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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

There is scarce research on the use of black soldier fly larvae (BSFL) meal in the diets of Atlantic salmon (Salmo salar) reared in a real farm. Thus, the current study aimed to evaluate the effect of feeding BSFL meal to salmon in terms of fillet quality and sensory analysis at farm level conditions. Fish were fed a total of three diets (pellet size; 4.5 mm and 9 mm); a control diet (Control: 0% BSFL meal) formulated using a standard commercial recipe, containing FM (10%) and plant-based proteins. Two experimental diets were formulated using partially defatted BSFL meal (53% crude protein), in which plant-based proteins at 37% inclusion level from the control diet (4.5 mm pellets) were replaced by BSFL meal at 5% (BSFL 5%) and 10% (BSFL 10%). While for the 9.0 mm pellets, plant-based proteins at 35% were replaced by the BSFL meal at 5% and 10% inclusion level. All the three diets were formulated to be iso-nitrogenous and iso-lipidic diets. Diets were assigned to the sea-cages in triplicate (12 ×12 m; 1900 m3); housing ∼6000 fish per cage from 173 ± 2.7 g to harvest size 3.45 ± 74 kg for a period of 13 months. Feeding salmon with BSFL meal had no negative impact on the general fillet physical, chemical, and nutritional parameters. The fillet sensory evaluation showed that the control samples were associated with the typical attributes such as fresh salmon taste, while BSLF 10% to rancid taste but these attributes were not significantly different among the dietary groups. Overall, the current study showed that diet containing BSFL meal did not affect the general fillet parameters compared to salmon fed a commercial diet and thus validate the inclusion of BSFL meal up to 10% reared for a whole production cycle in sea cages.

The aims of the present work were to determine by meta-analysis the responses of broiler performance to increasing dietary levels of black soldier fly larvae meal (BSFLM) and to develop prediction models of the apparent metabolisable energy corrected to zero nitrogen balance (AMEn) of BSFLM. Eighteen studies from 12 countries that sum up to 20 experiments and involved a total of 63 treatments with 468 replicates and 4229 birds fulfilled the requirements for the meta-analysis. Four papers from three countries that investigated eight different samples of BSFLM were selected for development of AMEn prediction by means of regression analysis or artificial neural network (ANN) modelling. Growth performance results were not affected (p > .05) by the presence of BSFLM at levels of 5.7 ± 2.3 and 15.5 ± 6.0% in the diet. Multiple linear regression and ANN models showed high precision and accuracy, were unbiased and performed almost identically in the evaluation dataset (R2 = 0.856 and 0.858 and root mean square error of prediction as percentage of the observed mean = 5.216 and 5.241%). Feeding broilers with diets containing 15% BSFLM is not expected to have negative effects on growth performance. The following regression model could be useful for the estimation of AMEn in BSFLM: AMEn = 4.08 (± 0.330) + 0.295 (± 0.0158) × GE + 0.186 (± 0.0021) × EE; where AMEn and gross energy (GE) are MJ/kg dry matter and crude fat (EE) is % dry matter.

IntroductionThe 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.Materials and methodsTo 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 p 0.05). Despite the HH larvae being the most manipulated, no difference was also observed in final weight (HH, 160 mg; SH, 150 mg; p > 0.05) and survival rate (HH, 96.2%; SH, 94.5%; p > 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; p > 0.05). Finally, the development time from larva to fly (about 20.7; p > 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 (p > 0.05).DiscussionIn 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.

Abstract Background Sustainable aquaculture relies on multiple factors, including water quality, fish diets, and farmed fish. Replacing fishmeal (FM) with alternative protein sources is key for improving sustainability in aquaculture and promoting fish health. Indeed, great research efforts have been made to evaluate novel feed formulations, focusing especially on the effects on the fish gut microbiome. Few studies have explored host-environment interactions. In the present study, we evaluated the influence of novel insect-based (Tenebrio molitor) fish diets on the microbiome at the water-fish interface in an engineered rainbow trout (Oncorhynchus mykiss) farming ecosystem. Using 16S rRNA gene metabarcoding, we comprehensively analyzed the microbiomes of water, tank biofilm, fish intestinal mucus, fish cutis, and feed samples. Results Core microbiome analysis revealed the presence of a highly reduced core shared by all sample sources, constituted by Aeromonas spp., in both the control and novel feed test groups. Network analysis showed that samples were clustered based on the sample source, with no significant differences related to the feed formulation tested. Thus, the different diets did not seem to affect the environment (water and tank biofilm) and fish (cutis and intestinal mucus) microbiomes. To disentangle the contribution of feed at a finer scale, we performed a differential abundance analysis and observed differential enrichment/impoverishment in specific taxa, comparing the samples belonging to the control diet group and the insect-based diet group. Conclusions Omic exploration of the water-fish interface exposes patterns that are otherwise undetected. These data demonstrate a link between the environment and fish and show that subtle but significant differences are caused by feed composition. Thus, the research presented here is a step towards positively influencing the aquaculture environment and its microbiome.

Abstract Background The inclusion of alternative ingredients in poultry feed is foreseen to impact poultry gut microbiota. New feeding strategies (probiotics/prebiotics) must be adopted to allow sustainable productions. Therefore, the current study aimed to use metagenomics approaches to determine how dietary inclusion of prebiotic (inulin) plus a multi-strain probiotic mixture of Lactiplantibacillus plantarum and Lactiplantibacillus pentosus affected microbiota composition and functions of the gastro-intestinal tract of the broilers during production. Fecal samples were collected at the beginning of the trial and after 5, 11 and 32 days for metataxonomic analysis. At the end of the trial, broilers were submitted to anatomo-pathological investigations and caecal content was subjected to volatilome analysis and DNAseq. Results Probiotic plus prebiotic inclusion did not significantly influence bird performance and did not produce histopathological alterations or changes in blood measurements, which indicates that the probiotic did not impair the overall health status of the birds. The multi-strain probiotic plus inulin inclusion in broilers increased the abundance of Blautia, Faecalibacterium and Lachnospiraceae and as a consequence an increased level of butyric acid was observed. In addition, the administration of probiotics plus inulin modified the gut microbiota composition also at strain level since probiotics alone or in combination with inulin select specific Faecalibacterium prausnitzi strain populations. The metagenomic analysis showed in probiotic plus prebiotic fed broilers a higher number of genes required for branched-chain amino acid biosynthesis belonging to selected F. prausnitzi strains, which are crucial in increasing immune function resistance to pathogens. In the presence of the probiotic/prebiotic a reduction in the occurrence of antibiotic resistance genes belonging to aminoglycoside, beta-lactamase and lincosamide family was observed. Conclusions The positive microbiome modulation observed is particularly relevant, since the use of these alternative ingredients could promote a healthier status of the broiler’s gut.

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) but did reduce total n-3 fatty acids (P = 0.001) and increased total n-6 (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.

Sixteen 35-day-old male crossbred rabbits (New Zealand white × Thai native breed) with an initial weight of 484 ± 11.3 g were randomly divided into two groups of eight, constituting control and treatment groups. The treatment group was orally administered a crude extract of butterfly pea (Clitoria ternatea L.) at 0.5 g/kg body weight from weaning (at 35 days) to slaughter (at 90 days). The effects on the phagocytic activity of blood polymorphonuclear leukocytes, serum biochemistry, meat quality, muscular lipid peroxidation, the apparent digestibility of dry matter and nutrients, and gut histology were studied. The results revealed that the phagocytic function of circulating leukocytes (75 and 90 days) and alveolar macrophages (90 days) did not differ between the two groups. At slaughter, treated rabbits had lower blood urea nitrogen concentrations and higher liver weight than control rabbits (p < 0.05). After chilling at 4 °C for 24 h, a lower meat pH and the alteration of meat color (brighter, less yellow, lower hue angle, and decreased color saturation) were observed in the treated group (p < 0.05). Furthermore, lipid peroxidation (measured at 3, 5, and 7 storage days) in the meat of treated rabbits was lower than in controls (p < 0.05). The apparent digestibility of organic matter and ether extract (analyzed at 46 days for 4 days) was improved in the treated group (p < 0.05), whereas gut histology was unaffected. In conclusion, butterfly pea extract supplementation did not affect phagocytic function but led to a modification in meat color, delayed lipid peroxidation, and improved digestibility.

Due to the increasing costs of livestock farming, it is important to find cost-effective alternatives of feed stuffs. This study investigated the effects of high-moisture ear corn (HMEC) feeding on the production performance, serum antioxidant capacity, immunity, and ruminal fermentation and microbiome of dairy cows. Thirty pluriparous Chinese Holstein cows were randomly allocated to two groups: steam-flaked corn (SFC) and HMEC (replacement of 2 kg equal dry matter SFC) and fed for a 60 day trial. The results showed replacing SFC with HMEC significantly increased dry matter intake, milk yield, and 4% fat-corrected milk yield (p < 0.05). Serum levels of superoxide dismutase, glutathione peroxidase, and immunoglobulins G, M, and A were significantly higher, and those of creatinine and cholesterol were significantly lower, in the HMEC group than in the SFC group (p < 0.05). HMEC also significantly increased total volatile fatty acid and acetate (p < 0.05) concentrations. In both groups, the dominant phyla of ruminal bacteria were Bacteroidetes, Firmicutes, and Actinobacteria, and the dominant genera were Prevotella, NK4A214-group, and Succiniclasticum. Mogibacterium, Eubacterium nodatum group, norank-f-Lachnospiraceae, and Eubacterium brachy group were significantly enriched in the ruminal fluid of HMEC-group cows (p < 0.05). In conclusion, replacing SFC with HMEC improved production performance, antioxidant capacity, and immunity, while regulating both ruminal fermentation and the composition of the ruminal microbiome in dairy cows.

Abstract Background The most used protein sources in ruminant nutrition are considered as having negative impacts in terms of environmental sustainability and competition with human nutrition. Therefore, the investigation of alternative and sustainable feedstuffs is becoming a priority in ruminant production systems. Results This trial was designed to evaluate eight full-fat insect meals (Acheta domesticus – ACD; Alphitobius diaperinus – ALD; Blatta lateralis – BL; Gryllus bimaculatus – GB; Grylloides sygillatus – GS; Hermetia illucens – HI; Musca domestica – MD; and Tenebrio molitor – TM) as potential protein and lipid sources in ruminant nutrition. Fermentation parameters and fatty acids (FA) of rumen digesta after 24-h in vitro ruminal incubation of the tested insect meals were measured and compared with those of three plant-based meals (soybean meal, rapeseed meal and sunflower meal) and fishmeal (FM). Similarly to FM, the insect meals led to a significantly lower total gas production (on average, 1.75 vs. 4.64 mmol/g dry matter—DM), methane production (on average, 0.33 vs. 0.91 mmol/g DM), volatile FA production (on average, 4.12 vs. 7.53 mmol/g DM), and in vitro organic matter disappearance (on average, 0.32 vs. 0.59 g/g) than those observed for the plant meals. The insect meals also led to lower ammonia of rumen fluid, when expressed as a proportion of total N (on average, 0.74 vs. 0.52 for the plant and insect meals, respectively), which could be an advantage provided that intestinal digestibility is high. Differences in ruminal fermentation parameters between the insect meals could be partially explained by their chitin, crude protein and ether extract contents, as well as by their FA profile. In particular, high content of polyunsaturated FA, or C12:0 (in HI), seems to partially inhibit the ruminal fermentations. Conclusions The tested full-fat insect meals appear to be potentially an interesting protein and lipid source for ruminants, alternative to the less sustainable and commonly used ones of plant origin. The FA profile of the rumen digesta of ACD, ALD, GB, GS and TM, being rich in n-6 polyunsaturated FA, could be interesting to improve the quality of ruminant-derived food products.

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.

ABSTRACT: The development of a healthy gut during prestarter and starter phases is crucial to drive chicken's productivity. This study aimed to evaluate the effects of a thermomechanical, enzyme-facilitated, coprocessed yeast and soybean meal (pYSM) on growth performance, organ weights, leg health, and gut development in broiler chickens. A total of 576 as-hatched broiler chicks were randomly allotted to 3 dietary treatments (8 replicates/treatment, 24 chickens/replicate): a control group (C) without the pYSM, a treatment group 1 (T1), in which the pSYM was included at 20, 10, 5, 0, and 0% levels in the prestarter, starter, grower, finisher I, and finisher II feeding phases, respectively, and a treatment group 2 (T2), in which the pSYM was included at 5, 5, 5, 0, and 0% levels in each feeding phase. On d 3 and 10, 16 broilers/treatment were euthanized. The T1 broilers tended to show higher live weight (d 3 and 7) and average daily gain (prestarter and starter phases) than the other groups (P ≤ 0.10). Differently, pYSM-based diets did not influence the growth performance of the other feeding phases and the whole experimental period (P > 0.05). Relative weights of pancreas and liver were also unaffected by pYSM utilization (P > 0.05). Litter quality tended to have higher average scores in C group (P = 0.079), but no differences were observed for leg health (P > 0.05). Histomorphometry of gut, liver, and bursa of Fabricius was not affected by diet (P > 0.05). Gut immunity was driven to an anti-inflammatory pattern, with the reduction of IL-2, INF-γ, and TNF-α in the duodenum of treated birds (d 3, P < 0.05). Also, MUC-2 was greater in the duodenum of C and T2 group when compared to T1 (d 3, P = 0.016). Finally, T1-fed chickens displayed greater aminopeptidase activity in the duodenum (d 3 and 10, P < 0.05) and jejunum (d 3, P < 0.05). Feeding high levels of pYSM (10–20%) to broilers in the first 10 d tended to improve growth performance in the prestarter and starter phases. It also positively downregulated proinflammatory cytokines during the first 3 d, as well as stimulated the aminopeptidase activity in the prestarter and starter periods.

Abstract Herein, the effect of dietary inclusion of insect (Tenebrio molitor) meal on hepatic pathways of apoptosis and autophagy in three farmed fish species, gilthead seabream (Sparus aurata), European seabass (Dicentrarchus labrax) and rainbow trout (Oncorhynchus mykiss), fed diets at 25%, 50% and 60% insect meal inclusion levels respectively, was investigated. Hepatic proteome was examined by liver protein profiles from the three fish species, obtained by two-dimensional gel electrophoresis. Although cellular stress was evident in the three teleost species following insect meal, inclusion by T. molitor, D. labrax and O. mykiss suppressed apoptosis through induction of hepatic autophagy, while in S. aurata both cellular procedures were activated. Protein abundance showed that a total of 30, 81 and 74 spots were altered significantly in seabream, European seabass and rainbow trout, respectively. Insect meal inclusion resulted in individual protein abundance changes, with less number of proteins altered in gilthead seabream compared to European seabass and rainbow trout. This is the first study demonstrating that insect meal in fish diets is causing changes in liver protein abundances. However, a species-specific response both in the above mentioned bioindicators, indicates the need to strategically manage fish meal replacement in fish diets per species.

Abstract 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

The use of insect live larvae as environmental enrichment has recently been proposed in broiler chickens, but the concomitant administration of black soldier fly (BSF) and yellow mealworm (YM) has never been tested yet. Therefore, the present study aims to evaluate the effects of live BSF and YM larvae as environmental enrichments for broiler chickens by means of plumage status, behaviour, leg health, and excreta corticosterone metabolites (CM). A total of 180 4-day old male Ross 308 broiler chickens were randomly distributed in 3 experimental treatments (6 replicates/treatment, 10 birds/replicate) and fed for 35 days as follows: 1) control (C, commercial feed), 2) BSF: C + 5% of the expected daily feed intake [DFI] live BSF larvae and 3) YM: C + 5% of the expected DFI live YM larvae. Feathering, hock burn (HB) and footpad dermatitis (FPD) scores (end of the trial), as well as behavioural observations (beginning of the trial [T0] and every 11 days [T1, T2 and T3] during morning, larvae intake and afternoon) through video recordings, were assessed, and excreta samples collected to evaluate the CM. Feathering, HB and FPD scores, and excreta CM were unaffected by insect live larvae administration (p > 0.05). In the morning, the insect-fed birds displayed higher stretching, wing flapping, ground pecking (at T1 and T3), as well as lower preening (at T1 and T2), than the C group (p < 0.05). During the larvae intake, higher scratching, wing flapping and ground pecking, as well as lower stretching, preening and laying down, were observed in the insect-fed (scratching, stretching and laying down) or YM-fed (wing flapping, ground pecking and preening) groups than the C birds (p < 0.05). In the afternoon, insect live larvae administration increased wing flapping (YM) and laying down (BSF and YM), as well as decreased ground pecking (YM, p < 0.05). In conclusion, the administration of insect live larvae as environmental enrichment (especially YM) was capable of positively influencing the bird welfare through the stimulation of foraging behaviour, increase in activity levels, and reduction in bird frustration, without affecting the plumage status, leg health, and excreta CM.

Abstract Background The black soldier fly (Hermetia illucens) is the most promising insect candidate for nutrient-recycling through bioconversion of organic waste into biomass, thereby improving sustainability of protein supplies for animal feed and facilitating transition to a circular economy. Contrary to conventional livestock, genetic resources of farmed insects remain poorly characterised. We present the first comprehensive population genetic characterisation of H. illucens. Based on 15 novel microsatellite markers, we genotyped and analysed 2862 individuals from 150 wild and captive populations originating from 57 countries on seven subcontinents. Results We identified 16 well-distinguished genetic clusters indicating substantial global population structure. The data revealed genetic hotspots in central South America and successive northwards range expansions within the indigenous ranges of the Americas. Colonisations and naturalisations of largely unique genetic profiles occurred on all non-native continents, either preceded by demographically independent founder events from various single sources or involving admixture scenarios. A decisive primarily admixed Polynesian bridgehead population serially colonised the entire Australasian region and its secondarily admixed descendants successively mediated invasions into Africa and Europe. Conversely, captive populations from several continents traced back to a single North American origin and exhibit considerably reduced genetic diversity, although some farmed strains carry distinct genetic signatures. We highlight genetic footprints characteristic of progressing domestication due to increasing socio-economic importance of H. illucens, and ongoing introgression between domesticated strains globally traded for large-scale farming and wild populations in some regions. Conclusions We document the dynamic population genetic history of a cosmopolitan dipteran of South American origin shaped by striking geographic patterns. These reflect both ancient dispersal routes, and stochastic and heterogeneous anthropogenic introductions during the last century leading to pronounced diversification of worldwide structure of H. illucens. Upon the recent advent of its agronomic commercialisation, however, current human-mediated translocations of the black soldier fly largely involve genetically highly uniform domesticated strains, which meanwhile threaten the genetic integrity of differentiated unique local resources through introgression. Our in-depth reconstruction of the contemporary and historical demographic trajectories of H. illucens emphasises benchmarking potential for applied future research on this emerging model of the prospering insect-livestock sector.

Abstract 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.

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.

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.

The aquaculture industry is one of the fastest-growing sectors in animal food production. However, farming of carnivorous fish strongly relies on the use of wild fish-based meals, a practice that is environmentally and economically unsustainable. Insect-based diets constitute a strong candidate for fishmeal substitution, due to their high nutritional value and low environmental footprint. Nevertheless, data on the impact of insect meal (IM) on the gut microbiome of farmed fish are so far inconclusive, and very scarce in what concerns modulation of microbial-mediated functions. Here we use high-throughput 16S rRNA gene amplicon sequencing and quantitative PCR to evaluate the impact of different IMs on the composition and chitinolytic potential of the European sea bass gut digesta- and mucosa-associated communities. Our results show that insect-based diets of distinct origins differently impact the gut microbiota of the European sea bass (Dicentrarchus labrax). We detected clear modulatory effects of IM on the gut microbiota, which were more pronounced in the digesta, where communities differed considerably among the diets tested. Major community shifts were associated with the use of black soldier fly larvae (Hermetia illucens, HM) and pupal exuviae (HEM) feeds and were characterized by an increase in the relative abundance of the Firmicutes families Bacillaceae, Enterococcaceae, and Lachnospiraceae and the Actinobacteria family Actinomycetaceae, which all include taxa considered beneficial for fish health. Modulation of the digesta community by HEM was characterized by a sharp increase in Paenibacillus and a decrease of several Gammaproteobacteria and Bacteroidota members. In turn, a mealworm larvae-based diet (Tenebrio molitor, TM) had only a modest impact on microbiota composition. Further, using quantitative PCR, we demonstrate that shifts induced by HEM were accompanied by an increase in copy number of chitinase ChiA-encoding genes, predominantly originating from Paenibacillus species with effective chitinolytic activity. Our study reveals an HEM-driven increase in chitin-degrading taxa and associated chitinolytic activity, uncovering potential benefits of adopting exuviae-supplemented diets, a waste product of insect rearing, as a functional ingredient.

Abstract Background Aquaculture must continue to reduce dependence on fishmeal (FM) and fishoil in feeds to ensure sustainable sector growth. Therefore, the use of novel aquaculture feed ingredients is growing. In this regard, insects can represent a new world of sustainable and protein-rich ingredients for farmed fish feeds. Accordingly, we investigated the effects of full replacement of FM with Tenebrio molitor (TM) larvae meal in the diet of rainbow trout (Oncorhynchus mykiss) on fish gut and skin microbiota. Methods A feeding trial was conducted with 126 trout of about 80 g mean initial weight that were fed for 22 weeks with two isonitrogenous, isolipidic, and isoenergetic extruded experimental diets. Partially defatted TM meal was included in one of the diets to replace 100% (TM 100) of FM, whereas the other diet (TM 0) was without TM. To analyse the microbial communities, the Illumina MiSeq platform for sequencing of 16S rRNA gene and Qiime pipeline were used to identify bacteria in the gut and skin mucosa, and in the diets. Results The data showed no major effects of full FM substitution with TM meal on bacterial species richness and diversity in both, gut mucosa- and skin mucus-associated microbiome. Skin microbiome was dominated by phylum Proteobacteria and especially by Gammaproteobacteria class that constituted approximately half of the bacterial taxa found. The two dietary fish groups did not display distinctive features, except for a decrease in the relative abundance of Deefgea genus (family Neisseriaceae) in trout fed with insect meal. The metagenomic analysis of the gut mucosa indicated that Tenericutes was the most abundant phylum, regardless of the diet. Specifically, within this phylum, the Mollicutes, mainly represented by Mycoplasmataceae family, were the dominant class. However, we observed only a weak dietary modulation of intestinal bacterial communities. The only changes due to full FM replacement with TM meal were a decreased number of Proteobacteria and a reduced number of taxa assigned to Ruminococcaceae and Neisseriaceae families. Conclusions The data demonstrated that TM larvae meal is a valid alternative animal protein to replace FM in the aquafeeds. Only slight gut and skin microbiota changes occurred in rainbow trout after total FM replacement with insect meal. The mapping of the trout skin microbiota represents a novel contribution of the present study. Indeed, in contrast to the increasing knowledge on gut microbiota, the skin microbiota of major farmed fish species remains largely unmapped but it deserves thorough consideration.

The effects of Tenebrio molitor larvae meal (TM) dietary inclusion in rainbow trout diet were evaluated on muscular growth using gene expression and histomorphological features of liver, spleen, kidney, anterior and posterior gut through histopathological analyses. Two hundred fifty-two grow-out rainbow trout were fed four experimental diets containing increasing levels of TM: 0% (TM0), 5% (TM25), 10% (TM50), and 20% (TM100) corresponding to different levels of fish meal replacement (0, 25, 50, and 100%, respectively). Muscular growth was evaluated analysing the expression of various genes involved in different steps of myogenesis. Among the analysed genes, only MyoD expression resulted significantly higher in fish fed TM100 compared to fish fed TM0. The gut histomorphology was not affected by TM dietary inclusion and villus height differs from anterior and posterior segments regardless of the fed diet. Histopathological alterations were observed in all the sampled organs for all the dietary treatments; however, dietary TM inclusion did not influence either development or severity of the observed histopathological changes. The results obtained confirmed the safe utilisation of TM as an alternative protein source in rainbow trout diets and highlighted the necessity to deepen the studies of TM effect on the myogenesis process, especially at a molecular level.Highlights Rainbow trout can effectively be fed with a TM protein source. Total FM substitution by TM (TM100) decreased MyoD gene expression. Increasing TM dietary inclusion did not influence gut histomorphology.

This paper reviews current knowledge on two feedstuffs, that is, insect meal and fish by-products, as alternatives to conventional animal protein sources. After an introductory part that highlights the need for sustainable development of animal production, the alternative protein sources are discussed. In particular, after providing some indications on their production and supply focussing on EU, a SWOT (Strengths, Weaknesses, Opportunities, and Threats) analysis was performed to identify the key factors that could help or impair the development of both protein sources production sectors. Finally, future perspectives are presented. The use of processed animal proteins derived from insects in farmed fish feeding is recognised by the EU legislation that authorises the use of proteins from seven insect species and the allowed substrates to rear insects. Insects have several advantages in nutritional value and the amino acid composition of their proteins generally meet animal requirements for good growth and health. The SWOT analysis indicated that insect meals can be considered as feed functional ingredients with beneficial properties that depend on the insect species, rearing system adopted, and the substrate used for their growth. Insects are expected to be increasingly used as a replacement for conventional animal-derived proteins, especially in aquafeeds. In the section regarding fishery and aquaculture by-products, the potential use of raw materials obtained during seafood processing is discussed. Peptides and amino acids recovered from as hydrolysed proteins can be used in animal feeds to partially substitute conventional protein feedstuffs thus providing nutrients, bioactive compounds and feed additives for animals. The SWOT analysis identified opportunities and weaknesses. Both the alternative protein sources are promising alternative feed ingredients for livestock production.Highlights The sustainable development of animal production sector needs alternative protein sources for feeds formulation. Insects and fishery- and aquaculture by-products represent optimal alternative protein sources. A SWOT analysis has identified the key factors for the development of both protein sources production sectors.

The expected future demand for highly nutrient animal food products will push the animal production system to search for new sources of high-quality protein feedstuffs. In this scenario, economic and environmental issues will have to be considered while reducing the competition with the plant-based human food chains. Legume grains and some oilseed cakes, by-products from the oil industry, are the main protein sources for ruminants and terrestrial monogastrics such as pigs and poultry. Their relevant role will hold in the next decades, but it is necessary to increase the diversification of sources that can be grown profitably throughout the world, including European countries. Microalgae are a promising source of protein and other nutrients for animal feeding. However, an amazing richness of biologically active substances makes these organisms very interesting as feed ingredients, as their role go far beyond the supply of nutrients. Due to the limited usage of microalgae as human foodstuffs or food ingredients, low competition between microalgae-based feed and food chains is predictable. This review aims to synthesise current knowledge on minor pulses and other protein-rich plant products and microalgae, as alternative ingredients to the conventional animal protein sources, focussing on their production, availability, and nutritional values. Points of strength, weakness, opportunity and threat related to the use of these protein sources in animal feeding are separately analysed through a SWOT approach to underlie future needs in terms of research and/or technological development that could help valorise these nutrient sources as feed ingredients.

The present study addresses the effects of dietary Tenebrio molitor (TM) larvae meal inclusion on cytoprotective, cell death pathways, antioxidant defence, and intermediate metabolism in the heart, muscle, and digestive tract of gilthead seabream (Sparus aurata) and European sea bass (Dicentrarchus labrax). Three experimental diets were formulated to contain 0%, 25%, or 50% inclusion TM levels. Heat Shock Proteins (HSPs) induction was apparent in both species’ muscle at 50% inclusion. Conversely, p44/42 Mitogen-Activated Protein Kinase (MAPK) activation was increased (p

Abstract Background The constant interaction between diet and intestinal barrier has a crucial role in determining gut health in pigs. Hermetia illucens (HI) meal (that represents a promising, alternative feed ingredient for production animals) has recently been demonstrated to influence colonic microbiota, bacterial metabolite profile and mucosal immune status of pigs, but no data about modulation of gut mucin dynamics are currently available. The present study evaluated the effects of dietary HI meal inclusion on the small intestinal mucin composition of piglets, as well as providing insights into the cecal microbiota and the mucosal infiltration with immune cells. Results A total of 48 weaned piglets were randomly allotted to 3 dietary treatments (control diet [C] and 5% or 10% HI meal inclusion [HI5 and HI10], with 4 replicate boxes/treatment and 4 animals/box) and slaughtered after 61 days of trial (3 animals/box, 12 piglets/diet). The cecal microbiota assessment by 16S rRNA amplicon based sequencing showed higher beta diversity in the piglets fed the HI-based diets than the C (P

Abstract Background The relationship between diet and intestinal microbiota and mucin composition appears to be fundamental for poultry gut health. The effects of insect meal (whose role as alternative feed ingredient is now well recognized) on gut microbiota and mucin composition have recently been reported in Tenebrio molitor-fed free-range and broiler chickens, but no data are currently available for Hermetia illucens (HI)-fed broilers. The present study evaluated the effects of dietary HI meal inclusion on cecal microbiota and intestinal mucin composition of broiler chickens. Results A total of 256 male broiler chickens were allotted to 4 dietary treatments (control diet [C] and 5%, 10% and 15% HI meal inclusion, with 8 replicate pens/treatment and 8 birds/pen) and slaughtered at 35 d of age (2 animals/pen, 16 birds/diet). The cecal microbiota assessment by 16S rRNA amplicon based sequencing showed lower alpha diversity in HI15 chickens (Shannon, P

This study aimed to evaluate the effects of Lactobacillus acidophilus D2/CSL (L-1 × 109 cfu/kg feed/day) on biochemical parameters, faecal score (FS), cecal pH, gut morphometry, microbiota and cecal short-chain fatty acid (SCFAs) in rabbits. Three zootechnical trials were performed and in each trial 30 rabbits were allotted to two groups; a probiotic group (L) and a control group (C). At slaughter (day 45), samples of blood, duodenum, jejunum, ileum, liver and spleen were collected and submitted to histomorphometric analyses. Blood biochemical analyses, cecal microbiota and SCFAs determination were also performed. In trial 1 and 3, L. acidophilus D2/CSL did not affect productive parameters (p > 0.05). However, L group of trial 1 showed a lower morbidity and mortality compared to the control. In trial 2, C group showed a higher daily feed intake (p = 0.018) and a positive statistical tendency for live weight and average daily gain (p = 0.068). On the contrary, albumin was higher and ALFA-1 globulin was lower in the C group compared to L (p < 0.05). In all the trials, FS, cecal pH, histomorphometry, microbiota and SCFAs were unaffected. In conclusion, L. acidophilus D2/CSL did not impair growth performances, gut and rabbit’s health, reducing morbidity and mortality.

This study aimed to assess the effect of Hermetia illucens meal (HM) dietary inclusion on meagre oxidative status. Thus, fish were fed a fishmeal-based diet (CTR diet) and three other diets with increasing levels of HM inclusion, namely 10%, 20%, and 30% (diets HM10, HM20, and HM30, respectively). At the end of the trial, hepatic and intestine superoxide dismutase, catalase, and glucose-6-phosphate dehydrogenase activities and malondialdehyde concentration were unaffected by the diet composition. Liver glutathione peroxidase activity was higher in the fish fed the HM20 diet than in the fish fed the CTR and HM30 diets, and glutathione reductase activity linearly increased with the dietary HM level. The hepatic total glutathione and reduced glutathione contents were significantly lower in fish fed the HM20 diet than in fish fed the CTR and HM10 diets. In the intestine, the oxidized glutathione (GSSG) content and oxidative stress index linearly increased with the increase in dietary HM level, with the GSSG content of fish fed the HM20 diet being significantly higher than of fish fed the CTR diet. In conclusion, 30% HM might be included in meagre diets without negatively affecting hepatic and intestine oxidative status.

Effects of feeding dietary defatted black soldier fly (Hermetia illucens) larvae meal (HI) on intestine microbiota, and on histomorphology, oxidative enzyme activities in liver and intestine of pikeperch (Sander lucioperca) were investigated. Four isoproteic (45% crude protein) and isolipidic (18% ether extract) diets were formulated to include 0% (CO), 9% (HI9), 18% (HI18) and 36% (HI36) of HI as replacement for fishmeal at 0, 25, 50, and 100%, respectively, and were fed to triplicate groups of juvenile pikeperch (initial body weight, 68.7 ± 7.1 g) for 84 days. No adverse effects were detected on the intestine of pikeperch fed diet groups, in terms of histomorphology (P > 0.05), while fish fed free or low levels of HI (≤ 9% in diet) showed significant liver degeneration (P < 0.05). Dietary HI significantly affected the oxidative enzyme activities of catalase and glutathione peroxidase in the liver, and glutathione S-transferase in the intestine (P < 0.05), while activity of superoxide dismutase in both liver and intestine was HI-dose independent (P > 0.05). Feeding HI-containing diets positively modulated the richness and diversity of intestinal microbiota, especially for HI18 group (P < 0.05). Inclusion HI up to 18% (50% fishmeal replacement) in pikeperch diets increased abundance of Clostridium, Oceanobacillus, Bacteroides, and Faecalibacterium genera, whereas the predominant bacterium, Cetobacterium was found in control and HI36 groups. This study reveals the potential of HI as an immune and health booster for juvenile pikeperch.

Abstract 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.

Abstract Background The aim of this trial was to investigate the effects of different inclusion levels of a partially defatted black soldier fly (BSF, Hermetia illucens L.) larva meal on the growth performance, nutrient digestibility, blood profile, gut morphology and histological features of piglets. A total of 48 newly weaned piglets were individually weighed (initial body weight (IBW): 6.1 ± 0.16 kg) and randomly allocated to 3 dietary treatments (4 boxes as replicates/treatment and 4 animals/box). BSF larva meal was included at increasing levels (0% [BSF0], 5% [BSF5] and 10% [BSF10]) in isonitrogenous and isoenergetic diets formulated for two feeding phases: I (from d 1 to d 23) and II (from d 24 to d 61). The weight gain (WG), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) were calculated for each feeding phase and for the whole trial. The haematochemical parameters and nutrient digestibility of the piglets were also evaluated. A total of 3 piglets per box were slaughtered on d 61 and the slaughtered piglets were submitted to morphometric investigations and histopathological examinations. Results No overall significant differences were observed for growth performance (P > 0.05), except for the ADFI of phase II, which showed a linear response to increasing BSF meal levels (P 0.05). Conclusions The obtained results show that a partially defatted BSF larva meal can be used as a feed ingredient in diets for weaned piglets without negatively affecting their growth performance, nutrient digestibility, blood profile, gut morphology or histological features.

Volatile fatty acid production (VFA) in the caeca and fatty acid profile of meat and fat depots were investigated on 62 days old broiler equally divided in 2 groups fed with a corn-soybean (SBM) or a corn-insect meal (TML, from Tenebrio molitor larvae) based diet from 30 days of age. The total amount of VFAs was higher (p

The feed utilisation of young chicks is characterised by a suboptimal fat digestibility, which can be improved by means of dietary emulsifiers. The objective of this study was to evaluate the effect of dietary Globin on the energy efficiency and digestibility of starter feeds and on the production performance of broilers throughout the whole rearing cycle. A total of 224-day-old ROSS 708 chickens (14 birds/pen, 8 replicates/treatment) were fed ad libitum with either a basal diet (C) or a basal diet with the addition of 0.05% Globin during the starter (d1–10), growing (d10–25) and finisher (d25–35) periods. Nutrient digestibility (aD), protein metabolisability (aMCP), energy efficiency (EE) and net energy for production (NEp) were assessed during the starter period. The average daily feed intake (ADFI), average daily gain (ADG), feed conversion ratio (FCR) and protein efficiency ratio (PER) were measured of each growth period. Globin significantly decreased FCR (p = .020) and increased aDfat (p = .021), EE (p = .028) and NEp (p = .011) during the starter period. aMCP (p = .049) and PER (p = .039) were higher in the Globin group than in the Control group. The increased availability of energy from dietary fat, as a result of Globin supplementation, possibly shifted the use of the absorbed amino acids towards an anabolic metabolism, and this could explain the increased aMCP and PER but similar aDCP. The overall performance was similar between groups, although Globin tended to increase PER (p = .064) overall.Highlights Globin dietary supplementation was studied in broiler chickens. Globin improved digestibility and nutrient efficiency utilization in the first period. The overall performance was similar between groups.

Abstract Background The present work aimed at evaluating the effect of the dietary replacement of soybean oil (S) by two types of insect fats extracted from black soldier fly larvae (H, Hermetia illucens L.) and yellow mealworm larvae (T, Tenebrio molitor L.) on growth performance, nutrient digestibility, blood parameters, intestinal morphology and health of growing rabbits. Methods At weaning, 200 crossbred rabbits (36 days old) were allotted to five dietary treatments (40 rabbits/group): a control diet (C) containing 1.5% of soybean oil and four experimental diets where soybean oil was partially (50%) or totally (100%) substituted by H (H50 and H100) or T (T50 and T100) fats. Total tract digestibility was evaluated on 12 rabbits per treatment. The growth trial lasted 41 d and, at slaughtering (78 days old), blood samples were collected from 15 rabbits per treatment, morphometric analyses were performed on duodenum, jejunum and ileum mucosa, and samples of liver, spleen and kidney were submitted to histological evaluation. Results No difference was observed between the control and the experimental groups fed insect fats in terms of performance, morbidity, mortality and blood variables. The addition of H and T fats did not influence apparent digestibility coefficients of dry matter, protein, ether extract, fibre fractions and gross energy. Gut morphometric indices and organ histopathology were not affected by dietary inclusion of H and T fats. Conclusions H and T fats are suitable sources of lipid in rabbit diets to replace soybean oil without any detrimental effect on growth performance, apparent digestibility, gut mucosa traits and health.

Insect meal (IM), recently authorized for use in aquafeeds, positions itself as a promising commodity for aquafeed inclusion. However, insects are also rich in chitin, a structural polysaccharide present in the exoskeleton, which is not digested by fish, resulting in lower fish performance. Through the application of a dietary pressure, this study aimed to modulate European sea bass gut microbiota towards the enrichment of chitinolytic bacteria to allow the isolation of novel probiotics capable of improving the use of IM-containing diets, overcoming chitin drawbacks. Five isoproteic (44%) and isolipidic (18%) diets were used: a fish meal (FM)-based diet (diet CTR), a chitin-supplemented diet (diet CHIT5), and three diets with either 25% of Hermetia illucens and Tenebrio molitor larvae meals (HM25 and TM25, respectively) or H. illucens exuviae meal (diet HEM25) as partial FM substitutes. After an 8-week feeding trial, the results showed a clear modulatory effect towards spore-forming bacteria by HM25 and HEM25 diets, with the latter being responsible for the majority of the chitinolytic fish isolates (FIs) obtained. Sequential evaluation of the FI hemolytic activity, antibiotic resistance, total chitinolytic activity, sporulation, and survival in gastrointestinal-like conditions identified FI645 and FI658 as the most promising chitinolytic probiotics for in vivo application.

Abstract Background Gut health in poultry depends on the balance between the host, intestinal microbiota, intestinal microscopic features and diet. The effects of insect meal (a promising alternative protein source for poultry feed) on chicken gut morphology have recently been reported, but no data about intestinal microbiota and mucin composition modulation are available. The present study evaluated the effects of dietary Tenebrio molitor (TM) meal inclusion on gut health of free-range chickens by intestinal microbiota, morphology and mucin composition characterization. Results One hundred forty female medium-growing hybrids were divided into 2 dietary treatments (control feed [C] and 7.5% TM inclusion, with 5 replicate pens/treatment and 14 birds/pen) and slaughtered at 97 days of age (2 birds/pen for a total of 10 chickens/diet). The gut microbiota assessment on cecal content samples by 16S rRNA amplicon based sequencing showed higher alpha (Shannon, P

The expected global population growth to 9.7 billion people in 2050 and the significant change in global dietary patterns require an increase in global food production by about 60%. The protein supply for feed and food is most critical and requires an extension in protein sources. Edible insects can upgrade low-grade side streams of food production into high-quality protein, amino acids and vitamins in a very efficient way. Insects are considered to be the “missing link” in the food chain of a circular and sustainable economy. Insects and insect-derived products have entered the European market since first being acknowledged as a valuable protein source for feed and food production in around 2010. However, today, scaling up the insect value chain in Europe is progressing at a relatively slow pace. The mission of SUSINCHAIN (SUStainable INsect CHAIN)—a four-year project which has received funding from the European Commission—is to contribute to novel protein provision for feed and food in Europe by overcoming the remaining barriers for increasing the economic viability of the insect value chain and opening markets by combining forces in a comprehensive multi-actor consortium. The overall project objective is to test, pilot and demonstrate recently developed technologies, products and processes, to realize a shift up to Technology Readiness Level 6 or higher. In addition to these crucial activities, the project engages with stakeholders in the insect protein supply chain for feed and food by living labs and workshops. These actions provide the necessary knowledge and data for actors in the insect value chain to decrease the cost price of insect products, process insects more efficiently and market insect protein applications in animal feed and regular human diets that are safe and sustainable. This paves the way for further upscaling and commercialization of the European insect sector.

The slaughter performance and meat quality of two native Italian chicken breeds, Bionda Piemontese (BP, n = 64) and Bianca di Saluzzo (BS, n = 64), were investigated. Two-way ANOVA, considering breed, sex, and their interaction, was used to compare the properties of birds slaughtered at 5, 6, 7, and 8 months of age. Subsequently, data were analyzed using one-way ANOVA and the Duncan test to evaluate the differences between slaughter ages. The BP breed produced a better carcass yield than BS at 5, 7, and 8 months of age (p < 0.05). Breast moisture and crude protein contents were influenced by gender, and were higher in males than in females (p < 0.05). By contrast, the crude fat content was higher in females than in males (p < 0.05). The saturated fatty acid content of breast meat increased as the birds aged in both breeds (p < 0.05). The polyunsaturated fatty acid content of both breast and thigh meat was higher in males than in females (p < 0.001 and p < 0.05, respectively). In general, slaughtering at 7 months was associated with the best slaughter and meat quality characteristics in both breeds. Moreover, from a nutritional point of view, the characteristics of the meat from male birds were preferable to those of meat from females.

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.

Abstract Background The present study has evaluated the effects of different inclusion levels of a partially defatted black soldier fly (Hermetia illucens L.; HI) larva meal on the growth performance, blood parameters and gut morphology of broiler chickens. A total of 256 male broiler chickens (Ross 308) were reared from d 1 to d 35 and assigned to 4 dietary treatments (8 replicates/treatment and 8 birds/replicate). HI larva meal was included at increasing levels (0, 5%, 10% and 15%; HI0, HI5, HI10 and HI15, respectively) in isonitrogenous and isoenergetic diets formulated for 3 feeding phases: starter (1–10 d), growing (10–24 d) and finisher (24–35 d). Two birds per pen were slaughtered at d 35 and morphometric investigations and histopathological alterations were performed. Results The live weight (LW) showed linear and quadratic responses to increasing HI larva meal (maximum for HI10 group). Average daily gain (ADG) showed a linear and quadratic responses to HI meal (maximum for HI10 group) during starter and growing periods. A linear decrease was observed for ADG during the finisher period. The daily feed intake (DFI) showed a linear and quadratic effect during the starter period (maximum for HI10 group). Linear and quadratic responses were observed for the feed conversion ratio (FCR) in the growing period and for the whole period of the experiment. The FCR showed a linear response in the finisher period (maximum for HI15). No significant effects were observed for the blood and serum parameters, except for the phosphorus concentration, which showed linear and quadratic responses as well as glutathione peroxidase (GPx) activity, the latter of which showed a linear response. The HI15 birds showed a lower villus height, a higher crypt depth and a lower villus height-to-crypt depth ratio than the other groups. Conclusions Increasing levels of dietary HI meal inclusion in male broiler chickens may improve the LW and DFI during the starter period, but may also negatively affect the FCR and gut morphology, thus suggesting that low levels may be more suitable. However, no significant effects on the haematochemical parameters or histological findings were observed in relation to HI meal utilization.

Objective The present experiment has tested the effect of dietary alfalfa flavonoids (AAF) supplementation on the productive performances, carcass characteristics, meat quality and lipid oxidation of growing rabbits. Methods One hundred and sixty crossbred rabbits (42 days old) were divided into four groups of forty animals each and were fed either a control diet (AAF0) or an AAF0 diet supplemented with 400, 800, or 1,200 mg of AAF/kg per diet (AAF4, AAF8, and AAF12, respectively) from weaning to slaughtering (102 days old). Performance data were recorded over a period of 60 days. At the end of the trial, 12 rabbits were slaughtered per group, and the carcass characteristics were recorded. Moreover, the plasma, liver and dorsal muscles were sampled from 12 rabbits/group, and were analyzed for lipid oxidation. Results No significant differences were recorded for the performance, carcass characteristics and meat quality traits except for lightness parameter that was lower in the control group. Dietary AAF supplementation significantly (p

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.

In tropical and subtropical regions, as well as in the internal and/or marginal Mediterranean areas, one of the most important problems related to animal production is represented by the inadequate nutritional supplies. The low productivity of the animals, often connected to reduced annual growth, is, in fact, not infrequently attributable to the low nitrogen content and the high fiber content of the local plant species and crop residues that constitute the base ingredients of the rations commonly adopted by farmers. The use of the supplementation with arboreal and shrub fodder, although often containing anti-nutritional factors and toxins that limit its use, could be a profitable way to alleviate the nutritional deficiencies of the basic diets. Leucaena leucocephala (Lam.) De Wit is native to Central America and widely naturalized in the majority of Latin American countries. It is a legume suitable for tropical and subtropical environments including the countries of the Mediterranean area. Moreover, its spread is desirable if we consider the multiple uses to which it is suitable, the considerable amount of biomass produced, and its role in preserving the environment. The aim of this work was to highlight the characteristics of Leucaena that can justify its wide diffusion. A structured analysis of strengths and weaknesses was performed accordingly. Being a good protein source for feeding livestock, it could be a species to be introduced in the inland areas of the Mediterranean countries as an alternative protein source; the limit represented by the presence of anti-nutritional factors could be overcome by feed processing and by launching targeted research programs.

In this study, a total of 200 male broiler chickens (Ross 308) were assigned to four dietary treatments (5 pens/treatment and 10 birds/pen) for two feeding phases: starter (0–11 days of age) and grower-finisher (11–33 days of age). A basal diet containing soy oil (SO) as added fat was used as control group (C), tested against three experimental diets where the SO was partially substituted by BSF larvae fat (BSF) or one of two types of modified BSF larvae fat (MBSF1 and MBSF2, respectively). The two modified BSF larvae fats had a high and low ratio of monobutyrin to monoglycerides of medium chain fatty acid, respectively. Diet did not influence the growth or slaughter performance, pH, color, or the chemical composition of breast and thigh muscles, gut morphometric indices, or histopathological alterations in all the organs. As far as fecal microbiota are concerned, MBSF1 and MBSF2 diets reduced the presence of Clostridium and Corynebacterium, which can frequently cause infection in poultry. In conclusion, modified BSF larva fat may positively modulate the fecal microbiota of broiler chickens without influencing the growth performance and intestinal morphology or showing any adverse histopathological alternations.

The inclusion of insects as a protein source in feed production is not only related to technical, economical, and normative restrictions but is also affected by consumer acceptance. In this study, we evaluated consumers’ attitudes, intention to purchase and eat, and willingness to pay for meat obtained from a farmed duck fed with insect-based meal or a live insect diet. We conducted a survey among a sample of 565 consumers to test the effects of information about the benefits of using insects as feed on consumers’ attitudes towards animal-based products fed with insects. Providing information on the sustainability and nutritional benefits of using insects as feed increased both attitude towards and intention to purchase and eat meat products made from animals fed with insects. In the treatment group, we found a significant reduction from 21.9 to 14.0% in those who wanted to be compensated for buying a duck fed with an insect-based meal and an increase in those willing to pay the same price—from 64.9 to 72.7%. The information treatment significantly increased the intention to eat such products, suggesting that increasing consumers’ knowledge might help in reducing the fears and misconceptions around the topic of using insects as a feed source.

Metabolic rates are linked to the energetic costs of different activities of an animal’s life. However, measuring the metabolic rate in free-swimming fish remains challenging due to the lack of possibilities to perform these direct measurements in the field. Thus, the calibration of acoustic transmitters with the oxygen consumption rate (MO2) could be promising to counter these limitations. In this study, rainbow trout (Oncorhynchus mykiss Walbaum, 1792; n = 40) were challenged in a critical swimming test (Ucrit) to (1) obtain insights about the aerobic and anaerobic metabolism throughout electromyograms; and (2) calibrate acoustic transmitters’ signal with the MO2 to be later used as a proxy of energetic costs. After this calibration, the fish (n = 12) were implanted with the transmitter and were followed during ~50 days in an aquaculture facility, as a case study, to evaluate the potential of such calibration. Accelerometer data gathered from tags over a long time period were converted to estimate the MO2. The MO2 values indicated that all fish were reared under conditions that did not impact their health and welfare. In addition, a diurnal pattern with higher MO2 was observed for the majority of implanted trout. In conclusion, this study provides (1) biological information about the muscular activation pattern of both red and white muscle; and (2) useful tools to estimate the energetic costs in free-ranging rainbow trout. The use of acoustic transmitters calibrated with MO2, as a proxy of energy expenditure, could be promising for welfare assessment in the aquaculture industry.

The aim of the present study was to evaluate the effects of dietary bilberry pomace (BP) on colour, cooking losses, proximate composition and fatty acid (FA) profile of hind leg meat in growing rabbits. One hundred and forty-four Grimaud weaned rabbits (35 days old) were randomly divided into four groups of 36 animals each and fed ad libitum with a basal diet (C diet) tested against three assay diets developed by substituting 50, 100 and 150 g/kg of the C diet with BP (BP5, BP10 and BP15 diets, respectively). At 83 days of age, the rabbits were slaughtered without fasting. Inclusion of BP in the diet did not significantly affect colour, cooking losses, ash and protein contents, but determined a significant increase of ether extract in the muscle. Increasing dietary inclusion of BP also determined a proportional increase of total polyunsaturated fatty acids (PUFA) and total n-3 FA, as well as a proportional decrease of total saturated fatty acids (SFA), total branched chain fatty acids (BCFA) and total monounsaturated fatty acids (MUFA) in the muscle. Dietary BP significantly improved the PUFA/SFA ratio, the Σ n-6/Σ n-3 FA ratio and the atherogenicity, thrombogenicity and hypocholesterolemic/hypercholesterolemic FA indexes of rabbit meat but it decreased Δ5-desaturase plus Δ6-desaturase index. The obtained results suggest that BP inclusion in growing rabbit diets can improve the fatty acid profile of hind leg meat, with consequent health benefits to consumers.

Abstract Background The study aimed to determine the apparent total tract digestibility coefficients (ATTDC) of nutrients, the apparent metabolizable energy (AME and AMEn) and the amino acid (AA) apparent ileal digestibility coefficients (AIDC) of a partially defatted (BSFp) and a highly defatted (BSFh) black soldier fly larvae meal. The experimental diets were: a basal diet and two diets prepared by substituting 250 g/kg (w/w) of the basal diet with BSFp or BSFh, respectively. Results Significant differences were found between BSFp and BSFh meals for ATTDC of the nutrients: BSFp resulted more digestible than BSFh, except for ATTDC of CP which did not differed between meals, while a statistical trend was observed for ATTDC of DM and EE. The AME and AMEn values were significantly (P

The present research studied for the first time the potential application of the fat derived from the black soldier fly larvae fat (BSLF) in substitution to the soybean oil in the diet for broiler chickens: growth performances, feed-choice, blood traits, carcass characteristics and meat quality were considered in this study. A total of 150 male broiler chicks (Ross 308) at one-day of age were randomly allotted to 3 dietary treatments (5 replicates and 10 birds/pen): a basal control diet (C group), and the same diet in which the soybean oil was replaced by 50% (CH group) or 100% (H group) BSLF. Growth performances, feed-choice test, blood traits and slaughtering performances were not influenced by diets. Independently of BSLF inclusion, broiler chickens breast meat had also similar crude protein and ether extract contents and displayed similar thawing loss. Furthermore, pH, L*, a*, b* colour values, and drip loss were unaffected by dietary treatments both at 0 and 9 days of refrigerated storage. As expected, the fatty acid profile of broiler chickens breast was greatly affected by BSLF inclusion level. With increasing BSLF inclusion rate, the proportion of SFA increased (32.2, 37.8, 43.5% for C, CH and H breast meat, respectively, p