Defatted black soldier fly (Hermetia illucens) in pikeperch (Sander lucioperca) diets: Effects on growth performance, nutrient digestibility, fillet quality, economic and environmental sustainability.

The use of insect meal in aquafeed formulations has recently gained attention. Detailed knowledge about the inclusion levels for pikeperch (Sander lucioperca), a promising candidate for intensive aquaculture in Europe remains, however, fragmented. In the present study, 4 isoproteic (45% dry matter) and isoenergetic (21 MJ/kg) diets were formulated, including a control diet (H0) containing 30% fishmeal (FM) on an as-fed basis and the other 3 diets in which FM protein was replaced by defatted black soldier fly (Hemetia illucens) meal (HIM) at 25%, 50%, and 100% (diet abbreviation H9, H18 and H36, corresponding to an inclusion level of 9%, 18% and 36%, respectively). The feeding trial was performed in triplicate groups of 50 juvenile pikeperch (mean weight, 68.7 g) fed with experimental diets for 84 d during which the growth performance, nutrient digestibility, fillet quality and economic and environmental sustainability of rearing pikeperch were evaluated. Our findings indicated that pikeperch in H0, H9, and H18 groups displayed better results regarding growth performance indices, except for survival rate where no significant difference among groups was recorded (P = 0.642). A significantly lower organ-somatic index, including hepatosomatic, viscerosomatic and perivisceral fat index, was found in fish in H18 groups than other groups (P < 0.05). Inclusion of HIM affected the digestibility of the nutrients and resulted in an almost linear reduction in the apparent digestibility coefficient of dry matter and protein. Concerning the fillet quality, dietary HIM negatively affected the protein and ash contents of the fish fillets, while the crude fat remained unchanged. Dietary HIM did not significantly modify total saturated, monounsaturated and polyunsaturated fatty acids in the fillets of fed pikeperch (P > 0.05) but did reduce total n-3 fatty acids (P = 0.001) and increased total n-6 (P < 0.001). Increasing inclusion levels of HIM reduced the environmental impacts associated with fish in-to-fish out ratio but entailed heavy burdens on energy use and eutrophication. Low and moderate inclusion levels of HIM did not negatively affect land use and water use compared to an HIM-free diet (P > 0.05). The addition of HIM at a level as low as 9% elicited a similar carbon footprint to that of the control diet. The economic conversion ratio and economic profit index were negatively affected at increased insect meal inclusion levels. This study has shown that the incorporation of HIM in feed formulations for pikeperch is feasible at inclusion levels of 18% without adverse effects on growth performance parameters. The feasibility also highlighted the environmental benefits associated with land use and marine resources required to produce farmed fish.

How Different Dietary Methionine Sources Could Modulate the Hepatic Metabolism in Rainbow Trout?

In aquafeeds in which plant proteins are used to replace fishmeal, exogenous methionine (Met) sources are demanded to balance the amino acid composition of diets and meet the metabolic fish requirements. Nonetheless, since different synthetic Met sources are commercially available, it is important to determine their bioavailability and efficacy. To address this issue, we conducted a two-month feeding trial with rainbow trout (Oncorhynchus mykiss), which were fed diets supplemented with five different forms of Met: Met-Met, L-Met, HMTBa, DL-Met, and Co DL-Met. No differences in growth performance were found in trout fed with different Met forms, but changes in the whole-body composition were found. In particular, Met-Met and L-Met promoted a significant body lipid reduction, whereas the protein retention was significantly increased in fish fed with HMTBa and Co DL-Met. The latter affected the hepatic Met metabolism promoting the trans-sulfuration pathway through the upregulation of CBS gene expression. Similarly, the L-Met enhanced the remethylation pathway through an increase in BHMT gene expression to maintain the cellular demand for Met. Altogether, our findings suggest an optimal dietary intake of all tested Met sources with similar promoting effects on fish growth and hepatic Met metabolism. Nevertheless, the mechanisms underlying these effects warrant further investigation.

On the Compatibility of Fish Meal Replacements in Aquafeeds for Rainbow Trout. A Combined Metabolomic, Proteomic and Histological Study.

The sustainable development of modern aquaculture must rely on a significant reduction of the fish meal (FM) used in aquafeed formulations. However, FM substitution with alternative ingredients in diets for carnivorous fish species often showed reduced nutrient absorption, significantly perturbed metabolisms, and histological changes at both hepatic and intestinal levels. In the present study, rainbow trout (Oncorhynchus mykiss) were fed three different experimental aquafeeds. A control diet with higher FM content (27.3%) than two test formulations in which FM was substituted with two more sustainable and promising alternatives: insect meal (Hermetia illucens larvae = 10.1%, FM = 11.6%) and poultry by-products meal (PBM = 14.8%; FM = 11.7%). Combined metabolomics and proteomics analyses of fish liver, together with histological examination of liver and intestine demonstrated that a well-balanced formulation of nutrients in the three diets allowed high metabolic compatibility of either substitution, paving the way for a deeper understanding of the impact of novel raw materials for the fish feed industry. Results show that the main metabolic pathways of nutrient absorption and catabolism were essentially unaltered by alternative feed ingredients, and also histological alterations were negligible. It is demonstrated that the substitution of FM with sustainable alternatives does not have a negative impact on fish metabolism, as long as the nutritional requirements of rainbow trout are fulfilled.

Efficacy of Utilization of All-Plant-Based and Commercial Low-Fishmeal Feeds in Two Divergently Selected Strains of Rainbow Trout (Oncorhynchus mykiss): Focus on Growth Performance, Whole-Body Proximate Composition, and Intestinal Microbiome.

The present study aimed to investigate the growth performance, whole-body proximate composition, and intestinal microbiome of rainbow trout strains when selected and non-selected for weight gain on all-plant protein diets. A 2x2 factorial design was applied, where a selected (United States) and a non-selected (ITA) rainbow trout strain were fed using either an all-plant protein (PP) or a commercial low-FM diet (C). Diets were fed to five replicates of 20 (PP) or 25 (C) fish for 105 days. At the end of the trial, growth parameters were assessed, and whole fish (15 pools of three fish/diet) and gut samples (six fish/diet) were collected for whole-body proximate composition and gut microbiome analyses, respectively. Independent of the administered diet, the United States strain showed higher survival, final body weight, weight gain, and specific growth rate when compared to the ITA fish (p < 0.001). Furthermore, decreased whole-body ether extract content was identified in the PP-fed United States rainbow trout when compared to the ITA strain fed the same diet (p < 0.001). Gut microbiome analysis revealed the Cetobacterium probiotic-like genus as clearly associated with the United States rainbow trout, along with the up-regulation of the pathway involved in starch and sucrose metabolism. In summary, the overall improvement in growth performance and, to a lesser extent, whole-body proximate composition observed in the selected rainbow trout strain was accompanied by specific, positive modulation of the intestinal microbiome.

Modulation of Antioxidant Defense in Farmed Rainbow Trout (Oncorhynchus mykiss) Fed with a Diet Supplemented by the Waste Derived from the Supercritical Fluid Extraction of Basil (Ocimum basilicum).

Phytotherapy is based on the use of plants to prevent or treat human and animal diseases. Recently, the use of essential oils and polyphenol-enriched extracts is also rapidly increasing in the aquaculture sector as a means of greater industrial and environmental sustainability. Previous studies assessed the antibacterial and antiparasitic effects of these bioactive compounds on fish. However, studies on the modulation of oxidative stress biomarkers are still scant to date. Thus, in this study, the modulation of antioxidant defense against oxidative stress exerted by fish diets supplemented with a basil supercritical extract (F1-BEO) was assessed in rainbow trout Oncorhynchus mykiss. The F1-BEO extracted with supercritical fluid extraction was added to the commercial feed flour (0.5, 1, 2, 3% w/w) and mixed with fish oil to obtain a suitable compound for pellet preparation. Fish were fed for 30 days. The levels of stress biomarkers such as superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glyoxalase I, glyoxalase II, lactate dehydrogenase, glutathione and malondialdehyde showed a boost in the antioxidant pathway in fish fed with a 0.5% F1-BEO-supplemented diet. Higher F1-BEO supplementation led to a failure of activity of several enzymes and the depletion of glutathione levels. Malondialdehyde concentration suggests a sufficient oxidative stress defense against lipid peroxidation in all experimental groups, except for a 3% F1-BEO-supplemented diet (liver 168.87 ± 38.79 nmol/mg prot; kidney 146.86 ± 23.28 nmol/mg prot), compared to control (liver 127.76 ± 18.15 nmol/mg prot; kidney 98.68 ± 15.65 nmol/mg prot). Our results suggest supplementing F1-BEO in fish diets up to 0.5% to avoid potential oxidative pressure in farmed trout.

Effect of Insect Live Larvae as Environmental Enrichment on Poultry Gut Health: Gut Mucin Composition, Microbiota and Local Immune Response Evaluation.

The aim of this study was to evaluate the effect of Hermetia illucens (HI) and Tenebrio molitor (TM) live larvae as environmental enrichment on the mucin composition, local immune response and microbiota of broilers. A total of 180 four-day-old male broiler chickens (Ross 308) were randomly allotted to three dietary treatments (six replicates/treatment; ten animals/replicate): (i) control (C); (ii) C+HI; (iii) C+TM. Live larvae were distributed based on 5% of the expected daily feed intake. At slaughter (39 days of age), samples of duodenum, jejunum and ileum (twelve animals/diet) were submitted to mucin histochemical evaluation. Expression of MUC-2 and cytokines was evaluated by rt-qPCR in jejunum. Mucin staining intensity was not influenced by diet (p > 0.05); however, this varied depending on the intestinal segment (p < 0.001). No significant differences were recorded for IL-4, IL-6 TNF-α, MUC-2 and INF-γ gene expression in jejunum, while IL-2 was lower in the TM group compared to HI and C (p = 0.044). Caecal microbiota showed higher abundance of Clostridium, Saccharibacteria and Victivallaceae in the HI group, while Collinsella was higher in the TM group. The results suggested that live insect larvae did not impair mucin composition or local immune response, and can slightly improve caecal microbiota by enhancing a minor fraction of short chain fatty acid-producing taxa.

Dietary inclusion of a partially defatted black soldier fly (Hermetia illucens) larva meal in low fishmeal-based diets for rainbow trout (Oncorhynchus mykiss).

BACKGROUND: Recent investigations highlighted that Hermetia illucens (BSF) larva meal can be a valuable alternative protein source for aquafeed production. In this study, in substitution of fishmeal, we used increasing inclusion levels of a partially defatted BSF larva meal in low fishmeal-based diets for rainbow trout (Oncorhynchus mykiss Walbaum) and we evaluated the related implications in terms of growth performance, physical characteristics, proximate and fatty acid (FA) compositions of the fillets, gut and liver histology, and diet digestibility. In a 131-day trial, 576 fish (100.1 ± 9.29 g) were randomly allotted to 24 tanks (24 fish/tank, 4 replicates/treatment). Six experimental diets were produced to have partial replacement of fishmeal (0, 10%, 20%, 30%, 40% and 50%) by increasing levels of BSF meal (0% [BSF0], 3% [BSF3], 6% [BSF6], 9% [BSF9], 12% [BSF12] and 15% [BSF15] on as fed basis, respectively). RESULTS: No differences were observed among the treatments for all the considered growth performance parameters. The viscero and hepato-somatic indexes showed significant differences among the treatments, with the highest values observed in the BSF15 group. No differences were recorded in terms of fillet's physical characteristics, dry matter (DM), crude protein (CP) and ether extract (EE) contents. Total saturated and monounsaturated FA increased, while polyunsaturated FA (particularly n-3 FA) decreased while increasing the HI meal inclusion in the diet. Histopathology of liver and gut was not affected, whereas, in posterior gut, villi were higher in BSF6 and BSF9 compared to BSF3 fish. The apparent digestibility of DM, CP, EE and gross energy did not vary among the treatments. CONCLUSIONS: These results suggest that a partially defatted BSF meal could be included up to 15% in low fishmeal-based diets for rainbow trout with no adverse effects on growth performance, fillet's physical characteristics, gut and liver health, and diet digestibility. On the contrary, the fillet FA composition worsened while increasing the level of BSF meal in the diet.

Partially Defatted Tenebrio molitor Larva Meal in Diets for Grow-Out Rainbow Trout, Oncorhynchus mykiss (Walbaum): Effects on Growth Performance, Diet Digestibility and Metabolic Responses.

Insect meals are good candidates to replace fishmeal as new protein sources in aquafeeds. This study evaluated the effects of fishmeal replacement with different dietary inclusion levels of a partially defatted Tenebrio molitor (L.) larva meal (TM) on rainbow trout (Oncorhynchus mykiss Walbaum) growth, diet digestibility, and hepatic intermediary metabolism. A 154-day growth trial was performed with 252 rainbow trout (78.3 ± 6.24 g) randomly divided into twelve tanks and fed four experimental diets containing increasing levels of TM: 0% (TM0), 25% (TM25), 50% (TM50), and 100% (TM100) of fishmeal substitution, corresponding to TM dietary inclusion levels of 0%, 5%, 10%, and 20%, respectively. A digestibility trial was performed feeding 180 rainbow trout (94.6 ± 7.31 g) with the experimental diets used in the growth trial. The growth parameters were not affected by TM dietary inclusion. Regarding the evaluated apparent digestibility coefficients (ADC), only the ADC of crude protein was affected, showing the following trend: TM0 = TM25 > TM50 > TM100. The activities of key hepatic amino acid catabolic and lipogenic enzymes were not affected by the dietary composition. The results suggest that a partially defatted TM could totally replace fishmeal in commercial rainbow trout diets without negative effects on fish performance.

Could Dietary Black Soldier Fly Meal Inclusion Affect the Liver and Intestinal Histological Traits and the Oxidative Stress Biomarkers of Siberian Sturgeon (Acipenser baerii) Juveniles?

The trial investigates if a highly defatted Hermetia illucens larva meal (H) at two dietary inclusion levels and a vegetable protein based diet (VEG) influences the normal gut and liver histology and the oxidative stress biomarkers in liver and kidney of Siberian sturgeon juveniles. Fish were fed four diets: one control diet (H0) containing 70% of fishmeal (FM), two diets including 18.5% (H185) and 37.5% (H375) of highly defatted H in substitution for 25% and 50% of FM, and one vegetable protein based diet (VEG). At the end of a growth trial, 12 fish per treatment were sacrificed by over-anaesthesia to collect 12 liver and 5 distal intestine samples for histological analyses, as well as 12 liver and kidney samples for biochemical analyses. The H and VEG diets did not significantly affect the histology of liver and distal intestine, but alterations of the oxidative stress biomarkers were detected at the highest inclusion level of H (37.5%). In order to avoid unfavorable effects on the fish health, an inclusion level up to 18.5% of H is recommended for Siberian sturgeon juveniles.

Nutritional effects of the dietary inclusion of partially defatted Hermetia illucens larva meal in Muscovy duck.

BACKGROUND: The present work is aimed at evaluating the effect of different inclusion levels of a partially defatted black soldier fly (Hermetia illucens, L.; HI) larva meal for ducks. A total of 192 female 3-day-old Muscovy ducklings (Cairina moschata domestica, Canedins R71 L White, Grimaud Freres Selection, France) were divided into 4 groups, assigned 4 different dietary treatments (6 replicates/treatment and 8 birds/replicate) and reared from 3 to 50 days of age. HI larva meal was included at increasing levels (0, 3%, 6% and 9%, HI0, HI3, HI6 and HI9, respectively) in isonitrogenous and isoenergetic diets formulated for 3 feeding phases: starter (3-17 days of age), grower (18-38 days of age) and finisher (39-50 days of age). The growth performance and apparent total tract digestibility were evaluated during the trial using titanium dioxide as an inert marker (0.3% of inclusion). At 51 days of age, two birds per pen were slaughtered and histomorphological investigations were performed. RESULTS: The live weight and average daily gain showed a quadratic response to increasing HI meal in the grower period (minimum corresponding to the HI6 group). No effects of dietary inclusion levels were observed for the daily feed intake or feed conversion ratio. The apparent dry matter and organic matter digestibility were not affected by the dietary treatment. A linear decrease was observed for the crude protein apparent digestibility in the starter period (minimum for the HI9 groups). The ether extract apparent digestibility increased linearly during the grower and finisher periods (minimum for the HI0 group). The morphometric indices were not influenced by the dietary treatments. CONCLUSIONS: The inclusion of up to 9% of HI partially defatted larva meal in the diet of ducks did not cause any effect on growth performance, as well as the apparent digestibility. Moreover, dietary HI inclusion preserved the physiological intestinal development.