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Background: The gut microbiota of black soldier fly larvae (BSFL, Hermetia illucens) play a crucial role in recycling various organic waste streams. This capability is linked to the presence of a potential common core microbiota in BSFL. However, subjective thresholds for defining core taxa and the difficulty of separating genetic and environmental influences have prevented a clear consensus in the literature. We analysed the gut bacterial communities of two genetically distinct BSF lines (wild type (WT) and lab-adapted line (LD)) raised on ten different diets based on common agricultural by-products and food waste in Southeast Asia. Results: High-throughput 16S rRNA gene sequencing revealed that gut bacterial communities were significantly influenced by genetics (p = 0.001), diet (plant/meat-dominated; p = 0.001), larval age (p = 0.001), and the interactions between all three (p = 0.002). This led us to investigate both common core taxa and lineage-specific core taxa. At a strict > 97% prevalence threshold, four core taxa were identified: Providencia_A_732258, an unclassified genus within the family Enterococcaceae, Morganella, and Enterococcus_H_360604. A relaxed threshold (> 80% prevalence) extended the core to include other potential common core taxa such as Klebsiella, Proteus, and Scrofimicrobium. Our data suggest that Proteus, Scrofimicrobium, Corynebacterium, Vagococcus_B, Lysinibacillus_304693 (all LD), and Paenibacillus_J_366884 (WT) are lineage-specific rather than members of a common core (> 90% prevalence in either LD or WT, with prevalence significantly different between lines (p ≤ 0.05)). Positive correlations were observed between several core genera and larval performance in LD, typical of a highly optimized lab-adapted line. Interestingly, only members of the genus Providencia appeared to play a crucial role in most aspects of larval performance in both genetic lineages. Conclusion: Our study demonstrates that the gut microbiota of BSFL is influenced by genetic factors, diet composition, larval age, and their interactions. We identified a distinct lineage-specific core microbiota, emphasizing genetic background's role. Future studies should apply a standardized high prevalence threshold of at least > 90% unless there is a valid reason for relaxation or sample exclusion. The consistent association of Providencia spp. with larval performance across both genetic lines highlights their crucial role in the BSFL gut ecosystem. [ABSTRACT FROM AUTHOR]

Objective: Livestock droppings cause some environmental problems, but they have the potential to be used as effective biomass resources. The black soldier fly (BSF), Hermetia illucens (Diptera: Stratiomyidae), is suitable for efficiently processing such resources. By using BSF larvae for the disposal of livestock droppings, we can obtain two valuable products: protein resources and organic fertilizer. However, there is insufficient research on the digestive enzymes suitable for processing this waste. Here, we aimed to construct an efficient BSF processing system using livestock droppings, and we explored the digestive enzymes involved in this process. Results: First, we investigated the characteristics of transcripts expressed in the midgut of BSF larvae and found that immune response-related genes were expressed in the midgut. Then, we investigated digestive enzymes and identified a novel serine protease, HiBrachyurin, whose mRNA was highly expressed in the posterior midgut when BSF larvae fed on horse droppings. Despite the low protein content of horse droppings, larvae that fed on horse droppings accumulated more protein than those in the other groups. Therefore, HiBrachyurin may contribute to digestibility in the early stage of protein degradation in BSF larvae fed on horse droppings. [ABSTRACT FROM AUTHOR]

Background: This study aimed to evaluate the effects of Hermetia illucens (Black soldier fly-BSF) and Tenebrio molitor (Yellow mealworm-YMW) live larvae as a new nutritional model on duck's gut health, considering gut histomorphometry, mucin composition, cytokines transcription levels, and microbiota. A total of 126, 3-days-old, females Muscovy ducks were randomly allotted to three dietary treatments (6 replicates/treatment, 7 birds/pen): (i) C: basal diet; (ii) BSF: C + BSF live larvae; (iii) YMW: C + YMW live larvae. BSF and YMW live larvae were administered on top of the basal diet, based on the 5% of the expected daily feed intake. The live weight, average daily gain, average daily feed intake and feed conversion ratio were evaluated for the whole experimental period. On day 52, 12 ducks/treatment (2 birds/replicate) were slaughtered and samples of duodenum, jejunum, ileum, spleen, liver, thymus and bursa of Fabricius were collected for histomorphometry. Mucin composition was evaluated in the small intestine through histochemical staining while jejunal MUC-2 and cytokines transcription levels were evaluated by rt-qPCR. Cecal microbiota was also analyzed by means of 16 S rRNA gene sequencing. Results: Birds' growth performance and histomorphometry were not influenced by diet, with a proximo-distal decreasing gradient from duodenum to ileum (p < 0.001), respecting the physiological gut development. Mucin staining intensity and MUC-2 gene expression did not vary among dietary treatments, even though mucin intensity increased from duodenum to ileum, according to normal gut mucus physiology (p < 0.001). Regarding local immune response, IL-6 was higher in YMW group when compared to the other groups (p = 0.009). Insect live larvae did not affect cecal microbiota diversity, but BSF and YMW groups showed a higher presence of Helicobacter, Elusimicrobium, and Succinatimonas and a lower abundance of Coriobacteriaceae and Phascolarctobacterium compared to C birds (p < 0.05). Conclusions: The use of BSF and YMW live larvae as new nutritional model did not impair gut development and mucin composition of Muscovy ducks, but slightly improved the intestinal immune status and the microbiota composition by enhancing regulatory cytokine IL-6 and by increasing minor Operational Taxonomic Units (OTUs) involved in short-chain fatty acids production. [ABSTRACT FROM AUTHOR]

Background: The black soldier fly (BSF, Hermetia illucens L.) is one of the most promising insects for bioconversion of organic waste, which often carry a high microbial load with potential foodborne pathogens. Although horizontal transmission (from rearing substrate to larvae) has been extensively studied, less is known about vertical transmission of microorganisms, and particularly of foodborne pathogens, across different BSF life stages. Results: This study investigated the microbial dynamics and vertical transmission of Escherichia coli across different life stages (larvae, prepupae, pupae and adults) of one BSF life cycle and its associated substrate (chicken feed) and frass, based on a combination of general microbial counts (based on culture-dependent techniques) and the bacterial community composition (based on 16S rRNA gene sequencing). Multiple interactions between the microbiota of the substrate, frass and BSF larvae were affirmed. The larvae showed relative consistency among both the microbial counts and bacterial community composition. Diversification of the bacterial communities started during the pupal stage, while most notable changes of the microbial counts and bacterial community compositions occurred during metamorphosis to adults. Furthermore, vertical transmission of E. coli was investigated after substrate inoculation with approximately 7.0 log cfu/g of kanamycin-resistant E. coli, and monitoring E. coli counts from larval to adult stage. Although the frass still contained substantial levels of E. coli (> 4.5 log cfu/g) and E. coli was taken up by the larvae, limited vertical transmission of E. coli was observed with a decreasing trend until the prepupal stage. E. coli counts were below the detection limit (1.0 log cfu/g) for all BSF samples from the end of the pupal stage and the adult stage. Additionally, substrate inoculation of E. coli did not have a substantial impact on the bacterial community composition of the substrate, frass or different BSF life stages. Conclusions: The fluctuating microbial counts and bacterial community composition underscored the dynamic character of the microbiota of BSF life stages. Additionally, vertical transmission throughout one BSF life cycle was not observed for E. coli. Hence, these findings paved the way for future case studies on vertical transmission of foodborne pathogens across consecutive BSF life stages or other insect species. [ABSTRACT FROM AUTHOR]

Background: The world faces a major infectious disease challenge. Interest in the discovery, design, or development of antimicrobial peptides (AMPs) as an alternative approach for the treatment of bacterial infections has increased. Insects are a good source of AMPs which are the main effector molecules of their innate immune system. Black Soldier Fly Larvae (BSFL) are being developed for large-scale rearing for food sustainability, waste reduction and as sustainable animal and fish feed. Bioinformatic studies have suggested that BSFL have the largest number of AMPs identified in insects. However, most AMPs identified in BSF have not yet undergone antimicrobial evaluation but are promising leads to treat critical infections. Results: Jg7197.t1, Jg7902.t1 and Jg7904.t1 were expressed into the haemolymph of larvae following infection with Salmonella enterica serovar Typhimurium and were predicted to be AMPs using the computational tool ampir. The genes encoding these proteins were within 2 distinct clusters in chromosome 1 of the BSF genome. Following removal of signal peptides, predicted structures of the mature proteins were superimposed, highlighting a high degree of structural conservation. The 3 AMPs share primary sequences with proteins that contain a Kunitz-binding domain; characterised for inhibitory action against proteases, and antimicrobial activities. An in vitro antimicrobial screen indicated that heterologously expressed SUMO-Jg7197.t1 and SUMO-Jg7902.t1 did not show activity against 12 bacterial strains. While recombinant SUMO-Jg7904.t1 had antimicrobial activity against a range of Gram-negative and Gram-positive bacteria, including the serious pathogen Pseudomonas aeruginosa. Conclusions: We have cloned and purified putative AMPs from BSFL and performed initial in vitro experiments to evaluate their antimicrobial activity. In doing so, we have identified a putative novel defensin-like AMP, Jg7904.t1, encoded in a paralogous gene cluster, with antimicrobial activity against P. aeruginosa. [ABSTRACT FROM AUTHOR]

Background: We had earlier described the growth-promoting and -depressive effects of replacing soybean meal (SBM) with low (12.5% and 25%) and high (50% and 100%) inclusion levels of black soldier fly larvae meal (BSFLM), respectively, in Ross x Ross 708 broiler chicken diets. Herein, using 16S rRNA gene amplicon sequencing, we investigated the effects of replacing SBM with increasing inclusion levels (0-100%) of BSFLM in broiler diets on the cecal bacterial community composition at each growth phase compared to broilers fed a basal corn-SBM diet with or without the in-feed antibiotic, bacitracin methylene disalicylate (BMD). We also evaluated the impact of low (12.5% and 25%) inclusion levels of BSFLM (LIL-BSFLM) on the prevalence of selected antimicrobial resistance genes (ARGs) in litter and cecal samples from 35-day-old birds. Results: Compared to a conventional SBM-based broiler chicken diet, high (50 to100%) inclusion levels of BSFLM (HIL-BSFLM) significantly altered the cecal bacterial composition and structure, whereas LIL-BSFLM had a minimal effect. Differential abundance analysis further revealed that the ceca of birds fed 100% BSFLM consistently harbored a ~ 3 log-fold higher abundance of Romboutsia and a ~ 2 log-fold lower abundance of Shuttleworthia relative to those fed a BMD-supplemented control diet at all growth phases. Transient changes in the abundance of several potentially significant bacterial genera, primarily belonging to the class Clostridia, were also observed for birds fed HIL-BSFLM. At the finisher phase, Enterococci bacteria were enriched in the ceca of chickens raised without antibiotic, regardless of the level of dietary BSFLM. Additionally, bacitracin (bcrR) and macrolide (ermB) resistance genes were found to be less abundant in the ceca of chickens fed antibiotic-free diets, including either a corn-SBM or LIL-BSFLM diet. Conclusions: Chickens fed a HIL-BSFLM presented with an imbalanced gut bacterial microbiota profile, which may be linked to the previously reported growth-depressing effects of a BSFLM diet. In contrast, LIL-BSFLM had a minimal effect on the composition of the cecal bacterial microbiota and did not enrich for selected ARGs. Thus, substitution of SBM with low levels of BSFLM in broiler diets could be a promising alternative to the antibiotic growth promoter, BMD, with the added-value of not enriching for bacitracin- and macrolide-associated ARGs. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: There is a global need to develop new therapies to treat infectious diseases and tackle the rise in antimicrobial resistance. To date, the larvae of the Black Solider Fly, Hermetia illucens, have the largest repertoire of antimicrobial peptides derived from insects. Antimicrobial peptides are of particular interest in the exploration of alternative antimicrobials due to their potent action and reduced propensity to induce resistance compared with more traditional antibiotics. Results: The predicted attacin from H. illucens, Hill_BB_C10074, was first identified in the transcriptome of H. illucens populations that had been fed a plant-oil based diet. In this study, recombinant Hill_BB_C10074 (500 µg/mL), was found to possess potent antimicrobial activity against the serious Gram-negative pathogen, Pseudomonas aeruginosa. Sequence and structural homology modelling predicted that Hill_BB_C10074 formed a homotrimeric complex that may form pores in the Gram-negative bacterial outer membrane. In vitro experiments defined the antimicrobial action of Hill_BB_C10074 against P. aeruginosa and transmission electron microscopy and electrochemical impedance spectroscopy confirmed the outer membrane disruptive power of Hill_BB_C10074 which was greater than the clinically relevant antibiotic, polymyxin B. Conclusions: Combining predictive tools with in vitro approaches, we have characterised Hill_BB_C10074 as an important insect antimicrobial peptide and promising candidate for the future development of clinical antimicrobials. [ABSTRACT FROM AUTHOR]

Background: In the last few years, considerable attention has been focused on the plastic-degrading capability of insects and their gut microbiota in order to develop novel, effective, and green strategies for plastic waste management. Although many analyses based on 16S rRNA gene sequencing are available, an in-depth analysis of the insect gut microbiome to identify genes with plastic-degrading potential is still lacking. Results: In the present work, we aim to fill this gap using Black Soldier Fly (BSF) as insect model. BSF larvae have proven capability to efficiently bioconvert a wide variety of organic wastes but, surprisingly, have never been considered for plastic degradation. BSF larvae were reared on two widely used plastic polymers and shotgun metagenomics was exploited to evaluate if and how plastic-containing diets affect composition and functions of the gut microbial community. The high-definition picture of the BSF gut microbiome gave access for the first time to the genomes of culturable and unculturable microorganisms in the gut of insects reared on plastics and revealed that (i) plastics significantly shaped bacterial composition at species and strain level, and (ii) functions that trigger the degradation of the polymer chains, i.e., DyP-type peroxidases, multicopper oxidases, and alkane monooxygenases, were highly enriched in the metagenomes upon exposure to plastics, consistently with the evidences obtained by scanning electron microscopy and 1H nuclear magnetic resonance analyses on plastics. Conclusions: In addition to highlighting that the astonishing plasticity of the microbiota composition of BSF larvae is associated with functional shifts in the insect microbiome, the present work sets the stage for exploiting BSF larvae as "bioincubators" to isolate microbial strains and enzymes for the development of innovative plastic biodegradation strategies. However, most importantly, the larvae constitute a source of enzymes to be evolved and valorized by pioneering synthetic biology approaches. 5errh-AhYWWkX5YWmGozvJ Video Abstract [ABSTRACT FROM AUTHOR]

Bioconversion using insects is a promising strategy to convert organic waste (catering leftovers, harvest waste, food processing byproducts, etc.) into biomass that can be used for multiple applications, turned into high added-value products, and address environmental, societal and economic concerns. Due to its ability to feed on a tremendous variety of organic wastes, the black soldier fly (Hermetia illucens) has recently emerged as a promising insect for bioconversion of organic wastes on an industrial scale. A growing number of studies have highlighted the pivotal role of the gut microbiota in the performance and health of this insect species. This review aims to provide a critical overview of current knowledge regarding the functional diversity of the gut microbiota of H. illucens, highlighting its importance for bioconversion, food safety and the development of new biotechnological tools. After providing an overview of the different strategies that have been used to outline the microbial communities of H. illucens, we discuss the diversity of these gut microbes and the beneficial services they can provide to their insect host. Emphasis is placed on technical strategies and aspects of host biology that require special attention in the near future of research. We also argue that the singular digestive capabilities and complex gut microbiota of H. illucens make this insect species a valuable model for addressing fundamental questions regarding the interactions that insects have evolved with microorganisms. By proposing new avenues of research, this review aims to stimulate research on the microbiota of a promising insect to address the challenges of bioconversion, but also fundamental questions regarding bacterial symbiosis in insects. [ABSTRACT FROM AUTHOR]

Background: In contrast to protein-rich insect meal, the feed potential of insect fat is generally less explored and knowledge about the suitability of insect fat as a fat source specifically in broiler diets is still limited. In view of this, the present study aimed to comprehensively investigate the effect of partial (50%) and complete replacement of soybean oil with insect fat from Hermetia illucens (HI) larvae in broiler diets on performance, fat digestibility, cecal microbiome, liver transcriptome and liver and plasma lipidomes. Thus, 100 male, 1-day-old Cobb 500 broilers were randomly assigned to three groups and fed three different diets with either 0 (group HI-0, n = 30), 2.5% (group HI-2.5, n = 35) or 5.0% (HI-5.0, n = 35) Hermetia illucens (HI) larvae fat for 35 d. Results: Body weight gain, final body weight, feed intake, and feed:gain ratio during the whole period and apparent ileal digestibility coefficient for ether extract were not different between groups. Cecal microbial diversity did not differ between groups and taxonomic analysis revealed differences in the abundance of only four low-abundance bacterial taxa among groups; the abundances of phylum Actinobacteriota, class Coriobacteriia, order Coriobacteriales and family Eggerthellaceae were lower in group HI-5.0 compared to group HI-2.5 (P < 0.05). Concentrations of total and individual short-chain fatty acids in the cecal digesta were not different between the three groups. Liver transcriptomics revealed a total of 55 and 25 transcripts to be differentially expressed between groups HI-5.0 vs. HI-0 and groups HI-2.5 vs. HI-0, respectively (P < 0.05). The concentrations of most lipid classes, with the exception of phosphatidylethanolamine, phosphatidylglycerol and lysophosphatidylcholine in the liver and cholesterylester and ceramide in plasma (P < 0.05), and of the sum of all lipid classes were not different between groups. Conclusions: Partial and complete replacement of soybean oil with HI larvae fat in broiler diets had no effect on growth performance and only modest, but no adverse effects on the cecal microbiome and the metabolic health of broilers. This suggests that HI larvae fat can be used as an alternative fat source in broiler diets, thereby, making broiler production more sustainable. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Increasing demand for high-value fish species and pressure on forage fish is challenging aquaculture to ensure sustainable growth by replacing protein sources in aquafeeds with plant and terrestrial animal proteins, without compromising the economic value and quality of the final fish product. In the present study, the effects of a plant protein-based diet (CV), two plant-based diets in which graded amounts of plan protein mixtures were replaced with Hermetia illucens meal alone (VH10) or in combination with poultry by-product meal (PBM) (VH10P30), a fishmeal (FM) diet (CF) and an FM diet supplemented with H. illucens (FH10) on growth performance, gut health and homeostasis of farmed subadult European seabass were tested and compared. Results: Fish fed the VH10 and VH10P30 diets showed the highest specific growth rates and lowest feed conversion ratios among the tested groups. Expectedly, the best preservation of PI morphology was observed in fish fed the CF or FH10 diets, while fish fed the CV diet exhibited significant degenerative changes in the proximal and distal intestines. However, PBM supplementation mitigated these effects and significantly improved all gut morphometric parameters in the VH10P30 group. Partial substitution of the plant mixture with insect meal alone or PBM also induced most BBM genes and activated BBM enzymes, suggesting a beneficial effect on intestinal digestive/absorption functions. Regarding intestinal microbiota, fish fed diets containing H. illucens meal (FH10, VH10, VH10P30) had the highest richness of bacterial communities and abundance of beneficial genera such as Lactobacillus and Bacillus. On the other hand, fish fed CV had the highest microbial diversity but lost a significant component of fish intestinal microbiota, the phylum Bacteroidetes. Finally, skin pigmentation most similar to that of farmed or even wild seabass was also observed in the fish groups fed CF, FH10 or VH10P30. Conclusion: Plant-based diets supplemented with PBM and H. illucens pupae meal have great potential as alternative diets for European seabass, without affecting growth performance, gut homeostasis, or overall fitness. This also highlights the importance of animal proteins in diets of European seabass, as the addition of a small amount of these alternative animal protein sources significantly improved all measured parameters. [ABSTRACT FROM AUTHOR]

Background: The possibility of partially replacing soybean meal (SBM) with Hermetia illucens (HI) defatted larvae meal in broiler nutrition has frequently been suggested. For sustainability reasons, however, the larvae fat produced during defatting should also be used and could be particularly beneficial regarding gut health due to its fatty acid composition. To evaluate the suitability of HI larvae as protein and fat source, a 2 × 3 factorial arrangement with two types of protein, i.e. SBM (S) or SBM and 15% of its crude protein replaced by HI larvae meal (L), and three levels of fat sources, namely 0 (0 L), 50% (50 L) or 100% HI larvae fat (100 L) at the expense of soybean oil was applied. Results: In the starter phase, an interaction showed higher body weight (BW), average daily gain (ADG) and improved feed conversion ratio (FCR) if 50% or 100% HI larvae fat was fed with HI larvae meal. Moreover, BW, ADG and FCR improved when feeding HI larvae meal as protein source. Additionally, we observed an increased average daily feed intake in the grower, finisher, and overall phase in the L groups and an improved FCR in 0 L compared to 50 L groups during the overall period. Regarding apparent ileal digestibility, HI larvae meal feeding increased dry matter, organic matter, and fat digestibility. Feeding HI larvae meal as protein source decreased the concentrations of agmatine, spermidine, spermine and ammonia in the caecal digesta, whereas fat source affected agmatine with higher concentrations in 50 L compared to 0 L in the colonic digesta. In contrast, caecal ethanolamine concentrations increased in HI larvae meal groups compared to SBM. Caecal butyric acid concentrations decreased with HI larvae meal feeding. An interaction was found for the jejunal villus area, being higher in L + 100 L compared to S + 100 L. Furthermore, L groups had greater villus width. Conclusions: A partial replacement of SBM with HI larvae meal and soybean oil with HI larvae fat in broiler diets without impairing animal performance or gut health seems possible. Feeding HI larvae meal affected broiler performance positively in the starter phase and improved apparent ileal digestibility. [ABSTRACT FROM AUTHOR]

In recent years, interest in the larvae of black soldier fly (BSF) (Hermetia illucens) as a sustainable protein resource for livestock feed has increased considerably. However, knowledge on the nutritional and physiological aspects of this insect, especially compared to other conventional farmed animals is scarce. This review presents a critical comparison of data on the growth potential and efficiency of the BSF larvae (BSFL) compared to conventional monogastric livestock species. Advantages of BSFL over other monogastric livestock species includes their high growth rate and their ability to convert low-grade organic waste into high-quality protein and fat-rich biomass suitable for use in animal feed. Calculations using literature data suggest that BSFL are more efficient than broilers, pigs and fish in terms of conversion of substrate protein into body mass, but less efficient than broilers and fish in utilization of substrate gross energy to gain body mass. BSFL growth efficiency varies greatly depending on the nutrient quality of their dietary substrates. This might be associated with the function of their gastrointestinal tract, including the activity of digestive enzymes, the substrate particle characteristics, and their intestinal microbial community. The conceived advantage of BSFL having an environmental footprint better than conventional livestock is only true if BSFL is produced on low-grade organic waste and its protein would directly be used for human consumption. Therefore, their potential role as a new species to better close nutrient cycles in agro-ecological systems needs to be reconsidered, and we conclude that BSFL is a complementary livestock species efficiently utilizing organic waste that cannot be utilized by other livestock. In addition, we provide comparative insight into morpho-functional aspects of the gut, characterization of digestive enzymes, gut microbiota and fiber digestion. Finally, current knowledge on the nutritional utilization and requirements of BSFL in terms of macro- and micro-nutrients is reviewed and found to be rather limited. In addition, the research methods to determine nutritional requirements of conventional livestock are not applicable for BSFL. Thus, there is a great need for research on the nutrient requirements of BSFL. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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 FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: The black soldier fly (Hermetia illucens) has significant economic potential. The larvae can be used in financially viable waste management systems, as they are voracious feeders able to efficiently convert low-quality waste into valuable biomass. However, most studies on H. illucens in recent decades have focused on optimizing their breeding and bioconversion conditions, while information on their biology is limited. Methods: About 200 fifth instar well-fed larvae were sacrificed in this work. The liquid chromatography-tandem mass spectrometry and scanning electron microscopy were employed in this study to perform a proteomic and ultrastructural analysis of the peritrophic matrix (PM) of H. illucens larvae. Results: A total of 565 proteins were identified in the PM samples of H. illucen, of which 177 proteins were predicted to contain signal peptides, bioinformatics analysis and manual curation determined 88 proteins may be associated with the PM, with functions in digestion, immunity, PM modulation, and others. The ultrastructure of the H. illucens larval PM observed by scanning electron microscopy shows a unique diamond-shaped chitin grid texture. Conclusions: It is the first and most comprehensive proteomics research about the PM of H. illucens larvae to date. All the proteins identified in this work has been discussed in details, except several unnamed or uncharacterized proteins, which should not be ignored and need further study. A comparison of the ultrastructure between H. illucens larval PM and those of other insects as observed by SEM indicates that the PM displays diverse textures on an ultra-micro scale and we suscept a unique diamond-shaped chitin grid texture may help H. illucens larval to hold more food. This work deepens our understanding of the molecular architecture and ultrastructure of the H. illucens larval PM. [ABSTRACT FROM AUTHOR]

Background: Intestinal digesta is commonly used for studying responses of microbiota to dietary shifts, yet evidence is accumulating that it represents an incomplete view of the intestinal microbiota. The present work aims to investigate the differences between digesta- and mucosa-associated intestinal microbiota in Atlantic salmon (Salmo salar) and how they may respond differently to dietary perturbations. In a 16-week seawater feeding trial, Atlantic salmon were fed either a commercially-relevant reference diet or an insect meal diet containing ~ 15% black soldier fly (Hermetia illucens) larvae meal. The digesta- and mucosa-associated distal intestinal microbiota were profiled by 16S rRNA gene sequencing. Results: Regardless of diet, we observed substantial differences between digesta- and mucosa-associated intestinal microbiota. Microbial richness and diversity were much higher in the digesta than the mucosa. The insect meal diet altered the distal intestinal microbiota resulting in higher microbial richness and diversity. The diet effect, however, depended on the sample origin. Digesta-associated intestinal microbiota showed more pronounced changes than the mucosa-associated microbiota. Multivariate association analyses identified two mucosa-enriched taxa, Brevinema andersonii and Spirochaetaceae, associated with the expression of genes related to immune responses and barrier function in the distal intestine, respectively. Conclusions: Our data show that salmon intestinal digesta and mucosa harbor microbial communities with clear differences. While feeding insects increased microbial richness and diversity in both digesta- and mucosa-associated intestinal microbiota, mucosa-associated intestinal microbiota seems more resilient to variations in the diet composition. To fully unveil the response of intestinal microbiota to dietary changes, concurrent profiling of digesta- and mucosa-associated intestinal microbiota is recommended whenever feasible. Specific taxa enriched in the intestinal mucosa are associated to gene expression related to immune responses and barrier function. Detailed studies are needed on the ecological and functional significance of taxa associated to intestinal microbiota dwelling on the mucosa. [ABSTRACT FROM AUTHOR]

Background: The black soldier fly (Diptera: Stratiomyidae, Hermetia illucens) is renowned for its bioconversion ability of organic matter, and is the worldwide most widely used source of insect protein. Despite varying extensively in morphology, it is widely assumed that all black soldier flies belong to the same species, Hermetia illucens. We here screened about 600 field-collected and cultured flies from 39 countries and six biogeographic regions to test this assumption based on data for three genes (mitochondrial COI, nuclear ITS2 & 28S rDNA) and in order to gain insights into the phylogeography of the species. Results: Our study reveals a surprisingly high level of intraspecific genetic diversity for the mitochondrial barcoding gene COI (divergences up to 4.9%). This level of variability is often associated with the presence of multiple species, but tested nuclear markers (ITS2 and 28S rDNA) were invariant and fly strain hybridization experiments under laboratory conditions revealed reproductive compatibility. COI haplotype diversity is not only very high in all biogeographic regions (56 distinct haplotypes in total), but also in breeding facilities and research centers from six continents (10 haplotypes: divergences up to 4.3%). The high genetic diversity in fly-breeding facilities is mostly likely due to many independent acquisitions of cultures via sharing and/or establishing new colonies from field-collected flies. However, explaining some of the observed diversity in several biogeographic regions is difficult given that the origin of the species is considered to be New World (32 distinct haplotypes) and one would expect severely reduced genetic diversity in the putatively non-native populations in the remaining biogeographic regions. However, distinct, private haplotypes are known from the Australasian (N = 1), Oriental (N = 4), and the Eastern Palearctic (N = 4) populations. We reviewed museum specimen records and conclude that the evidence for introductions is strong for the Western Palearctic and Afrotropical regions which lack distinct, private haplotypes. Conclusions: Based on the results of this paper, we urge the black soldier fly community to apply molecular characterization (genotyping) of the fly strains used in artificial fly-breeding and share these data in research publications as well as when sharing cultures. In addition, fast-evolving nuclear markers should be used to reconstruct the recent invasion history of the species. [ABSTRACT FROM AUTHOR]

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 FROM AUTHOR]