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.

Eight PM10 aerosol samples were collected in the vicinity of the "Mario Zucchelli" Italian Antarctic Station (formerly Terra Nova Bay Station) during the 2000-2001 austral summer using a high-volume sampler and precleaned cellulose filters. The aerosol mass was determined by differential weighing of filters carried out in a clean chemistry laboratory under controlled temperature and humidity. A two-step sequential extraction procedure was used to separate the water-soluble and the insoluble (dilute-HCl-extractable) fractions. Cd, Pb and Cu were determined in the two fractions using an ultrasensitive square wave anodic stripping voltammetric (SWASV) procedure set up for and applied to aerosol samples for the first time. Total extractable metals showed maxima at midsummer for Cd and Pb and a less clear trend for Cu. In particular, particulate metal concentrations ranged as follows: Cd 0.84-9.2 microg g(-1) (average 4.7 microg g(-1)), Pb 13.2-81 microg g(-1) (average 33 microg g(-1)), Cu 126-628 microg g(-1) (average 378 microg g(-1)). In terms of atmospheric concentration, the values were: Cd 0.55-6.3 pg m(-3) (average 3.4 pg m(-3)), Pb 8.7-48 pg m(-3) (average 24 pg m(-3)), Cu 75-365 pg m(-3) (average 266 pg m(-3)). At the beginning of the season the three metals appear widely distributed in the insoluble (HCl-extractable) fraction (higher proportions for Cd and Pb, 90-100%, and lower for Cu, 70-90%) with maxima in the second half of December. The soluble fraction then increases, and at the end of the season Cd and Pb are approximately equidistributed between the two fractions, while for Cu the soluble fraction reaches its maximum level of 36%. Practically negligible contributions are estimated for crustal and sea-spray sources. Low but significant volcanic contributions are estimated for Cd and Pb (approximately 10% and approximately 5%, respectively), while there is an evident although not quantified marine biogenic source, at least for Cd. The estimated natural contributions (possibly including the marine biogenic source) cannot account for the high fractions of the metal contents, particularly for Pb and Cu, and this suggests that pollution from long-range transport is the dominant source.

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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