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 ± 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-α, and tlr-5) was observed in medium intestine in the H25 and H50 groups (p < 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.

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Intensive fish farming relies on the use of feeds based on fish meal and oil as optimal ingredients; however, further development of the aquaculture sector needs new, nutritious and sustainable ingredients. According to the concept of circular economy, insects represent good candidates as aquafeed ingredients since they can be cultured through environmental-friendly, cost-effective farming processes, on by-products/wastes, and many studies have recently been published about their inclusion in fish feed. However, information about the physiological effects of insect-based diets over the whole life cycle of fish is presently missing. At this regard, the present study investigated, for the first time, the effects of Black Soldier Fly based diets (25 and 50% fish meal substitution) administration for a six months period in zebrafish ( Danio rerio ), from larvae to adults. A multidisciplinary approach, including biometric, biochemical, histological, spectroscopic and molecular analyses was applied. Aside a general reduction in fish growth and lipid steatosis, six-months feeding on Black Soldier Fly based diets did not show major negative effects on zebrafish. Gut histological analysis on intestine samples did not show signs of inflammation and both stress markers and immune response markers did not show significant differences among the experimental groups.

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. The promotion of high-quality fish production as well as fish welfare are the main aquaculture goals, both strictly related to an adequate fish nutrition 1. The use of fish meal (FM) and fish oil (FO) in aquafeed formulation is no more feasible because of important environmental and economic issues 2. Over the last decades, several alternative ingredients (plant origin proteins, microalgae and processed animal proteins) to FM and FO have been tested 3. However, each of these ingredients showed some disadvantages in its application in aquafeed formulation including unbalanced amino acid profile, poor protein digestibility, presence of anti-nutritional factors and high production costs 3,4. Insects are now considered as an alternative and sustainable ingredient for feed production 5. In particular, the Black Soldier Fly (Hermetia illucens; BSF) larvae are one of the most promising candidates because of their proper protein content and the amino acid composition similar to that of FM 6,7. In addition, BSF have low environmental requirements, a high feed conversion efficiency and they can growth on organic by-products, promoting sustainability and the circular economy concept in the aquaculture sector 8,9. Several studies tested different BSF inclusion levels in aquafeed formulation but results on fish physiological responses are still controversial, while behavioural effects on fish are completely missing 10-13. The use of BSF in aquafeed has been shown, at certain inclusion levels,

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.

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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.