Insect-based diets high in lauric acid reduce liver lipids in freshwater Atlantic salmon

We evaluated the effect of a diet containing insect meal and insect oil on nutrient utilization, tissue fatty acid profile and lipid metabolism of freshwater Atlantic salmon (Salmo salar). Insect meal and insect oil from black soldier fly larvae (Hermetia illucens, L.; BSF), naturally high in lauric acid (12:0), were used to produce five experimental diets for an eight‐week feeding trial. 85% of the dietary protein was replaced by insect meal and/or all the vegetable oil was replaced by one of two types of insect oil. A typical industrial diet, with protein from fishmeal and soy protein concentrate (50:50) and lipids from fish oil and vegetable oil (33:66), was fed to a control group. The dietary BSF larvae did not modify feed intake or whole body lipid content. Despite the high content of saturated fatty acids in the insect‐based diets, the apparent digestibility coefficients of all fatty acids were high. There was a decrease in liver triacylglycerols of salmon fed the insect‐based diets compared to the fish fed the control diet. This is likely due to the rapid oxidation and low deposition of the medium‐chain fatty acid lauric acid.publishedVersio

Assessing amino acid solubility of black soldier fly larvae meal in Atlantic salmon (Salmo salar) in vivo and in vitro

In vitro and in vivo methods were used to evaluate amino acids solubility of black soldier fly (BSF) larvae meal and two experimental diets (reference and test diets) for Atlantic salmon. The current study used in vitro method such as pH stat to compare and standardise the salmon extracted enzyme (SE), and commercial enzyme (CE) based on their hydrolytic capacity on a purified protein substrate. Further, an in vitro amino acid solubility of feed ingredients and diets were measured using the standardised enzyme volume from SE and CE. Results showed that SE and CE exhibit similar protein hydrolytic capacity upon standardisation on purified substrates. However, when using the two-stage hydrolysis (acidic and alkaline steps), significantly higher amino acid solubility was observed with CE except for glycine, and proline which were equally solubilised by both SE, and CE. No significant difference was observed between reference and test diet using the SE except for tyrosine, valine, leucine, and phenylalanine, which were significantly higher solubilised in reference diet than test diet. Whereas higher solubility of valine, isoleucine, aspartic acid, and glutamic acid was observed in test diet using CE than SE. Similarly, the solubility of valine, isoleucine, and glutamic acid were higher in BSF larvae meal when CE was used. The in vivo true protein digestibility of BSF larvae meal was 99%, and 81% for the test diet containing BSF larvae meal. The results demonstrated a positive correlation (r = 0.91; p < 0.01) between salmon and commercial enzymes but overall, no significant correlation was observed for amino acid solubility between in vivo and in vitro. However, there was a strong positive correlation for protein solubility using SE (r = 0.98) than CE (r = 0.74) with the in vivo true protein digestibility. The efficiency of SE, and CE can be compared, and standardised based on DH%, and hence correlates better with the in vivo protein digestibility but not with amino acid solubilities.publishedVersio

The chemical composition of two seaweed flies (Coelopa frigida and Coelopa pilipes) reared in the laboratory

Two species of seaweed flies, Coelopa frigida and Coelopa pilipes, were reared in the laboratory and their larvae were sampled for composition of amino acids, fatty acids and elements. The larvae were grown on two different species of seaweed, Laminaria digitata and Fucus serratus. The aim was to gain knowledge on the influence of feeding media on the growth and composition of the larvae. Fucus serratus was more nutrient-dense than L. digitata, being richer in both protein and lipids, and thus led to ~70 % higher larvae growth. The larvae grown on F. serratus also had higher lipid and protein content than the larvae grown on L. digitata; F. serratus-grown larvae had ~8-9 % protein and ~18 % lipid (total fatty acids) (both values of dry matter), while the larvae grown on L. digitata had only ~7.5 % protein and ~13 % lipids. All seaweed flies had a similar and balanced amino acid composition, suitable for animal and human nutrition. The fatty acid composition was not highly affected by either insect species or feeding media, with all groups containing high concentrations of the monounsaturated fatty acid, palmitoleic acid (16:1n-7). The larvae also contained some fatty acids characteristic of marine environments, like eicosapentaenoic acid (20:5n-3), likely originating from the seaweed. Both species of seaweed fly larvae accumulated As, Cd, and Pb, but not Hg. The elevated levels of As and Cd in the larvae (highest measured concentrations 18.4 and 11.6 mg/kg, respectively, based on 12% moisture content) could potentially limit the use of seaweed fly larvae as a feed ingredient

A rapid acid hydrolysis method for the determination of chitin in fish feed supplemented with black soldier fly (Hermetia illucens) larvae

Insects are a natural source of feed for fish and have received more attention as a potential source of sustainable high-quality protein. However, contrasting results in different feeding trials have been ascribed to the chitin contained in the exoskeleton of insects and highlighted the importance of developing reliable methods for the quantification of chitin to draw meaningful conclusions about its effect on fish health. A rapid method based on the hydrolysis of chitin into glucosamine and further quantification by liquid chromatography tandem mass spectrometry is evaluated. The method offers good selectivity, linearity, limit of detection (1.08 × 10−5 % w/v or 5.38 × 10−4 % w/w), limit of quantification (3.26 × 10−5 % w/v or 1.63 × 10−3 % w/w), trueness (88.39–109.29 %) and precision (2.24–10.72 %). The quantitative method was successfully applied to real samples of fish feed supplemented with chitin from black soldier fly (Hermetia illucens) larvae.publishedVersio

Zinc uptake in fish intestinal epithelial model RTgutGC: Impact of media ion composition and methionine chelation

Apical uptake of zinc as ionic Zn(II) or as Zn-methionine (Zn-Met) was studied in RTgutGC cell line in vitro under media compositions mirroring the gut luminal ionic concentration of freshwater (FW) and seawater (SW) acclimated salmonids. Viability of the RTgutGC cells exposed to experimental media preparations showed a time-dependent decrease in SW treated cells, with the effect being significant at 48 h (P < 0.01), but not at 12 h or 24 h. Half effective concentration of Zn exposure over 12 h (EC50, in μM) was not differentially affected by media composition (FW, 59.7 ± 12.1 or SW, 83.2 ± 7.2; mean ± SE, P = 0.43). Zinc (65Zn) influx in RTgutGC was not different between FW or SW treated cells, but increased significantly in the presence of methionine (2 mM, L-Met or DL-Met). An interaction effect was observed between Zn concentration and media ionic composition on the impact of Met on apical Zn uptake (L-met, P < 0.001; DL-met, P = 0.02). In the presence of Met, apical Zn uptake in SW medium was significantly lower compared to FW, but only at higher Zn concentrations (12 and 25 μM, P < 0.01). Further, Met facilitated Zn uptake was reduced in cells treated with an amino acid transport system blocker with the effect being more significant and stereospecific in SW ionic conditions. The findings of this study showed that (i) Zn speciation in the presence of Met improved apical Zn uptake in RTgutGC cells and Zn-Met species were possibly taken up through Met uptake system. (ii) The effect was differentially affected by the ionic composition of the medium. Implications and limitations of the observations towards practical Zn nutrition of salmonids are discussed.publishedVersio

Application of laboratory methods for understanding fish responses to black soldier fly (Hermetia illucens) based diets

A major challenge for development of sustainable aquafeeds is its dependence on fish meal and fish oil. Replacement with more sustainable, nutritious and safe ingredients is now a priority. Over the last years, among several alternatives proposed, insects have received great attention as possible candidates. In particular, the black soldier fly (Hermetia illucens; BSF) represents a concrete example of how the circular economy concept can be applied to fish culture, providing a valuable biomass rich in fat and protein valorising organic by-products. In the last decade, several studies have been published about the use of different BSF dietary inclusion levels for various fish species including experimental models. Varying and encouraging results have been obtained in this research field using a plethora of laboratory methodological approaches that can be applied and coupled to obtain a comprehensive view of the BSF-based diets effects on fish physiology, health, and quality. The present review aims to explore some of the most promising laboratory approaches like histology, infrared spectroscopy, gut microbiome sequencing, molecular biology, fish fillets’ physico-chemical and sensory properties, essential for a better understanding of fish welfare and fillet quality, when BSF is used as aquafeed ingredient. In particular, great importance has been given to European finfish species and experimental models.publishedVersio

Potential of insect-based diets for Atlantic salmon (Salmo salar)

In the present study, we aimed to assess the effect of dietary insect meal (IM) and insect oil (IO) on growth performance, body composition and nutrient digestibility of freshwater reared Atlantic salmon. The IM and IO were produced from black soldier fly larvae (Hermetia illucens, L.; BSF) that had been grown on (1) media containing organic waste streams, or on (2) media partially containing seaweed (Ascophyllum nodosum). The feeding trial of the current study followed a factorial 2 × 3 way-ANOVA experimental design with six dietary groups of Atlantic salmon fed diets with insect-derived ingredients for 8 weeks. A typical industrial diet, with protein from fish meal and soy protein concentrate (SPC) (50:50) and lipids from fish oil and vegetable oil (33:66), was fed to a positive control group. Five experimental diets were formulated, where 85% of the dietary protein was replaced by IM and/or all the vegetable oil was replaced by IO (IM from insects grown on media 1, IO from insects grown on either media 1 (IO1) or media 2 (IO2)). Replacing the dietary fish meal and SPC with insect protein significantly reduced the apparent digestibility coefficients (ADC) of protein, lipid and all amino acids investigated, though remained highly digestible. There were, however, only small differences due to protein or lipid source in growth performance, and no effects of insect ingredients on feed intake or feed conversion ratio. Inclusion of IM-based diets significantly increased both hepatosomatic index and visceral somatic index of Atlantic salmon. Proteinase activity in the intestine was not affected by dietary inclusion of BSF larvae meal, while leucine aminopeptidase activity was lower in fish fed with insect ingredients than the control group. Whole-body protein, lipid, amino acids and minerals contents were not affected by protein or lipid source. In general, this study showed that protein meal and oil from BSF larvae hold a great potential as a source of nutrients for Atlantic salmon.publishedVersio

Changes in water temperature and oxygen: the effect of triploidy on performance and metabolism in large farmed Atlantic salmon

In salmon farming, the use of sterile triploids (3N) can mitigate the problem of escapees interbreeding with wild salmon. However, triploid salmon appear less tolerant to high water temperatures and low oxygen levels compared to diploids (2N). To investigate how the thermal performance and physiology of large (2.5 kg) triploid Atlantic salmon Salmo salar L. differs from those of diploids, both ploidies were subjected to water temperatures between 3 and 18°C. The fish were exposed to reduced oxygen saturations (O2 sat, 70%), termed hypoxia, at 6 and 18°C. Triploids fed more than diploids between 3 and 9°C and at similar levels at 12°C. At 15°C, the feed intake significantly dropped in both ploidies, although more in triploids. During hypoxia, feed intake was higher in triploids at 6°C and equal to diploids at 18°C. The overall feed conversion ratio was similar between ploidies. Muscle energy phosphates were generally lower in triploids than diploids, while muscle glucose, blood haemoglobin and haematocrit were lower in triploids than diploids at ≥12°C. Plasma lactate levels tended to be higher in triploids and increased with increasing temperature and at hypoxia in both ploidies. Plasma cortisol increased in both ploidies at high temperatures and was highest in triploids under hypoxic conditions at 18°C. Triploids had a higher cataract score at the start of the experiment and developed more cataracts throughout the experiment. The present findings show that large triploid Atlantic salmon perform better at colder water temperatures compared to diploids and differ in parts of their physiological expression at increasing and high temperature.publishedVersio