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To maximise the black soldier fly (BSF) bioconversion capacity, a waste that is not suitable for use as it is, can have a positive effect if combined with others in a formulation. In this perspective, a total of 3600 of 6-day-old larvae were inoculated on 6 iso-proteic, iso-lipidic and iso-energetic diets (named from D1 to D6; 100 larvae/replicate; 6 replicates/treatment) formulated with 21 by-products. To evaluate growth performance, 30 larvae per replicate were randomly sampled every 4 days until the 40% reached the prepupae phase, in which survival rate (SR), larva-prepupae time duration and bioconversion indexes were calculated. When a pupa was identified, it was moved in the emergence box (23 pupae/treatment) to evaluate the adult parameters: pupa-fly duration time, fly live weight, weight reduction, fly lifespan, puparium weight and emergence rate (ER). Considering SR (>92%) and ER (>94.6%), all diets were suitable for the BSF breeding. The D1 and D5 (composed by brewers spent grain, vegetable and fruit waste) groups overall performed the best larvae growth and development time, while D2 (composed by glaze waste and buffalo milk whey) displayed the worst (p < 0.05). The D5 maintained good weight in prepupae, pupae and adult stages and also D2 reached the same outcome. The bioconversion indexes followed the trend of the larvae performance. In conclusion, the results herein obtained allowed to discriminate the protein, fat and energy variables on the BSF life history traits and observe other factors of influence, such as the ash content, which will need to be evaluated with future research.

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.

The black soldier fly is considered the most promising insect species for mass production; however, information on the effects of handling, which is unavoidable during experimental trials and rearing practices, is still limited. To address this gap, three different manipulation intensities were tested on 100 6-day-old larvae per replica (6 replicates/treatments) fed on Gainesville diet: (1) hard-handled (HH), larvae underwent continuous manipulation until the end of larval stage, (2) soft-handled (SH), larvae were manipulated after the appearance of the first prepupa, (3) no-handled (NH), larvae remained untouched. Every 4 days from the beginning to the end of the larval stage, the manipulations lasted 30 min and occurred under laboratory conditions (20°C). During the sampling operations, at least 30 larvae were randomly extracted, washed, dried, and weight-mimicked. At the end of larval stage, all the boxes remained untouched until the adult fly stage, and the emergency rate and sex ratio were evaluated on dead flies. Data were statistically analyzed using IBM SPSS V20.0.0 software and the considered significance level was  < 0.05. The larval stage lasted 8.2 days for both HH and SH (  > 0.05). Despite the HH larvae being the most manipulated, no difference was also observed in final weight (HH, 160 mg; SH, 150 mg;  > 0.05) and survival rate (HH, 96.2%; SH, 94.5%;  > 0.05). The manipulation did not influence the bioconversion capacity of the larvae (bioconversion efficiency corrected for the residue: HH, 14.3%; SH, 12.91%; reduction rate: HH, 58.4%; SH, 55.9%; waste reduction index: HH, 7.28%/day; SH, 7.25%/day;  > 0.05). Finally, the development time from larva to fly (about 20.7;  > 0.05), the emergency rate (NH: 92.8%; SH: 89.5%; HH: 82.7%) and sex ratio (~1.2% to male flies) were not affected by the handling (  > 0.05). In conclusion, the handling procedures used in the current study did not influence the life history traits of the black soldier fly. However, further studies are needed to evaluate if different experimental protocols on various scales, the colony strain or other handling procedures may suggest a different scenario or confirm the results.

This study evaluated the effects of dietary Hermetia illucens (HI) meal on growth performance, nutrient digestibility, general health status and gut health of post-weaning piglets reared under commercial conditions. A total of 336, 28–days-old mixed-sex weaned piglets were randomly allotted to 4 dietary treatments (control [C] and 3, 6 and 9% HI meal inclusion [HI3, HI6 and HI9], with 4 replicate boxes/treatment, 21 animals/box and 3 feeding phases) and slaughtered after 63 days of trial (4 animals/box, 16 piglets/diet). In the whole trial, the average daily gain and feed conversion ratio showed a linear response to dietary HI meal inclusion (p < 0.05). The nutrient digestibility also displayed a linear and/or quadratic response to dietary HI meal inclusion during phases I and II, while the opposite trend was highlighted during phase III (p < 0.05). Increased blood antioxidant response was also identified for HI6- and HI9-fed animals (p < 0.01). Dietary HI meal inclusion did not influence gut histomorphology or organ histopathological findings, as well as caecal microbiota alpha and beta diversity. However, selected amplicon sequence variants were associated with specific dietary treatments (p < 0.05), such as Dorea, Faecalitalea, Bacteroides, Anaerostipes and Oscillospiraceae (HI3), Phascolarctobacterium, Rhodospirillales and Odoribacter (HI9), Lachnospiraceae, Oscillospiraceae, Anaerovoracaceae, Faecalibacterium, Lachnospira, Alistipes, Lachnoclostridium and Acinetobacter (HI3 and HI6), and Enterobacter, Roseburia, Ruminococcaceae, Lachnospiraceae and Bifidobacterium (HI-based diets). In conclusion, including up to 9% of HI meal enhanced growth performance and antioxidant response, and partially improved nutrient digestibility and caecal microbiota of the weaned piglets, without impairing their gut morphology and overall health status.

Background The provision of environmental enrichments to Muscovy ducks could reduce the expression of the aggressive behaviors. The aim of the present study was to evaluate the effects of black soldier fly (BSF) and yellow mealworm (YM) live larva provision on Muscovy duck performance, excreta corticosterone metabolites (ECM), behavior, and blood parameters. Methods A total of 126 3-day-old female Muscovy ducklings were allotted to 18 pens (6 replicates/treatment, 7 birds/pen) and assigned to 3 experimental treatments: a control group fed commercial feed, and two experimental treatments fed commercial feed plus the 5% (based on the expected daily feed intake, as fed basis) of BSF and YM live larvae (BSF and YM groups, respectively). A two-phase feeding program was applied: starter (from 3 to 31 days of age) and grower-finisher (from 32 to 55 days of age). The live weight, average daily gain, average daily feed intake, and feed conversion ratio were calculated. Larva consumption times were collected, and video recordings were performed during 3 periods (P) each day: the hour before (P1), during (P2), and after (P3) the larva administration. ECM were evaluated at 3, 31, and 55-day-old. Finally, the total red and white blood cell counts, serum proteins, lipids, and liver and renal function serum enzymes were evaluated on 12 birds/treatment. Results The experimental treatment did not affect the growth performance of the birds ( P  > 0.05). Larva consumption times were always similar between the two insect species, except at 14–18 days of age, were BSF larvae were consumed faster than YM larvae ( P  < 0.001). The birds showed less walking activity during P2, and preening behavior increased in YM birds during P3. The C birds increased the attack behavior over the weeks ( P  < 0.05). During weeks 1–3 the YM group reduced the attack frequency (P1 > P3; P  < 0.05). Finally, the provision of live BSF and YM larvae significantly reduced the ECM at 55 days of age and the heterophil to lymphocyte ratio ( P  < 0.05). Conclusions Live BSF and YM larva supplementation in Muscovy duck improves duck welfare, without impairing birds’ growth performance.

This study focused on the effects of including a mixture of Hermetia illucens (HI) and Tenebrio molitor (TM) meals on feed colour, growth performance, carcass yield, somatic indexes, nutrient digestibility, intestinal microbiota, histomorphometry, and fillet quality of rainbow trout (Oncorhynchus mykiss). Seven diets were formulated: one control diet (CTRL) containing 15% fishmeal, two diets where the fishmeal was replaced with 100% HI (HI100) or TM (TM100) meal, and four diets containing increasing levels of a mix (1:1) of HI and TM meals, replacing 25% (MIX25), 50% (MIX50), 75% (MIX75) and 100% (MIX100) of the fishmeal. A total of 378 fish (average weight: 126 ± 1.71 g (g)) were randomly distributed in 21 experimental tanks (3 replicates/treatment and 18 fish/replicate) and fed for 84 days. A digestibility trial was also conducted to assess the apparent nutrient digestibility coefficients of the diets. Diet significantly affected pellet colour (L*, a*, b*; p < 0.001), with CTRL and MIX25 being the lightest and HI100 and MIX75 the darkest. Fillet colour, pH24h, and drip loss were significantly affected by diet (p < 0.001). The pH24h parameter, reflecting stress and freshness, was lowest in MIX25 and highest in MIX75, suggesting a synergistic effect and potential impact on shelf life. No significant differences were found in growth performance, carcass yield, somatic indexes, fillet chemical composition, intestinal microbiota, and the histopathology of internal organs among the experimental diets (p > 0.05), nor in nutrient digestibility. This study confirms that HI and TM meals, alone or combined, are viable alternatives in low-fishmeal diets for rainbow trout.

Background Black soldier fly (BSF) is one of the most promising species for the intensive breeding of insects given its adaptability and its efficiency in the conversion of waste. To maximize the production and use waste as substrates, it is essential to determine the larvae nutritional requirements. The study aims to evaluate the effects of 5 practical, semi-purified and isoenergetic diets (PSPID) with increasing protein levels (10%, CP10; 14%, CP14; 16%, CP16; 19%, CP19) on BSF life history traits. A total of 2000 six-day-old larvae were weighed and divided into groups of 100 (4 replicates/treatment [PSPID and Gainesville diet (GA) used as environmental control]). In larva-prepupa stage, sampling was carried out on 30 individuals/replicate, while in prepupa-adult stage on 35. Results In the whole larval stage, the CP16 treatment performed better when compared to the other groups. At 18 days old, the CP14 treatment showed a higher weight than the CP19 ( P  < 0.01), while the CP10 and CP16 weights were intermediate. On the contrary, the CP10 prepupae weight was higher than the CP19 ( P  < 0.01). The CP10 and CP14 pupae performed better in terms of weight when compared to the others ( P  < 0.01). The dietary treatments did not affect the adult parameters. The sex significantly influenced both the exuvia weight, which was the greatest in females ( P  < 0.001), and the fly lifespan, longest in males ( P  < 0.05). Fly lifespan was also influenced by the interaction between treatment and sex, with the CP19 females showing a longer life than the others ( P  < 0.05). Conclusions In conclusion, the optimal protein level considering the whole larval stage is 16% on dry matter (DM). At 18 days old, looking at the size intended for the meal production, it can be considered 14% on DM. The result obtained on adult emergence in PSPID may not be attributable to the protein content. Further researches on macronutrients requirements determination have to be conducted to evaluate the substrates composition effects on BSF life history traits.

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.

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.

HIGHLIGHTS The study highlights the feasibility of supplementing commercial feed for medium-growing chickens with live BSFL determined minimal modifications to growth performance and slaughter outcomes. The FCR of the chickens improved by live BSFL supplementation. The supplementation of the BSFL in diet affected the fatty acid composition of the meat without compromising its quality.