negative side effects on many fish species. In this context, finding new, sustainable ingredients able to promote fish welfare is currently under exploration. In the present study, poultry by-products and Hermetia illucens meal were used to replace the vegetable protein fraction in diets totally deprived of fish meal intended for gilthead seabream (Sparus aurata). After a 12-week feeding trial, a multidisciplinary approach including histological, molecular, and spectroscopic techniques was adopted to investigate intestine and liver responses to the different dietary formulations. Regardless of the alternative ingredient used, the reduction in dietary vegetable proteins resulted in a lower incidence of intestine histological alterations and inflammatory responses. In addition, the dietary inclusion of insect meal positively affected the reduction in the molecular inflammatory markers analyzed. Spectroscopic analyses showed that poultry by-product meal improved lipid absorption in the intestine, while insect meal induced increased liver lipid deposition in fish. The results obtained demonstrated that both poultry by-products and H. illucens meal can successfully be used to replace plant-derived ingredients in diets for gilthead seabream, promoting healthy aquaculture.

By answering the need for increasing sustainability in aquaculture, the present study aimed to compare growth, gene expression involved in appetite regulation, physical characteristics, and chemical composition of Sparus aurata fed alternative protein sources. Fish were fed ten iso-proteic, iso-lipidic, and isoenergetic diets: a vegetable-based (CV) and a marine ingredient-rich (CF) diet were set as control diets. The others were prepared by replacing graded levels (10, 20 or 40%) of the vegetable proteins in the CV with proteins from a commercial defatted Hermetia illucens pupae meal (H), poultry by-product meal (PBM) singly (H10, H20, H40, P20, P40) or in combination (H10P30), red swamp crayfish meal (RC10) and from a blend (2:1, w:w) of Tisochrysis lutea and Tetraselmis suecica (MA10) dried biomasses. The increase in ghre gene expression observed in MA10 fed fish matched with increased feed intake and increased feed conversion ratio. Besides, the MA10 diet conferred a lighter aspect to the fish skin (p &lt, 0.05) than the others. Overall, no detrimental effects of H, PBM, and RC meal included in the diets were observed, and fish fatty acid profile resulted as comparable among these groups and CV, thus demonstrating the possibility to introduce H, PBM, and RC in partial replacement of vegetable proteins in the diet for Sparus aurata.

In the last decades, processed animal proteins, such as poultry by-product meal (PBM) and insect meals have received great attention as sustainable and nutritious aquafeed ingredients. The aim of the present study was to evaluate growth performances, liver and gut histology, macromolecular composition and inflammatory response in rainbow trout (Oncorhynchus mykiss) fed diets deprived of fish meal, where graded levels of vegetable protein-rich ingredients, were replaced by defatted Hermetia illucens pupae meal (HM) or PBM, singly or in combination. To this end eight grossly iso‐proteic (45% DM), isolipidic (26% DM) and isoenergetic (23.5 MJ/kg DM) were offered each to triplicated groups of juveniles' fish in a 91 days feeding trial. A diet rich in vegetable protein derivatives high in soybean meal (CV) was prepared to have a 10:90 and 20:80 fish to vegetable protein and lipid ratios respectively. By contrast, a fish-based diet (CF) was formulated with opposite fish to vegetable protein and lipid ratios. Six more diets, were obtained by replacing graded levels of protein (10, 30 and 60%) of diet CV, by protein from a defatted Hermetia illucens pupae meal and/or poultry by-product meal, singly or combined, while maintaining the same vegetable to fish lipid ratio as in the CV diet. Relative to diets CV and CF, a medium to high substitution (30 and 60%) of dietary vegetable protein-rich ingredients, with HM and/or PBM resulted in improved growth performance as well as in a minor incidence of distal intestine morphological alterations. The diet including both the test animal proteins led to nearly the best overall response in terms of growth and gut/liver health. Both HM and PBM when included at moderate or high levels in the diet, resulted in a downregulation of the expression of inflammatory-related genes relative to diet CV. This effect was greater with HM than that observed with PBM and goes beyond the parallel reduction of vegetable protein and SBM levels in the same diets, suggesting a beneficial role of insect meal that warrant further investigation. The results obtained so far, provide support to a reliable use of alternative/underexploited protein and lipid sources [(HM) or (PBM)] in developing a new generation of sustainable and healthy trout diets that meet the circular economy principles.

Over the last years, the potential use of Black Soldier Fly meal (BSF) as a new and sustainable aquafeed ingredient has been largely explored in several fish species. However, only fragmentary information is available about the use of BSF meal-based diets in sturgeon nutrition. In consideration of a circular economy concept and a more sustainable aquaculture development, the present research represents the first comprehensive multidisciplinary study on the physiological effects of a BSF diet during sturgeon culture in an aquaponic system. Siberian sturgeon (Acipenser baerii) juveniles were fed over a 60-days feeding trial on a control diet (Hi0) and a diet containing 50% of full-fat BSF meal respect to fish meal (Hi50). Physiological responses of fish were investigated using several analytical approaches, such as gas chromatography-mass spectrometry, histology, Fourier Transformed Infrared Spectroscopy (FTIR), microbiome sequencing and Real-time PCR. While aquaponic systems performed optimally during the trial, Hi50 group fish showed lower diet acceptance that resulted in growth and survival reduction, a decrease in hepatic lipids and glycogen content (FTIR), a higher hepatic hsp70.1 gene expression and a worsening in gut histological morphometric parameters. The low feed acceptance showed by Hi50 group sturgeon highlighted the necessity to improve the palatability of BSF-based diet designed for sturgeon culture.

Black Soldier Fly (BSF) meal is considered an alternative, emerging, and sustainable ingredient for aquafeed formulation. However, results on fish physiological responses are still fragmentary and often controversial, and no data are available on the effect of insect meal-based diets on fish reproduction. On this regard, zebrafish, with its relatively short life cycle, represents an ideal experimental model to explore this topic. In this study, female zebrafish were fed for 12 months on a control diet based on fish meal (FM) and fish oil and two experimental diets with full-fat BSF (

Black Soldier Fly (BSF) meal is considered as an alternative, emerging and sustainable ingredient for aquafeed production. However, results on fish physiological responses are still fragmentary and often controversial, while no studies are available on fish behavior in response to these new diets. The present work represents the first comprehensive multidisciplinary study aimed to investigate zebrafish physiological and behavioural responses to BSF-based diets. Five experimental diets characterized by increasing inclusion levels (0, 25, 50, 75 and 100% respect to fish meal) of full fat BSF prepupae meal were tested during a 2-months feeding trial. Prepupae were cultured on coffee silverskin growth substrate enriched with a 10% Schizochytrium sp. to improve insects’ fatty acids profile. The responses of zebrafish were assayed through biometric, histological, gas chromatographic, microbiological, spectroscopic, molecular and behavioural analyses. Results evidenced that BSF-based diets affected fish fatty acid composition, while behavioural tests did not show differences among groups. Specifically, a 50% BSF inclusion level diet represented the best compromise between ingredient sustainability and proper fish growth and welfare. Fish fed with higher BSF inclusions (75 and 100%) showed hepatic steatosis, microbiota modification, higher lipid content, fatty acid modification and higher expression of immune response markers.

Promoting circularity in the aquaculture sector through the conversion of great amount of organic by-products produced on land is a valuable strategy for a further development of the aquaculture sector. In this regard, insects represent a very promising example of bio-converting organisms; their application in aquafeeds, however, still faces possible limitations because of their lack in polyunsaturated fatty acids and the presence of chitin. The aim of the present study was to apply circularity to Black Soldier Fly (BSF) (Hermetia illucens) culture and to improve the insect's biomass fatty acid composition by culturing them on a land-produced by-product (coffee silverskin) enriched with a 10% Schizochytrium sp. The insect biomass was then used to formulate five fish diets containing 0, 25, 50, 75 and 100% of insect meal respect to fish meal, respectively. Diets were used for a feeding trial during zebrafish (Danio rerio) larval development (21 days) and a multidisciplinary approach including biometry, histology, gas chromatography, spectroscopy (FTIR), microbiota analyses and molecular biology was applied to better understand fish responses to the new diets. Results showed that the 50% substitution of fish meal with insect meal represented the best compromise between ingredient sustainability and proper fish growth and welfare. Fish fed with higher BSF inclusions (75 and 100%) showed a severe degree of hepatic steatosis, microbiota modification, a higher lipid content (FTIR), fatty acid modification and higher expression of both stress and immune response markers.

Insects are able to bio-convert organic by-products into a sustainable biomass for aquafeed formulation. Specifically, among several insect species, Hermetia illucens (H) is particularly interesting for its nutritious traits but, unfortunately, the lipidic fraction is poorly represented by polyunsaturated fatty acids n-3 and poses some limits in its application in aquafeed formulation. The present study undertook an interdisciplinary approach to explore the effects of three experimental diets containing increasing levels of full-fat H meal (H0 diet based on fishmeal and purified protein-rich vegetable ingredients; H25 and H50 diets containing 25% or 50% of full-fat H meal replacing fishmeal, respectively), on rainbow trout (Oncorhynchus mykiss) fed over a 98 days experimental period. The expression of genes related to lipid metabolism by RT-qPCR, liver histology, as well as the qualitative traits of fillets and fatty acid (FA) composition were investigated. Interestingly, fads2 gene expression in pyloric caeca increased in fish fed diets containing the highest full-fat H meal inclusion (H50 > H0; p The overall results showed that the dietary full-fat H meal inclusion under study did not impair fish fillet quality, guaranteeing its nutritional value. Some effects on lipid metabolism were observed, as suggested by liver, pyloric caeca and mid intestine gene expression and liver FA profile. Future studies on the biological mechanisms behind the macroscopic traits of fish fed unprocessed insects are warmly encouraged.

Black Soldier Fly (BSF) meal is considered an alternative, emerging, and sustainable ingredient for aquafeed formulation. However, results on fish physiological responses are still fragmentary and often controversial, and no data are available on the effect of insect meal-based diets on fish reproduction. On this regard, zebrafish, with its relatively short life cycle, represents an ideal experimental model to explore this topic. In this study, female zebrafish were fed for 12 months on a control diet based on fish meal (FM) and fish oil and two experimental diets with full-fat BSF (Hermetia illucens) prepupae meal inclusion, to replace 25% and 50% of FM (BSF25 and BSF50). All diets were isonitrogenous, isolipidic, and isoenergetic. The effects of these two experimental diets on female's reproduction were investigated through a multidisciplinary approach, including the evaluation of growth, gonadosomatic index, spawned/fertilized eggs and hatching rate, adult female carcass and fertilized egg fatty acid composition, histological analysis of the ovary, spectroscopic macromolecular composition of class IV oocytes, and expression of genes involved in fish lipid metabolism in the liver. Results showed that while fish were perfectly able to cope with a 25% insect meal dietary inclusion, a 50% inclusion level caused the overexpression of genes involved in lipid metabolism, a general reduction in the number of spawned eggs, and differences in the frequency rate of previtellogenic oocytes, class III, IV, oocytes and postovulatory follicles and atretic oocytes, in the macromolecular composition of class IV oocytes, and in the fatty acid composition of the fertilized eggs, respect to control and 25% group.

The aim of this study was to evaluate nutrient digestibility, growth response, carcass and fillet yields, muscle tissue composition and skin colour appearance of European sea bass (D. labrax) fed graded levels of a mixture of Tisochrysis lutea and Tetraselmis suecica freeze-dried biomass in partial substitution for fish meal and oil in diets containing substantial levels of vegetable oils and protein-rich derivatives. Five complete diets were formulated to be grossly isoproteic and isolipidic and prepared by including a blend of the two freeze-dried microalgae biomass in a 2:1 weight ratio, to replace approximately 15, 30 and 45% fish meal protein and 12, 24 and 36% fish lipid of a positive control diet. A negative control complete feed high in soybean meal was also prepared. Each diet was offered in triplicate, during 105 days until visual satiety of fish (204 ± 12.7 g) kept in a semi-closed recirculating marine water system ensuring optimal water quality to E. sea bass. The results of the study have shown that replacing about 45% crude protein and 36% lipid from fish meal and lipid by a mixture of Tisochrysis lutea and Tetraselmis suecica dried biomass, did not adversely affect growth performance and feed conversion efficiency of European sea bass. A slight decline in dry matter, protein and energy digestibility occurred in response to graded levels of dietary microalgae biomass, which was compensated by increased feed intake. Moreover, the diet including the dried microalgae resulted in a higher nutritive value than that of the negative high-soybean meal control feed. No major changes were observed in biometry traits and slaughter yield while the nutritional properties of the edible muscle tissue were little affected in terms of n-3 PUFA. The presence of the dried microalgae in the diet resulted in a greenish pigmentation of the skin, with a slight tendency towards redness and diminished lightness and hue. Overall the results obtained here reveal a potential improvement in sustainability in terms of reduced reliance on halieutic feed resources in case of the microalgae-containing diets. Hence the sustainable use of the dried mixture of the 2 marine microalgae biomass as feed ingredients in diets for the E. sea bass seems to be mainly limited by the low availability and unaffordable market price which are both expected to improve over the next 10 years.

Despite a growing interest in microalgae and cyanobacteria as potential sources of nutrients in aquafeeds, little information is presently available on their nutritive value for carnivorous fish species. The aim of this study was to evaluate chemical composition and nutrient digestibility of a panel of microalgae and cyanobacteria dried biomasses (MACB), using rainbow trout (Oncorhynchus mykiss W.) as a fish model. Nine test diets were obtained by mixing 80 parts of a reference diet, added with 20 g/kg of acid insoluble ash as an indigestible marker, to 20 parts of each of the following dried whole-cell biomass: Arthrospira platensis, Nostoc sphaeroides, two strains of Chlorella sorokiniana, Nannochloropsis oceanica, Tisochrysis lutea, Phaeodactylum tricornutum, Porphyridium purpureum and Tetraselmis suecica. The digestibility measurements were conducted with rainbow trout (52.4 ± 1.5 g) kept in six tank units each including three 60-L vessels singularly stocked with 12 fish and fitted with a settling column for faecal recovery. Per each diet, faeces were collected over three independent 10-day periods. Apparent digestibility coefficients (ADCs) of dry matter, crude protein (CP), organic matter and gross energy (GE) of single MACB were calculated by difference relative to those of the reference diet. The MACBs had heterogeneous chemical composition (CP, from 20 to 69%; Lipid, 5-27%; GE, 12.5--22.6 MJ/kg dry matter basis) reflecting their overall biodiversity. Most of them can be considered as virtually good sources of minerals and trace elements and exhibit an essential amino acid profile comparable or even better than that of soybean meal commonly used in fish feeds with P. purpureum showing the best protein profile. The digestibility results obtained with rainbow trout allowed ranking the MACBs into two major groups. A first one, including C. sorokiniana, N. oceanica and T. suecica, resulted in markedly lower (P < 0.05) crude protein and energy ADC (64-73%; 51-59%, respectively) compared to a second group including P. purpureum, T. lutea and cyanobacteria (CP-ADC, 83-88%; GE-ADC, 74-90%) while P. tricornutum resulted in intermediate values. Overall, the present study confirms the consistently reported role of cell-wall structure/composition in affecting accessibility of nutrients to digestive enzyme. Based on the overall outcomes, only T. lutea and cyanobacteria actually meet the requirements for being used as protein sources in aquafeeds provided their mass production becomes more feasible and cost-effective, hence attractive for the feed-mill industry in the near future.

Antibiotic resistance (AR) is an issue in aquaculture since the excessive use of antibiotics administered both for prophylaxis and metaphylaxis may lead to selection for resistant bacteria, that can be found in edible products (e.g., seafood) marketed for human consumption. There is a paucity of data in the scientific literature on the dynamics of AR genes in fish feeds, including those with insect meal as ingredient. This study represents the first attempt to assess the dynamics of genes conferring resistance to antibiotics conventionally used in clinical practice such as macrolide-lincosamide-streptogramin B (MLSB) [erm(A), erm(B), erm(C)], vancomycin (vanA, vanB), tetracyclines [tet(M), tet(O), tet(S), tet(K)], β-lactams (mecA, blaZ) and aminoglycosides [aac(6’)-Ie aph(2”)-Ia] (abbreviated as aac-aph). For this purpose, zebrafish (Danio rerio) fed Hermetia illucens-based diets was used as an experimental model. AR genes were assessed using optimized quantitative PCR (qPCR) assays. The dynamics of the selected AR genes were studied during a 6-month feeding trial with zebrafish fed diets including increasing levels of H. illucens (Hi) larvae (25 %, 50 %, 75 %, and 100 %) that were previously reared on coffee silverskin supplemented with 10 % Schizochytrium sp. microalgae. A diet with no insect additions was used as the control. qPCR detected erm(B), tet(K), tet(M), tet(O) and tet(S) genes in all the analyzed diet samples, whereas the same samples were negative for mecA, vanA, vanB and aac-aph. In zebrafish at their larval, juvenile and reproductive stages, erm(A), erm(C), vanB and aac-aph genes were never detected, whereas erm(B), tet(M) and tet(S) genes were widespread irrespective of the stage of development or feed. According to principal component analysis, a clear separation among fish fed diets containing the highest or the lowest insect inclusion levels was generally observed. These results showed a significant contribution of insect-based feed to the spread of the studied AR genes compared with control diets. This finding suggests the occurrence of complex interactions between the microbiota and the resistome occurring in feed and in the fish gut. Notably, when present, AR genes could be transferred to commensal or pathogenic microorganisms that may reach the food chain. These results represent a first baseline for future AR risk assessments in the edible insect feed chain. Until then, prudent use of antibiotics during rearing of edible insects is recommended.

Sustainability of aquaculture is tied to the origin of feed ingredients. In search of sustainable fish meal-free formulations for rainbow trout, we evaluated the effect of Hermetia illucens meal (H) and poultry by-product meal (P), singly (10, 30, and 60% of either H or P) or in combination (10% H + 50% P, H10P50), as partial replacement of vegetable protein (VM) on gut microbiota (GM), inflammatory, and immune biomarkers. Fish fed the mixture H10P50 had the best growth performance. H, P, and especially the combination H10P50 partially restored α-diversity that was negatively affected by VM. Diets did not differ in the Firmicutes:Proteobacteria ratio, although the relative abundance of Gammaproteobacteria was reduced in H and was higher in P and in the fishmeal control. H had higher relative abundance of chitin-degrading Actinomyces and Bacillus, Dorea, and Enterococcus. Actinomyces was also higher in H feed, suggesting feed-chain microbiome transmission. P increased the relative abundance of protein degraders Paeniclostridium and Bacteroidales. IL-1β, IL-10, TGF-β, COX-2, and TCR-β gene expression in the midgut and head kidney and plasma lipopolysaccharide (LPS) revealed that the diets did not compromise the gut barrier function or induce inflammation. H, P, and H10P50 therefore appear valid protein sources in fishmeal-free aquafeeds.

The aim of this study was to investigate the effects of replacing graded levels of dietary fish meal by a blend of two marine microalgae Tisochrysis lutea and Tetraselmis suecica on intestinal morpho-physiology and innate immune response in European sea bass. Two complete diets were formulated to be iso-nitrogenous and isolipidic and prepared by including a blend of the two microalgae, to replace approximately 15 and 45% fish meal protein of the control diet. A fourth diet, where the microalgae mix was substituted by soybean meal, was also prepared. Each diet was offered until visual satiety over 105 days to triplicated groups of European sea bass (204 ± 12.7 g), kept in a recirculating marine water system. The humoral and cellular innate immune parameters of E. sea bass were affected by the dietary treatment. Fish fed the microalgae-containing or the soybean rich diets, showed a significantly greater villi height, while the thickness of intestinal epithelium was significantly reduced in fish fed the soybean meal-rich diet. The activity of the brush border membrane enzymes, maltase, sucrase-isomaltase, γ-glutamil transferase and alkaline phosphatase was not affected by dietary treatment but changed in different intestinal tracts. The genes sucrase-isomaltase, peptide transporter 1, sodium/potassium-transporting ATPase and aminopeptidase N were overexpressed in the pyloric and proximal region of the intestine of fish fed the microalgae-including diets. In conclusion, a blend of dried marine microalgae Tisochrysis lutea and Tetraselmis suecica as alternative ingredients to dietary fish meal did not hamper gut digestive-absorptive functions of E. sea bass. Moreover, it resulted in enhanced non-specific immune response, suggesting an effective role as an immunostimulant ingredient.

A desirable goal of the aquaculture sector is to replace most of fish meal and fish oil with more sustainable, cost-effective, and environmental friendly ingredients ensuring fish health and welfare standards. Due to minimal environmental impact, compared with most conventional feed commodities, insects deserve a growing attention as candidate ingredients for aquafeeds. The present study investigated, for the first time, the possible application of a 100% insect diet in zebrafish larval rearing. Through a multidisciplinary approach, the major biological responses of fish to the new diets were assessed. Results of biometry, fatty acid composition, expression of genes involved in fish growth, stress response, lipid metabolism, chitinolytic activity, gut inflammation, and liver macromolecular composition suggested a possible application of insect larvae for zebrafish larval rearing. However, further studies are necessary to better understand the use of this insect species in the rearing of fish.

Fish intestine plays various physiological functions and its integrity is essential to guarantee fish growth and welfare. Several laboratory techniques have been proposed for studying morpho-physiology of fish intestine, but classical histology still represents one of the gold standards. Nevertheless, this technique is time consuming and does not provide comprehensive information on the biochemical composition of tissues. The present study, introduces a new approach for studying fish intestine, the Fourier Transform Infrared (FTIR) spectroscopy. This is a fast, label-free analytical technique, which analyses the vibrational transitions induced on tissues by the interaction with the electromagnetic radiation. On the base of vibrational features of some functional groups classes and molecular bonds, FTIR allows the analysis of the macromolecular composition of the sample. Rainbow trout (O. mykiss) represents an important species for modern aquaculture and deeper knowledge about its intestine anatomy and macromolecular composition is desirable. In the present study, besides the traditional histological analysis, the biochemical building of the intestine mucosa was analyzed for the first time by FTIR. Spectral data can be considered a complement to traditional histological analysis.

Due to minimal environmental impact, compared to most conventional feed commodities, insects deserve a growing attention as candidate ingredients for aquafeeds. This study tested, for the first time during zebrafish larval rearing, the effects of an increasing replacement (0%-25%-50%) of fish meal by black soldier fly (BSF) full-fat prepupae meal. All diets were formulated to be isonitrogenous and isolipidic. A multidisciplinary approach, including biometrics, histology, gas chromatography-mass spectrometry, and molecular analyses, was applied to better understand the biological responses of larval zebrafish to the different partial inclusions of BSF in the feed. Generally, results are promising, but a 50% of BSF meal inclusion in the diet affected both lipid composition and accumulation in the larvae.

This study investigated the effects of dietary inclusion levels of full-fat Hermetia illucens prepupae meal (H) on growth and gastrointestinal integrity in rainbow trout (Oncorhynchus mykiss). A 98-day study was conducted using triplicate groups of trout (initial body weight, 137 &plusmn, 10.5 g) kept in 1-m3 tanks in a flow-through well water system. Three dietary treatments were prepared: one based on fishmeal and purified protein-rich vegetable ingredients (H0), and two experimental diets including graded levels of H meal (25% and 50%, referred to as H25 and H50, respectively). At the end of the feeding trial, no differences were observed in growth performance and plasma metabolite levels, with the biometric data confirmed by the liver expression of the genes involved in somatic growth regulation (igf1 and mstn1a). In the H50 group, a three-fold up regulation of liver hsp70 was observed. An activation of the stress/immune response (il-10, tnf-&alpha, and tlr-5) was observed in medium intestine in the H25 and H50 groups (p &lt, 0.05) together with a villi length reduction detected through histological analyses. Liver histology and Fourier Transform Infrared Imaging (FTIRI) spectroscopy highlighted an increase in lipid deposition. These findings suggest that caution should be taken into account when 50% replacement of conventional ingredients with H is selected.

The objective of the present study was to evaluate the modulation of acute stress response by dietary nucleotides (NT) in sole, Solea solea. A basal diet was supplemented with levels of 0 (normal diet), or 0.4 g NT/kg dry diet for 8 weeks. At the end of feeding trial, fish fed the normal and NT-supplemented diet were subjected to a standardized protocol of disturbance and sampled over a 24h recovery after the stressor exposure. Modulatory effects of NT on acute stress response (cortisol and glucose), proopiomelanocortin (POMC) and cannabinoid receptor 1 splice variants (CB1A and CB1B) mRNA levels were studied. Both plasma cortisol and glucose levels of fish fed NT-supplemented diet were significantly lower than fish fed the control diet at 1 and 4h post-stress time-points. There are no significant effects of dietary NT on POMC and HSP70 mRNA levels. In our study, both CB1A and CB1B trascript levels were induced in fish fed the normal diet at 1 and 4h post-stress intervals. Collectively, the results obtained suggest that dietary NT modulates the CB1-like receptor mRNA expressions leading to attenuation in stressor-induced plasma cortisol level in sole.

Considering the well-known problems arising from the use of rotifers and Artemia as live prey in larval rearing in terms of fatty acid deficiencies, the aim of this study was to evaluate a partial or complete replacement of traditional live prey with preserved copepods during the larviculture of gilthead sea bream (Sparus aurata). Sea bream larvae were randomly divided into 4 experimental groups in triplicates: group A larvae (control) fed rotifers followed by Artemia nauplii; group B fed a combined diet (50%) of rotifers–Artemia and preserved copepods; group C fed rotifers followed by preserved copepods; and group D fed preserved copepods solely. Survival and biometric data were analysed together with major molecular biomarkers involved in growth, lipid metabolism and appetite. Moreover, fatty acid content of prey and larvae was also analysed. At the end of 40 days treatment, a stress test, on the remaining larvae, was performed to evaluate the effects of different diets on stress response. Data obtained evidenced a positive effect of cofeeding preserved copepods during sea bream larviculture. Higher survival and growth were achieved in group B (fed combined diet) larvae respect to control. In addition, preserved copepods cofeeding was able to positively modulate genes involved in fish growth, lipid metabolism, stress response and appetite regulation.

The role of dietary protein source on growth performance and lipid metabolism in European sea bass (Dicentrarchus labrax) was investigated, by the evaluation of fat content in different tissues, plasma metabolites, liver lipogenic activity and fatty acid composition of dorsal muscle tissue.Five isonitrogenous (490gkg-1crude protein dry-matter basis) and isolipidic (170gkg-1lipid dry-matter basis) diets with graded levels of wheat gluten, soybean and pea meal in substitution of fish meal were evaluated in European sea bass (initial body weight 23.9±0.1) for 96days. There was a significant difference (p

Copepods are the natural food items for marine fish larvae but are still difficult to be cultured on a continuous basis. Therefore, suitable storage techniques for copepod subitaneous eggs are of great interest. Cold-stored copepod embryos still develop and retain a physiological activity during the cold storage period. As a consequence, their nutritional quality may change over the time of storage with these changes possibly affecting both eggs viability and larval fish survival and development. In this study, the clownfish, Amphiprion polymnus, was used as an experimental model to compare, for the first time, the effects of diets based on enriched rotifers and Artemia (control), Acartia tonsa copepods of a continuous culture and A. tonsa originated from 6 months cold-stored eggs. The effects of the different diets were tested through morphometric, molecular and biochemical approach. This study demonstrated that after a 6 months cold storage period, the quality of copepods obtained from those eggs was suboptimal for A. polymnus larval rearing. In fact, larvae fed those copepods showed lower growth and survival performances respect to the other experimental groups.