A novel family of defensin-like peptides from Hermetia illucens with antibacterial properties.

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.

Recombinant expression and characterization of the endochitinase Chit36-TA from Trichoderma asperellum in Komagataella phaffii for chitin degradation of black soldier fly exuviae.

The natural polymer chitin is an abundant source for valuable N-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni-NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36 kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50 °C, while it maintained > 93% activity up to 60 °C. The chitinase was thermostable up to 45 °C and exhibited ~ 50% activity after a 15 min incubation at 57 °C. Chit36-TA had a maximum specific enzyme activity of 50 nkat/mg with a Km value of 289 µM with 4-methylumbelliferyl-N,N',N″-triacetyl-ß-chitotrioside as substrate. Most tested cations, organic solvents and reagents were well-tolerated by the endochitinase, except for SDS (1 mM), Cu2+ (10 mM) and Mn2+ (10 mM), which had stronger inhibitory effects with residual activities of 3, 41 and 28%, respectively. With a degree of hydrolysis of 32% applying colloidal shrimp chitin (1% (w/v)) and 12% on insect larvae (1% (w/v)) after 24 h, the endochitinase was found to be suitable for the conversion of colloidal chitin as well as chitin from black soldier fly larvae into water-soluble N-acetylchitooligosaccharides. To prove scalability, a bioreactor process was developed in which a 55-fold higher enzyme activity of 49 µkat/l and a tenfold higher protein expression of 1258 mg/l were achieved.

Synergistic impacts of antibiotics and heavy metals on Hermetia illucens: Unveiling dynamics in larval gut bacterial communities and microbial metabolites.

Hermetia illucens larvae showcases remarkable bioremediation capabilities for both antibiotics and heavy metal contaminants. However, the distinctions in larval intestinal microbiota arising from the single and combined effects of antibiotics and heavy metals remain poorly elucidated. In this study, we delved into the details of larval intestinal bacterial communities and microbial metabolites when exposed to single and combined contaminants of oxytetracycline (OTC) and hexavalent chromium (Cr(VI)). After conversion, single contaminant-spiked substrate showed 75.5% of OTC degradation and 95.2% of Cr(VI) reductiuon, while combined contaminant-spiked substrate exhibited 71.3% of OTC degradation and 93.4% of Cr(VI) reductiuon. Single and combined effects led to differences in intestinal bacterial communities, mainly reflected in the genera of Enterococcus, Pseudogracilibacillus, Gracilibacillus, Wohlfahrtiimonas, Sporosarcina, Lysinibacillus, and Myroide. Moreover, these effects also induced differences across various categories of microbial metabolites, which categorized into amino acid and its metabolites, benzene and substituted derivatives, carbohydrates and its metabolites, heterocyclic compounds, hormones and hormone-related compounds, nucleotide and its metabolites, and organic acid and its derivatives. In particular, the differences induced OTC was greater than that of Cr(VI), and combined effects increased the complexity of microbial metabolism compared to that of single contaminant. Correlation analysis indicated that the bacterial genera, Preudogracilibacillus, Enterococcus, Sporosarcina, Lysinibacillus, Wohlfahrtiimonas, Ignatzschineria, and Fusobacterium exhibited significant correlation with significant differential metabolites, these might be used as indicators for the resistance and bioremediation of OTC and Cr(VI) contaminants. These findings are conducive to further understanding that the metabolism of intestinal microbiota determines the resistance of Hermetia illucens to antibiotics and heavy metals.

Treatment of landfill leachate by black soldier fly (Hermetia illucens L.) larvae and the changes of intestinal microbial community.

Black soldier fly larvae (BSFL) (Hermetia illucens) are commonly used to treat organic waste. This work aims to evaluate the transformation effect, heavy metal migration, and alterations in the gut microbiota of BSFL in addition to treating landfill leachate (LL) with BSFL. We found that BSFL may grow in various landfill leachate concentrations without obvious toxicity and growth inhibition. In addition, the results indicated a significant increase in the content of ammonia nitrogen and the activity of urease and ß-glucosidase (ß-GC) in LL, increased from 2570.17 mg/L to 5853.67 mg/L, 1859.17 mg/(g·d) to 517,177.98 mg/(g·d), 313.73 µg/(g·h) to 441.91 µg/(g·h) respectively. Conversely, the content of total nitrogen (TN) and total organic carbon (TOC) decreased in LL, decreasing by 31.24% and 29.45% respectively. Heavy metals are accumulated in the leachate by the BSFL to differing degrees, the descending sequence of accumulation is Cd > As > Cu > Cr. As dropped by 26.0%, Cd increased by 22.6%, Cu reduced by 5.23%, and Cr increased by 317.1% in the remaining matrix. The concentration of heavy metals satisfies the organic fertilizers' limit index (NY/T1978). The diversity of intestinal microorganisms in BSFL decreased, from 2819 OTUs to 2338 OTUs, with Providencia and Morganella emerging as the core flora. The gene abundance of nitrogen metabolism in the microbiota increased significantly. The TOC, ß-GC, and Copper (Cu) content in BSFL correlated significantly with the gut microbiota. In Summary, this study revealed the treatment effect of BSFL on LL, the migration of heavy metals, and changes in the intestinal microorganisms of BSFL. The content of heavy metals in BSFL was found to be much lower than the upper limit of feed protein raw materials, demonstrating that BSFL is a sustainable method to treat LL.

Near-infrared spectroscopy (NIRS) for monitoring the nutritional composition of black soldier fly larvae (BSFL) and frass.

BACKGROUND: The demand for protein obtained from animal sources is growing rapidly, as is the necessity for sustainable animal feeds. The use of black soldier fly larvae (BSFL) reared on organic side streams as sustainable animal feed has been receiving attention lately. This study assessed the ability of near-infrared spectroscopy (NIRS) combined with chemometrics to evaluate the nutritional profile of BSFL instars (fifth and sixth) and frass obtained from two different diets, namely soy waste and customised bread-vegetable diet. Partial least squares (PLS) regression with leave one out cross-validation was used to develop models between the NIR spectral data and the reference analytical methods. RESULTS: Calibration models with good [coefficient of determination in calibration (Rcal 2 ): 0.90; ratio of performance to deviation (RPD) value: 3.6] and moderate (Rcal 2 : 0.76; RPD value: 2.1) prediction accuracy was observed for acid detergent fibre (ADF) and total carbon (TC), respectively. However, calibration models with moderate accuracy were observed for the prediction of crude protein (CP) (Rcal 2 : 0.63; RPD value: 1.4), crude fat (CF) (Rcal 2 : 0.70; RPD value: 1.6), neutral detergent fibre (NDF) (Rcal 2 : 0.60; RPD value: 1.6), starch (Rcal 2 : 0.52; RPD value: 1.4), and sugars (Rcal 2 : 0.52; RPD value: 1.4) owing to the narrow or uneven distribution of data over the range evaluated. CONCLUSION: The near-infrared (NIR) calibration models showed a good to moderate prediction accuracy for the prediction of ADF and TC content for two different BSFL instars and frass reared on two different diets. However, calibration models developed for predicting CP, CF, starch, sugars and NDF resulted in models with limited prediction accuracy. © 2023 Society of Chemical Industry.

Selective cytotoxicity of standardised n-hexane extract of black soldier flies' larvae on cancerous skin cells mitochondria isolated from rat model of melanoma.

INTRODUCTION: Melanoma is known as a highly lethal cancer. In melanoma cells, apoptosis signalling which relies heavily on the acute activity of mitochondria and reactive oxygen species (ROS) formation is suppressed. Our previous studies on natural compounds on melanoma suggested that mitochondria are a potential target for the melanoma treatment by selective cytotoxic effect of them. The black soldier fly is an important environmental protectant insect that based on recent studies induces apoptosis in liver and colorectal carcinoma cells through the activation of caspase 3, 8, and 9 and ultimately inhibits the growth of cancer cells. PURPOSE: This study was designed to evaluate the selective apoptotic effect of the n-hexane BSFL extract (BSFLE) on skin mitochondria. MATERIALS AND METHODS: The mitochondria isolated from melanoma cells were treated with various concentrations (1500, 3000, and 6000 µg/ml) of n-hexane BSFLE Then MTT viability assay, ROS determination, Mitochondrial Membrane Potential (MMP), mitochondrial swelling, cytochrome c release determination, and % apoptosis were performed. RESULTS: MTT assay showed that different concentrations of n-hexane BSFLE significantly (P < 0.05) decreased the SDH activity in cancerous skin mitochondria with the IC50. The ROS production and mitochondrial swelling results also showed that all concentrations of BSFL extracts significantly increased. MMP decline and the release of cytochrome c in cancer groups mitochondria. BSFLE increased apoptosis on melanoma cells. DISCUSSION AND CONCLUSION: It is suggested that n-hexane BSFLE compounds selectively induce a cascade of proapoptotic events that are probably defective in cancer cells. Most of these compounds target the mitochondrial transient pore caused by disruption of the mitochondrial respiratory chain. These events lead to disruption of the temporary permeability of mitochondria, swelling of mitochondria and finally the formation of apoptosome in the cytosol.

Increasing levels of fishmeal replacement by defatted black soldier fly larvae meal reduced growth performance without affecting fillet quality in largemouth bass (Micropterus salmoides).

A 90-day feeding trial was conducted to evaluate the effects of replacing dietary fishmeal (FM) with defatted black soldier fly larvae meal (BSFL) on the growth performance and fillet quality of largemouth bass (Micropterus salmoides). The largemouth bass was divided into six groups (BSFL0, BSFL15, BSFL30, BSFL45, BSFL75, and BSFL100) and fed six isonitrogenous(CP 50%, 508 g/kg) and isolipid (CL 9%, 124 g/kg) diets, in which 0, 15%, 30%, 45%, 75%, and 100% of the fishmeal was replaced with BSFL, respectively. The results showed that the final body weight (FBW) and specific growth rate (SGR) of the largemouth bass decreased with increasing BSFL content, and they were significantly lower in BSFL75 than in BSFL0. The weight gain rate (WGR) decreased with increasing BSFL content and the feed conversion ratio (FCR) of largemouth bass increased with increasing BSFL content. The saturated fatty acids (SFAs) and n-3 polyunsaturated fatty acids (PUFA) contents of the largemouth bass fillet significantly decreased, and the n-6 PUFA content of the largemouth bass fillets significantly increased with increasing dietary BSFL. The fillet b* significantly decreased with increasing BSFL content. The biological parameters, fillet proximate nutrient composition, fillet amino acid composition, skin color, and fillet texture of the largemouth bass were not affected by the replacement of BSFL. Transcriptomic analysis revealed that BSFL replacement of FM affects the immune system and metabolic processes of largemouth bass through signaling pathways such as complement and coagulation cascades, the PPAR signaling pathway, cholesterol metabolism, and fat digestion and absorption. In conclusion, a replacement level lower than 45% BSFL was suggested for the overall growth and fillet quality of largemouth bass.

Near Infrared Spectroscopy for Prediction of Yeast and Mould Counts in Black Soldier Fly Larvae, Feed and Frass: A Proof of Concept.

The use of black soldier fly larvae (BSFL) grown on different organic waste streams as a source of feed ingredient is becoming very popular in several regions across the globe. However, information about the easy-to-use methods to monitor the safety of BSFL is a major step limiting the commercialization of this source of protein. This study investigated the ability of near infrared (NIR) spectroscopy combined with chemometrics to predict yeast and mould counts (YMC) in the feed, larvae, and the residual frass. Partial least squares (PLS) regression was employed to predict the YMC in the feed, frass, and BSFL samples analyzed using NIR spectroscopy. The coefficient of determination in cross validation (R2CV) and the standard error in cross validation (SECV) obtained for the prediction of YMC for feed were (R2cv: 0.98 and SECV: 0.20), frass (R2cv: 0.81 and SECV: 0.90), larvae (R2cv: 0.91 and SECV: 0.27), and the combined set (R2cv: 0.74 and SECV: 0.82). However, the standard error of prediction (SEP) was considered moderate (range from 0.45 to 1.03). This study suggested that NIR spectroscopy could be utilized in commercial BSFL production facilities to monitor YMC in the feed and assist in the selection of suitable processing methods and control systems for either feed or larvae quality control.

Anti-Inflammatory Activity of Black Soldier Fly Oil Associated with Modulation of TLR Signaling: A Metabolomic Approach.

Dietary intervention in the treatment of ulcerative colitis involves, among other things, modifications in fatty acid content and/or profile. For example, replacing saturated long chain fatty acids with medium chain fatty acids (MCFAs) has been reported to ameliorate inflammation. The Black Soldier Fly Larvae's (BSFL) oil is considered a sustainable dietary ingredient rich in the MCFA C12:0; however, its effect on inflammatory-related conditions has not been studied until now. Thus, the present study aimed to investigate the anti-inflammatory activity of BSFL oil in comparison to C12:0 using TLR4- or TLR2-activated THP-1 and J774A.1 cell lines and to assess its putative protective effect against dextran sulfate sodium (DSS)-induced acute colitis in mice. BSFL oil and C12:0 suppressed proinflammatory cytokines release in LPS-stimulated macrophages; however, only BSFL oil exerted anti-inflammatory activity in Pam3CSK4-stimulated macrophages. Transcriptome analysis provided insight into the possible role of BSFL oil in immunometabolism switch, involving mTOR signaling and an increase in PPAR target genes promoting fatty acid oxidation, exhibiting a discrepant mode of action compared to C12:0 treatment, which mainly affected cholesterol biosynthesis pathways. Additionally, we identified anti-inflammatory eicosanoids, oxylipins, and isoprenoids in the BSFL oil that may contribute to an orchestrated anti-inflammatory response. In vivo, a BSFL oil-enriched diet (20%) ameliorated the clinical signs of colitis, as indicated by improved body weight recovery, reduced colon shortening, reduced splenomegaly, and an earlier phase of secretory IgA response. These results indicate the novel beneficial use of BSFL oil as a modulator of inflammation.

The apparent metabolisable energy and ileal amino digestibility of black soldier fly (Hermetia illucens) larvae meal for broiler chickens.

1. Two experiments were conducted to determine the apparent metabolisable energy (AME) and standardised amino acid digestibility coefficients (SIDC) of black soldier fly larvae (BSFL) for broiler chickens.2. The BSFL contained, on a g/kg dry matter basis: crude protein, 486; crude fat, 320; ash, 58.5; neutral detergent fibre, 181; calcium, 6.8 and phosphorus, 9.1.3. In Experiment 1, an AME assay was performed wherein broilers were fed two experimental diets (a maize-soy basal diet and a test diet containing 250 g/kg BSFL) for 4 d from d 18 posthatch. The AME of BSFL was calculated based on the difference between the AME values of basal and test diets. The AME and nitrogen-corrected AME were determined to be 19.1and 18.0 MJ/kg of dry matter, respectively.4. In Experiment 2, the ileal amino acid (AA) digestibility of BSFL was determined using 22-d-old broilers by the direct method and the digesta was collected on d 25. The ratios between the AA and titanium in the diet and digesta were used to calculate the apparent digestibility and then standardised using previously published endogenous losses to estimate the SIDC of AA. The SIDC of Lys, Met, Thr, Val and average SIDC of AA in our BSFL sample were 0.85, 0.90, 0.91, 0.87 and 0.84, respectively.5. The findings showed that the BSFL meal is a good source of available energy and digestible AA, and could be a potential substitute for soybean meal in broiler diets.

Correlating black soldier fly larvae growths with soluble nutrients derived from thermally pre-treated waste activated sludge.

Black soldier fly larvae (BSFL) have been deployed to valorize various organic wastes. Nonetheless, its growth rate whilst being offered with waste activated sludge (WAS) is not promising, likely by virtue of the presence of extracellular polymeric substances' structure in WAS. In this work, the WAS were first thermally pre-treated under different treatment temperatures and durations before being administered as the feeding substrates for BSFL. The results showed the thermal pre-treatment could improve WAS palatability and subsequently, enhance the growth of BSFL especially after the pre-treatments at 75 °C and above. The highest larva weight gained was recorded at 2.16 mg/larva for the WAS sample being pre-treated at 90 °C and 16 h. Furthermore, the samples pre-treated above 75 °C also achieved higher degradation rates, indicating that the 75 °C was a threshold temperature to effectively hydrolyze the WAS. The changes of WAS characteristics, namely, (i) soluble chemical oxygen demand (SCOD), (ii) soluble carbohydrate, (iii) soluble protein, (iv) humic substances and (v) total soluble protein and humic substances, after the thermal pre-treatments were also studied in correlating with the BSFL growth. Accordingly, a model was successfully developed with the highest R2 value attained at 0.95, evidencing the SCOD was the most suitable WAS characteristic to accurately predict the BSFL growth behavior.

Enriched sewage sludge from anaerobic pre-treatment in spurring valorization potential of black soldier fly larvae.

The valorization of sewage sludge by black soldier fly larvae (BSFL) has gained attentions for sewage sludge management since the sludge can be reduced securely as well as larval biomass can be used for biorefineries application. Nevertheless, the BSFL growth was impeded while assimilating nutrition from sewage sludge due to the presence of extracellular polymeric substances (EPS) that had entrapped the essential nutrients inside. Accordingly, the pre-treatment of sewage sludge via anaerobic digestion at different pH was employed in this work to rupture the EPS structure and release more nutrients for larval growth. The results showed that larvae fed with raw sewage sludge had attained the lowest final larval weight (2.05 ± 0.38 mg/larva) as opposed to batches fed with pre-treated sewage sludges. This was because the soluble carbohydrate (more than 6.81 ± 1.31 mg of glucose/g sewage sludge) in EPS was released after anaerobic pre-treatment, facilitating larval assimilation for growth. Furthermore, it was observed that further increasing of pH for sewage sludge pre-treatment had led to lower final larval weight gained due to the inhibitory effect stemming from ammonia production at higher pH. The anaerobic pre-treatment of sewage sludge being executed at pH 3 for 8 days had achieved the highest final larval weight at 7.34 ± 0.97 mg/larva. The still low quality of sewage sludges after the pre-treatment also offered benefit, where high sewage sludge reduction and waste reduction index were recorded due to the necessity of BSFL to consume more sewage sludge in compensating the nutrients destitution in sludge. Lastly, the possibility of predicting final larval weight was successfully materialized via a statistical model derived from the multiple linear regression method. The derived model incorporated the interactive parameters of anaerobic pre-treated pH and durations at various combinations could predict the final larval weight.

The enhanced degradation behavior of oxytetracycline by black soldier fly larvae with tetracycline resistance genes in the larval gut: Kinetic process and mechanism.

Black soldier fly larvae (larvae) can digest organic wastes and degrade contaminants such as oxytetracycline (OTC). However, compared to the kinetic processes and enhanced mechanisms used in the traditional microbial degradation of OTC, those employed by larvae are largely uncharacterized. To obtain further details, a combined analysis of larval development, larval nutritional values (crude protein, crude fat and the composition of fatty acids) and the expression of tetracycline resistance genes (TRGs) in the larval gut was performed for the degradation of OTC added to substrates and for oxytetracycline bacterial residue (OBR). When the larvae were exposed to the substrates, the degradation processes were enhanced significantly (P < 0.01), with a 4.74-7.86-fold decrease in the degradation half-life (day-1) and a 3.34-5.74-fold increase in the final degradation efficiencies. This result was attributed to the abundant TRGs (with a detection rate of 35.90%∼52.14%) in the larval gut. The TRGs presented the resistance mechanisms of cellular protection and efflux pumps, which ensured that the larvae could tolerate elevated OTC concentrations. Investigation of the TRGs indicated that enzymatic inactivation enhanced OTC degradation by larvae. These findings demonstrate that the larval degradation of antibiotic contaminants is an efficient method based on abundant TRGs in the larval gut, even though OTC degradation results in OBR. In addition, a more optimized system for higher reductions in antibiotic levels and the expansion of larval bioremediation to other fields is necessary.

A sustainable and economic strategy to reduce risk antibiotic resistance genes during poultry manure bioconversion by black soldier fly Hermetia illucens larvae: Larval density adjustment.

Black soldier fly (Hermetia illucens) larvae (BSFL) are common insects that are known for bioconversion of organic waste into a sustainable utilization resource. However, a strategy to increase antibiotic resistance gene (ARG) elimination in sustainable and economic ways through BSFL is lacking. In the present study, different larval densities were employed to assess the mcr-1 and tetX elimination abilities, and potential mechanisms were investigated. The application and economic value of each larval density were also analyzed. The results showed that the 100 larvae cultured in 100 g of manure group had the best density because the comprehensive disadvantage evaluation ratio was the lowest (14.97%, good bioconversion manure quality, low ARG deposition risk and reasonable larvae input cost). Further investigation showed that mcr-1 could be significantly decreased by BSFL bioconversion (4.42 ×107 copies/g reduced to 4.79 ×106-2.14 ×105 copies/g)(P＜0.05); however, mcr-1 was increasingly deposited in the larval gut with increasing larval density. The tetX abundance was stabilized by BSFL bioconversion, except that the abundance at the lowest larval density increased (1.22 ×1010 copies/g increase, 34-fold). Escherichia was the host of mcr-1 and tetX in all samples, especially in fresh manure; Alcaligenes was the host of tetX in bioconversion manure; and the abundance of Alcaligenes was highly correlated with the pH of bioconversion manure. The pH of bioconversion manure was extremely correlated with the density of larvae. Klebsiella and Providencia were both hosts of tetX in the BSF larval gut, and Providencia was also the host of mcr-1 in the BSF larval gut. The density of larvae influenced the bioconversion manure quality and caused the ARG host abundance to change to control the abundance of ARGs, suggesting that larval density adjustment was a useful strategy to manage the ARG risk during BSFL manure bioconversion.

C/N-Dependent Element Bioconversion Efficiency and Antimicrobial Protein Expression in Food Waste Treatment by Black Soldier Fly Larvae.

The black soldier fly (BSF), Hermetia illucens, has emerged as a promising species for waste bioconversion and source of antimicrobial proteins (AMPs). However, there is a scarcity of research on the element transformation efficiency and molecular characterization of AMPs derived from waste management. Here, food waste treatment was performed using BSF larvae (BSFL) in a C/N ratio of 21:1−10:1, with a focus on the C/N-dependent element bioconversion, AMP antimicrobial activity, and transcriptome profiling. The C-larvae transformation rates were found to be similar among C/Ns (27.0−35.5%, p = 0.109), while the N-larvae rates were different (p = 0.001), with C/N 21:1−16:1 (63.5−75.0%) being higher than C/N 14:1−10:1 (35.0−45.7%). The C/N ratio did not alter the antimicrobial spectrum of AMPs, but did affect the activities, with C/N 21:1 being significantly lower than C/N 18:1−10:1. The lysozyme genes were found to be significantly more highly expressed than the cecropin, defensin, and attacin genes in the AMP gene family. Out of 51 lysozyme genes, C/N 18:1 and C/N 16:1 up-regulated (p < 0.05) 14 and 12 genes compared with C/N 21:1, respectively, corresponding to the higher activity of AMPs. Overall, the element bioconversion efficiency and AMP expression can be enhanced through C/N ratio manipulation, and the C/N-dependent transcriptome regulation is the driving force of the AMP difference.

Effects of three major nutrient contents, compost thickness and treatment time on larval weight, process performance and residue component in black soldier fly larvae (Hermetia illucens) composting.

The cellulose content in vegetable waste (VW) is high and cannot be directly digested by black soldier fly larvae (BSFL). In this study, in order to treat VW using BSFL composting, kitchen waste (KW) is used as the only nutritional supplement for VW to analyze the effects of the different contents of crude protein (CP), crude fat (EE), carbohydrate (3C), compost thickness (CT), and treatment time on the larval weight (LW), survival rate (SR), dry matter reduction rate (DMR), bioconversion rate (BCR), physical and chemical properties of BSFL sand and changes in the microbial community. Our results showed that when the average 3C content increased by 40%, the average LW increased by 47.6%, and the SR, DMR, BCR, and organic matter (OM) content increased by 16.82%, 8.5%, 4.77%, and 3.86%, respectively. In contrast, when the average compost thickness increased by 5 cm, the average weight of BSFL decreased by 22.64%, while the SR of larvae, DMR, BCR, OM, and total nutrients (TN + P2O5 + K2O) decreased by 5%, 5.2%, 4.42%, 9.6%, and 0.78%, respectively. Germination test showed that BSFL sand alone could not be used as soilless culture substrate. After BSFL treatment, we found that the dominant phyla in BSFL sand were Firmicutes (95.77%), Proteobacteria (2.54%), Actinobacteria (0.74%), and Chloroflexi (0.6%). Our results indicate that BSFL composting is an effective method of treating VW, and 3C content and CT have a significant effect on BSFL composting.

Black soldier fly larvae for treatment and segregation of commingled municipal solid waste at different environmental conditions.

The commingled nature of the municipal solid waste in many developing nations is one of the primary reasons behind the failure to its successful management. Although there are technologies to effectively treat and process well-segregated and classified waste, they are ineffective in managing the commingled waste. Commingled waste has neither calorific value enough to support waste-to-energy operations nor is it suitable for producing quality compost to generate market value. In this article, a novel Black Soldier Fly Larva (BSFL) -based technology has been proposed and tested to auto-segregate and treat the biodegradable fraction of the Commingled Municipal Solid Waste (CMSW). Several BSFL feeding experiments on five different CMSW compositions were conducted at various temperatures and relative humidity conditions. BSFL could selectively consume the biodegradable fraction of the CMSW to convert it into its body mass and separate itself from the rest of the waste, which mainly consisted of inert and recyclable waste fractions that can be further treated using appropriate waste treatment technology. The mature larvae or the pre-pupae grown at the expense of the biodegradable waste fraction can find several commercial uses. The larvae's consumption rate and weight gain were dependent on the environmental conditions; 30 °C and relative humidity of 65-75% provided the highest consumption rate and most significant weight gain. The batch experiments in the laboratory proved that BSFL could be promising for the treatment of CMSW. The experimental data presented may help design a process for further scaling up an effective treatment method for CMSW, which might benefit many developing nations in managing their waste effectively and economically.

Cardboard supplementation on the growth and nutritional content of black solider fly (Hermetia illucens) larvae and resulting frass.

A 10-day trial was conducted to compare the production and fatty acid composition of black soldier fly (Hermetia illucens) larvae (BSFL) when grown without or with cardboard supplementation at 2.2% on a dry weight basis. The final biomass of BSFL or waste reduction was not significantly impacted by cardboard. The fatty acids of C10 and C22:6n-3 were significantly higher in BSFL in the cardboard treatment, but crude lipid significantly reduced. The leftover BSFL frass had significantly higher sulfur, zinc, manganese and boron at the expense of lower nitrogen (91.2% versus 8.73% in control versus cardboard, respectively). These preliminary results appear to indicate that the growth and nutritional value of BSFL were not adversely compromised while the frass can be enhanced by adding relatively small amounts (2.2%) of cardboard. Further studies could be conducted to investigate the implications of higher inclusion levels.

Black soldier fly (Hermetia illucens) larvae meal in diets of European seabass: Effects on antioxidative capacity, non-specific immunity, transcriptomic responses, and resistance to the challenge with Vibrio alginolyticus.

Black soldier fly larvae meal (BSFLM) has been successfully demonstrated as a promising fish meal (FM) replacer in diets of several fish species including European seabass (Dicentrarchus labrax). However, its impacts on antioxidant capacity, and immune responses of treated fish are still poorly understood. A 60-day feeding trial was carried out to evaluate the effects of partial substitution of FM with different levels of dry BSFLM on the antioxidative status, non-specific immunity, transcriptomic responses, and resistance of European seabass to the challenge with Vibrio alginolyticus. Four isoproteic (45%) and isolipidic diets were formulated by replacing 0.0%, 25%, 35%, and 50% of the dietary FM. Each diet was randomly assigned to four fish groups (in triplicates) (initial mean body weight, 12.1 ± 0.21 g) (20 fish per aquarium) (n = 240). Fish were fed three times daily to the apparent satiation. At the end of the feeding trial, serum antioxidant biomarkers such as malondialdehyde levels, and catalase, superoxide dismutase, and glutathione peroxidase enzyme activities were significantly increased in all BSFLM groups in a dose-dependent manner compared to the control group (P < 0.05). The non-specific immune indices, including phagocytic activity, phagocytic index, serum lysozyme and respiratory burst activities were significantly elevated in BSFLM groups compared to those in the control group (P < 0.05). Significant upregulation of the mRNA expression levels of hepatic heat shock protein 70, interleukin-1beta and interleukin-10 genes were observed in all BSFLM groups compared to the control group (P < 0.05). Additionally, after the challenge with V. alginolyticus, the relative percent of survival was significantly elevated in fish groups fed on diets containing graded levels of BSFLM over the control group (P < 0.05). Conclusively, the present study suggests the potential efficacy of partial replacement of dietary FM protein for up to 50% by BSFLM without negative effects on fish health with possible potentiation of the antioxidative status, and the immune responses of the European seabass.

Insect (black soldier fly, Hermetia illucens) meal supplementation prevents the soybean meal-induced intestinal enteritis in rainbow trout and health benefits of using insect oil.

Black solider fly larvae (BSFL) and their oils (BSFLO) are receiving increasing attention as sustainable ingredients in fish feeds, but mostly as replacements to marine sources. There were two aims to this study; in exp. 1, soybean meal (SBM)-based diets were formulated to contain BSFL as supplements at 0 (SBM), 8 (SBM + BSFLlow) or 16% (SBM + BSFLhigh) with a control diet being fishmeal-based (FM). In exp. 2, diets included only fish oil (FO), soybean oil (SBO), BSFLO or BSFLO + bile acid (BA), and all lipid sources were added at 16%. Both experiments were run at the same time and fed to rainbow trout (32 g) with each treatment being triplicated. After 10 weeks the fish were sampled for liver and distal intestine histology, expression of genes responsible for inflammation in the intestine and kidneys, and serum peroxidase and lysozyme activities. In exp. 1, supplementations of BSFL effectively prevented SBM-induced intestinal enteritis, down-regulated intestinal prostaglandin and interferon regulatory factor 1 (IRF-1), while the SBM + BSFLhigh diet significantly increased serum lysozyme activity. In exp. 2, BSFLO caused no histomorphological change to the liver or intestine, but kidney interluekin-8, tumor necrosis factor and IRF-1 were significantly upregulated along with significantly higher serum peroxidase activity. The inclusion of BA in the BSFLO diets significantly upregulated intestinal prostaglandin gene expression. Overall, BSFL supplementations of 8 or 16% prevented SBM-induced intestinal enteritis based on histological observations, which was supported by a down-regulation in pro-inflammatory genes and enhanced innate immunity. Meanwhile, the use of BSFLO showed some immunological benefits. Therefore, these sustainable resources are recommended in the diets of rainbow trout, especially when using elevated levels of plant-based proteins.