Sustainable fish feeds with insects and probiotics positively affect freshwater and marine fish gut microbiota.

Aquaculture is the fastest-growing agricultural industry in the world. Fishmeal is an essential component of commercial fish diets, but its long-term sustainability is a concern. Therefore, it is important to find alternatives to fishmeal that have a similar nutritional value and, at the same time, are affordable and readily available. The search for high-quality alternatives to fishmeal and fish oil has interested researchers worldwide. Over the past 20 years, different insect meals have been studied as a potential alternate source of fishmeal in aquafeeds. On the other hand, probiotics—live microbial strains—are being used as dietary supplements and showing beneficial effects on fish growth and health status. Fish gut microbiota plays a significant role in nutrition metabolism, which affects a number of other physiological functions, including fish growth and development, immune regulation, and pathogen resistance. One of the key reasons for studying fish gut microbiota is the possibility to modify microbial communities that inhabit the intestine to benefit host growth and health. The development of DNA sequencing technologies and advanced bioinformatics tools has made metagenomic analysis a feasible method for researching gut microbes. In this review, we analyze and summarize the current knowledge provided by studies of our research group on using insect meal and probiotic supplements in aquafeed formulations and their effects on different fish gut microbiota. We also highlight future research directions to make insect meals a key source of proteins for sustainable aquaculture and explore the challenges associated with the use of probiotics. Insect meals and probiotics will undoubtedly have a positive effect on the long-term sustainability and profitability of aquaculture

Chronic social isolation affects feeding behavior of juvenile zebrafish (Danio rerio)

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

Wide-targeted gene expression infers tissue-specific molecular signatures of lipid metabolism in fed and fasted fish

European sea bass (Dicentrarchus labrax) is one of the most important species for Mediterranean aquaculture. It is therefore crucial to establish appropriate feeding management regimens and to gain better knowledge of nutritional requirements for this species, exploring not only new feed ingredients and feeding strategies, but also understanding the molecular mechanisms that regulate the metabolism of nutrients. Accordingly, transcriptomic analysis represents a useful nutrigenomic discovery tool for identifying the molecular basis of biological responses to nutrition as well as nutritional biomarkers in fish. This study evaluated how the transcriptional activity of genes controlling lipid metabolism in European sea bass were modulated in a tissue-specific manner in response to fasting and refeeding. Such approach focused on a panel of 29 genes in which desaturases, elongases, triacylglycerol lipases, fatty acid-binding proteins, β-oxidation and oxidative phosphorylation enzymes, phospholipid-related enzymes, and transcription factors that regulate lipid homeostasis were represented. Fasting activated the lipolytic machinery in adipose tissue, liver and muscle of European sea bass, whereas markers of lipogenesis were downregulated in liver and adipose tissue. Genes involved in phospholipid and oxidative metabolism were differentially regulated in liver and skeletal muscle of fasted European sea bass. However, 12 days of refeeding were sufficient, for the most part, to reverse the expression of key genes. Overall, our data clearly showed a tissue-specific regulation of lipid-related genes according to the different metabolic capabilities of each tissue, being the brain the most refractory organ to changes in nutrient and energy availability and liver the most responsive tissue.This research was partly funded by AQUAEXCEL EU 7 FP Project (Grant Agreement 262336): Trans National Access Grant to S.R. for accessing to IATS-CSIC facilities. This work has been partly funded under the EU seventh Framework Program by the ARRAINA Project N288925: Advanced Research Initiatives for Nutrition & Aquaculture.Peer Reviewe

The effects of dietary insect meal from Hermetia illucens prepupae on autochthonous gut microbiota of rainbow trout (Oncorhynchus mykiss)

This study evaluated the effects of dietary insect meal from Hermetia illucens larvae on autochthonous gut microbiota of rainbow trout (Oncorhynchus mykiss). Three diets, with increasing levels of insect meal inclusion (10%, 20%, and 30%) and a control diet without insect meal were tested in a 12-week feeding trial. To analyze the resident intestinal microbial communities, the Illumina MiSeq platform for sequencing of 16S rRNA gene and QIIME pipeline were used. The number of reads taxonomically classified according to the Greengenes database was 1,514,155. Seventy-four Operational Taxonomic Units (OTUs) at 97% identity were identified. The core of adhered intestinal microbiota, i.e., OTUs present in at least 80% of mucosal samples and shared regardless of the diet, was constituted by three OTUs assigned to Propiobacterinae, Shewanella, and Mycoplasma genera, respectively. Fish fed the insect-based diets showed higher bacterial diversity with a reduction in Proteobacteria in comparison to fish fed the fishmeal diet. Insect-meal inclusion in the diet increased the gut abundance of Mycoplasma, which was attributed the ability to produce lactic and acetic acid as final products of its fermentation. We believe that the observed variations on the autochthonous intestinal microbiota composition of trout are principally due to the prebiotic properties of fermentable chitin

Chitinase and insect meal in aquaculture nutrition: a comprehensive overview of the latest achievements.

The aquaculture industry is looking for sustainable alternatives to conventional fish meals in fish feed, and insect-based meals are proving to be a promising solution. These meals are nutritionally optimal as they have a high protein content and an ideal amino acid profile. However, the presence of chitin, a component of the insect exoskeleton in these meals presents both an opportunity and a challenge. Chitosan, a derivative of chitin, is known to improve the physiological functions of fish, including growth, immunity, and disease resistance. While chitin and its derivative chitosan offer several physiological benefits, their presence can affect the digestibility of feed in some fish species, making the inclusion of insect-based meals in aquafeeds complex. While studies suggest positive effects, some problems, such as reduced growth rates in certain species, emphasize the need for further research on chitin digestion in fish. Chitinase, an enzyme that breaks down chitin, is being investigated as a potential solution to improve the nutritional value of insect meals in aquafeed. This review provides a comprehensive analysis of the applications, benefits, and challenges of using chitinase in aquaculture, highlighting the enzyme’s role in improving feed digestibility, disease control, and environmental sustainability. Extensive research is required to fully understand the potential of chitinase enzymes in aquaculture and to optimize their applications in this dynamic field. Overall, this review provides insight into the evolving landscape of insect-based meals and the applications of chitinase enzymes within sustainable aquaculture practices

Next generation sequencing for gut microbiome characterization in rainbow trout (Oncorhynchus mykiss) fed animal by-product meals as an alternative to fishmeal protein sources

Animal by-product meals from the rendering industry could provide a sustainable and commercially viable alternative to fishmeal (FM) in aquaculture, as they are rich in most essential amino acids and contain important amounts of water-soluble proteins that improve feed digestibility and palatability. Among them, poultry by-product meal (PBM) have given encouraging results in rainbow trout (Oncorhynchus mykiss). However, the introduction of new ingredients in the diet needs to be carefully evaluated since diet is one of the main factors affecting the gut microbiota, which is a complex community that contributes to host metabolism, nutrition, growth, and disease resistance. Accordingly, we investigated the effects of partial replacement of dietary FM with a mix of animal by-product meals and plant proteins on intestinal microbiota composition of rainbow trout in relation to growth and feeding efficiency parameters. We used 1540 trout with an initial mean body weight of 94.6 +/- 14.2 g. Fish were fed for 12 weeks with 7 different feed formulations. The growth data showed that trout fed on diets rich in animal by-product meals grew as well as fish fed on control diet, which was rich in FM (37.3%) and PBM-free. High-throughput 16S rRNA gene amplicon sequencing (MiSeq platform, Illumina) was utilised to study the gut microbial community profile. After discarding Cyanobacteria (class Chloroplast) and mitochondria reads a total of 2,701,274 of reads taxonomically classified, corresponding to a mean of 96,474 +/- 68,056 reads per sample, were obtained. Five thousand three hundred ninety-nine operational taxonomic units (OTUs) were identified, which predominantly mapped to the phyla of Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria. The ratio between vegetable and animal proteins proved to play a central role in determining microbiome profiles and Firmicutes and Proteobacteria phyla were particularly discriminatory for diet type in trout. Plant ingredients favoured a higher Firmicutes: Proteobacteria ratio than animal proteins. Acceptable abundance of Firmicutes was guaranteed by including at least 25% of vegetable proteins in the diet regardless of animal protein source and percentage. In summary animal by-product meals, as replacements to FM, gave good results in terms of growth performances and did not induce significant changes in gut microbial richness, thus proving to be a suitable protein source for use in rainbow trout aqua feed