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.

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.

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.

Inhibition of pathogenic microorganisms in solid organic waste via black soldier fly larvae-mediated management.

Inadequately managed solid organic waste generation poses a threat to the environment and human health globally. Biotransformation with the black soldier fly larvae (BSFL) is emerging as talent technology for solid waste management. However, there is a lack of understanding of whether BSFL can effectively suppress potential pathogenic microorganisms during management and the underlying mechanisms. In this study, we investigated the temporal variations of microorganisms in two common types of solid waste, i.e., kitchen waste (KW) and pig manure (PM). Natural composting and composting with BSFL under three different pH levels (pH 5, 7, and 9) were established to explore their impact on microbial communities in compost and the gut of BSFL. The results showed that the compost of kitchen waste and pig manure led to an increase in relative abundance of various potentially pathogenic bacteria. Temporal gradient analyses revealed that the most substantial reduction in the relative abundance and diversity of potentially pathogenic microorganisms occurred when the initial pH of both two wastes were adjusted to 7 upon the introduction of BSFL. Through network and pls-pm analysis, it was discovered that the gut microbiota of BSFL occupied an ecological niche in the compost, inhibiting the proliferation of potentially pathogenic microorganisms. This study has revealed the potential of BSFL in reducing public health risks during the solid waste management process, providing robust support for sustainable waste management.

Evolution of the Black solider fly larvae gut antibiotic resistome during kitchen waste disposal.

Kitchen waste (KW) is an important reservoir of antibiotic resistance genes (ARGs). Black solider fly larvae (BSFL) are extensively employed in KW disposal, closely linking to their robust gut microbes. However, antibiotic resistome in BSFL gut during the KW disposal processes and the mechanism remain unclear. In the present study, the antibiotic resistome in BSFL gut within the 12 days KW disposal processes were investigated. Results showed that, ARGs abundance initially increased and subsequently decreased, the five most prevalent core ARG classes were tetracycline, aminoglycoside, cephalosporin, lincosamide and multidrug. A total of 7 MGE types were observed and the horizontal gene transfer (HGT) of ARGs was predominantly mediated by plasmids. Host microbes were mainly categorized into Proteobacteria (98.12 %) and their assemblies were mainly classified into the deterministic processes. To elucidate the driving mechanisms, the mantel test and the structural equation model (SEM) were developed. Results indicated that microbial functions (0.912, p < 0.0001) and microbial community (1.014, p = 0.036), consistently showed very significant relationships with the patterns of ARGs, which presented higher direct effects than indirect effects. Overall, this study makes an initial contribution to a more deepgoing comprehension of the gut antibiotic resistome of BSFL during KW disposal.

Black soldier fly (Diptera: Stratiomyidae) larvae reduce cyathostomin (Nematoda: Strongylidae) eggs but develop poorly on horse manure.

Cyathostomins are common digestive tract parasites of grazing horses that spread through contact with horse feces. Horse feces are colonized by a variety of organisms, some of which could serve to reduce parasite loads in horse pastures. Black soldier fly (Hermetia illucens L.; Diptera: Stratiomyidae) larvae (BSFL) could be an ideal candidate for biological control of cyathostomins, due to their near-global distribution, low risk of pathogen transmission, ability to develop on a variety of nutrient-poor substrates (including horse manure), and dramatic effect on microbial communities that cyathostomins depend on. Here, using controlled feeding bioassays, we evaluated the effect of BSFL on cyathostomin egg densities in horse manure while also tracking BSFL performance on manure relative to standard grain-based diets. We found that BSFL consumed less substrate, were slower to reach the prepupal stage, and ultimately yielded less biomass when reared on horse manure compared to grain-based diets. However, BSFL reduced average cyathostomin egg densities in horse manure by over 3-fold. Overall, our results suggest that despite horse manure being a poor substrate for BSFL growth, BSFL effectively reduce cyathostomin egg loads in infected horse manure, though the mechanisms by which they do this are uncertain. While BSFL are known to transform the microbial communities within a diversity of rearing substrates, their effect on larger, parasitic organisms in animal manures may be underappreciated. Promoting the decomposition of infected horse manure with BSFL might be a promising approach to managing parasite populations among grazing horses.

Comparative Metagenomic and Metatranscriptomic Analyses Reveal the Response of Black Soldier Fly (Hermetia illucens) Larvae Intestinal Microbes and Reduction Mechanisms to High Concentrations of Tetracycline.

Black soldier fly (Hermetia illucens L) larvae (BSFL) possess remarkable antibiotic degradation abilities due to their robust intestinal microbiota. However, the response mechanism of BSFL intestinal microbes to the high concentration of antibiotic stress remains unclear. In this study, we investigated the shift in BSFL gut microbiome and the functional genes that respond to 1250 mg/kg of tetracycline via metagenomic and metatranscriptomic analysis, respectively. The bio-physiological phenotypes showed that the survival rate of BSFL was not affected by tetracycline, while the biomass and substrate consumption of BSFL was slightly reduced. Natural BSFL achieved a 20% higher tetracycline degradation rate than the germ-free BSFL after 8 days of rearing. Metagenomic and metatranscriptomic sequencing results revealed the differences between the entire and active microbiome. Metatranscriptomic analysis indicated that Enterococcus, Vagococcus, Providencia, and Paenalcaligenes were the active genera that responded to tetracycline. Furthermore, based on the active functional genes that responded to tetracycline pressure, the response mechanisms of BSFL intestinal microbes were speculated as follows: the Tet family that mediates the expression of efflux pumps expel tetracycline out of the microbes, while tetM and tetW release it from the ribosome. Eventually, tetracycline was degraded by deacetylases and novel enzymes. Overall, this study provides novel insights about the active intestinal microbes and their functional genes in insects responding to the high concentration of antibiotics.

Synergistic bioconversion of organic waste by black soldier fly (Hermetia illucens) larvae and thermophilic cellulose-degrading bacteria.

Introduction: This study examines the optimum conversion of Wuzhishan pig manure by Black Soldier Fly Larvae (BSFL) at various phases of development, as well as the impact of gut microbiota on conversion efficiency. Method and results: In terms of conversion efficiency, BSFL outperformed the growing pig stage (GP) group, with significantly higher survival rates (96.75%), fresh weight (0.23 g), and larval conversion rate (19.96%) compared to the other groups. Notably, the GP group showed significant dry matter reductions (43.27%) and improved feed conversion rates (2.17). Nutritional composition varied, with the GP group having a lower organic carbon content. High throughput 16S rRNA sequencing revealed unique profiles, with the GP group exhibiting an excess of Lactobacillus and Clostridium. Promising cellulose-degrading bacteria in pig manure and BSFL intestines, including Bacillus cereus and Bacillus subtilis, showed superior cellulose degradation capabilities. The synergy of these thermophilic bacteria with BSFL greatly increased conversion efficiency. The BSFL1-10 group demonstrated high growth and conversion efficiency under specific conditions, with remarkable larval moisture content (71.11%), residual moisture content (63.20%), and waste reduction rate (42.28%). Discussion: This study sheds light on the optimal stages for BSFL conversion of pig manure, gut microbiota dynamics, promising thermophilic cellulose-degrading bacteria, and the significant enhancement of efficiency through synergistic interactions. These findings hold great potential for sustainable waste management and efficient biomass conversion, contributing to environmental preservation and resource recovery.

Food industry waste - An opportunity for black soldier fly larvae protein production in Tanzania.

Black soldier fly larvae composting is an emerging treatment option with potential to improve biowaste valorization in cities of low-income countries. This study surveyed the current generation and management status of food industry biowaste and their availability and suitability as potential feedstock for black soldier fly larvae (BSFL) composting treatment in three Tanzania cities, Dar es Salaam, Mwanza, and Dodoma. Biowaste-generating food industry companies (n = 29) in the three cities were found to produce banana peels, mango seeds, sunflower press cake, brewery waste, and coffee husks in large quantities (~100,000-1,000,000 kg y-1). Around 50 % of these companies (16/29), primarily vegetable oil companies (10/11), either sold or gave away their waste as animal feed, while most companies (9/11) with unutilized food industry waste landfilled the generated biowaste. Multi-criteria analysis based on substrate availability criteria identified banana peels, mango seeds, and coffee husks with total score points of ≥10/12 as the most suitable feedstock for BSFL composting. However, multi-criteria analysis based on physical-chemical criteria identified brewery waste and sunflower press cake with total score points of ≥11/15 as the most suitable feedstock. Combined availability and physical-chemical properties of individual biowastes showed that all identified types of food industry biowaste can be suitable feedstock for producing BSFL biomass for protein production, but certain waste streams needed to be mixed with other waste streams prior to BSFL-composting to ensure sufficient availability and provide a balanced nutritional profile compared with the single-source biowastes. This study concluded that large volumes of food industry waste are being generated from food industry companies in Tanzania and there is need to establish new biowaste management interventions for resource recovery. Furthermore, for interested stakeholders in the waste management business, multi-stream BSFL-composting can be a suitable solution for managing and closing nutrient loops of the unutilized food industry biowaste in Tanzania and in other similar settings globally.

Performance and emission characteristics of a diesel engine fuelled by biodiesel from black soldier fly larvae: Effects of synthesizing catalysts with citric acid.

Biodiesel has several environmental benefits, such as biodegradability, renewability and lower soot emissions. However, biodiesel has undesirable properties such as higher viscosity and density and low calorific value compared to petroleum diesel, resulting in high Brake Specific Fuel Consumption (BSFC), reduced Brake Power (BP) and increased NOX emissions creating an environmental concerns in biodiesel development. This study investigated the effects of synthesizing transesterification catalysts (CaO and NaOH) with Citric Acid (CA) on the quality of biodiesel and biodiesel blends produced from Black Soldier Fly Larvae (BSFL) (Hermetia Illucens). The quality of biodiesel and blends was determined based on fuel properties, engine performance and emission composition characteristics. The tests were performed on a single-cylinder, four-stroke, Compression Ignition (CI) diesel engine at five loads at a constant speed of 1500 rpm. The results showed that synthesizing the catalysts with CA significantly affected the fatty acid profile of the biodiesel compared to physical fuel properties. B100 (pure BSFL biodiesel) exhibited higher BSFC by 10.57-13.97 % and lower BP by 4.21-7.83 % than diesel fuel. However, the Brake Thermal Efficiency (BTE) of biodiesel was higher than that of diesel fuel by 0.82-4.34 % at maximum load. Synthesizing catalysts with CA improved the viscosity of biodiesel by 0.93-2.81 % and effectively reduced NOX, HC and Smoke opacity by 2.23-3.16 %, 4.95-5.83 % and 20.51-41.15 %, respectively.

Effects of further composting black soldier fly larvae manure on toxic metals and resistant bacteria communities by cornstalk amendment.

Rapid composting by black soldier fly larvae (BSFL) may be insufficient to maturation and humification of composting and further composting is necessary. The purpose of this study was to explore cornstalk addition on toxic metals (Cu, Zn, Pb and Cd), toxic metals resistance bacterial (TMRB) destiny and their relationship with physicochemical factors during BSFL manure composting. High-throughput sequencing was performed by six treatments, namely T1 to T6, where T1 to T3 were BSFL manures from chicken, pig and dairy manure, respectively, and T4 to T6 were same manures and utilized cornstalk to adjust C/N to 25. The results showed that cornstalk amendment could enhance the toxic metals immobilization rate compared to control treatments in the ultimate product. TMRB indicated that the major potential hosts bacteria were Firmicutes, Bacteroidota, Proteobacteria, Acidobacteriota and Actinobacteriota, and the sum relative abundance were 63.33%, 90.62%, 83.62%, 69.38%, 50.66% and 90.52% in T1 to T6 at the end of composting. Bacteria diversity and heat map revealed composting micro-ecology with additive cornstalk to remarkably effect main resistant bacterial distribution via adjusting environmental factors and potential hosts bacterial. Finally, T5 treatment was able to greatly decrease the TMRB abundance, and improve the ability of composting and ultimate product quality.

Evaluation of Black Soldier Fly larvae (Hermetia illucens) as a protein supplement for beef steers consuming low-quality forage.

Black Soldier Fly larvae (BSFL; Hermetia illucens) has been the focus of recent feeding trials in poultry, swine, and fish; however, in vivo research has not yet been conducted in cattle. Accordingly, a study was conducted to evaluate the effects of BSFL as a protein supplement in beef steers. Six steers (603 ± 20 kg of BW, n = 3 and 404 ± 17 kg of BW, n = 3) consuming King Ranch bluestem hay (6.55% dry matter [DM] crude protein [CP]) ad libitum were used in two simultaneous 3 × 3 Latin squares. Steers were provided one of three treatments each period: 1) a control with no supplement (CON), 2) a supplement comprised of conventional feed ingredients with whole cottonseed and soybean meal as the main protein sources (CONV), and 3) a supplement with BSFL as the main protein source (BSFL). Three 14-d periods were conducted with 8 d to adapt to treatments, 5 d to measure intake and digestion, and 1 d to complete a ruminal fermentation profile. DM, organic matter (OM), CP, neutral detergent fiber (NDF), and acid detergent insoluble ash were determined in forage, supplement, ort, and fecal samples. Protein supplementation as CONV or BSFL stimulated forage OM intake (P ≤ 0.01) relative to CON with a trend for a difference (P = 0.08) between the supplements such that CONV steers consumed more FOMI than BSFL steers. Treatment affected total digestible OM intake (TDOMI; P ≤ 0.01); TDOMI for CON steers was 47.5 g/kg metabolic body weight (MBW) which was significantly less (P ≤ 0.01) than that of CONV or BSFL steers. Steers supplemented with CONV consumed significantly more TDOMI than BSFL steers (P = 0.05; 62.2 vs. 60.1 g/kg MBW, respectively). Treatment did not significantly affect digestibility of DM, OM, or NDF (P > 0.32). There was also not a significant effect (P ≥ 0.17) of treatment on ruminal ammonia-N, total volatile fatty acids, or ruminal pH. Overall, these data indicate that BSFL may be an effective protein supplement for beef cattle consuming low-quality forage.

Assessing the Effectiveness of Providing Live Black Soldier Fly Larvae (Hermetia illucens) to Ease the Weaning Transition of Piglets.

Weaning is a stressful event for piglets, involving substantial changes to their nutritional and social environment. Providing edible enrichment around weaning may ease the weaning transition by increasing pre-weaning feed intake and improving post-weaning performance, health, behavior, and affective state. In this study, we investigated the effects of providing live black soldier fly larvae (BSFL) as edible enrichment pre- and/or post-weaning. Pre-weaning, piglets received either only creep feed (Pre-C, n = 14 litters) or creep feed and live BSFL (Pre-L, n = 15 litters) ad libitum, and post-weaning piglets either had no access to live BSFL (Post-C, n = 24 pens) or they could rotate tubes that released BSFL (Post-L, n = 24 pens) at levels up to 20% of their expected daily dry matter intake, resulting in treatments CC, CL, LC, and LL. No interaction between pre- and post-weaning treatment was found for any of the measured parameters. Before weaning, Pre-L piglets preferred to interact with larvae over creep feed, and Pre-C piglets interacted more with creep feed than Pre-L piglets. Total time spent on feed-directed behaviors did not differ. Continuous larvae provisioning increased caecum length and proximal stomach digesta pH, while it decreased the passage of glucose and fluorescein isothiocyanate through the colon wall on d3 post-weaning (CC vs. LL, n = 12 piglets/treatment). Post-weaning diarrhea and final body weight were not affected by treatment. After weaning, Pre-C piglets tended to eat more and grew marginally faster than Pre-L piglets. Post-C piglets spent more time eating and had a higher feed intake post-weaning than Post-L piglets. Based on home-pen behavioral observations, Post-L piglets actively explored and ate the larvae. Post-C piglets spent more time on exploring the environment and nosing pen mates, and they spent more time on manipulating pen mates on d8 and played more on d8 & 15 compared to Post-L piglets. Piglet responses to a novel environment and an attention bias test on d4 & 5 post-weaning were not influenced by larvae provisioning. In conclusion, pre-weaning larvae provisioning did not improve pre-weaning feed intake and post-weaning performance, however post-weaning larvae provisioning did benefit piglet behavior as less manipulation of pen mates was observed.

Black soldier fly larvae (BSFL) and their affinity for organic waste processing.

There are two major problems that we are facing currently. Firstly, a growing human population continues to contribute to the increased food demand. Secondly, the volume of organic waste produced will threaten human health and the quality of the environment. Recently, there is an increasing number of efforts placed into farming insect biomass to produce alternative feed ingredients. Black soldier fly larvae (BSFL), Hermetia illucens have proven to convert organic waste into high-quality nutrients for pet foods, fish and poultry feeds, as well as residue fertilizer for soil amendment. However, better BSFL feed formulation and feeding approaches are necessary for yielding a higher nutrient content of the insect body, and if performed efficiently, whilst converting waste into higher value biomass. Lastly, this paper reveals that BSFL, in fact, thrives in various ranges of organic matter composition and with simple rearing systems.

Physico-chemical and colloidal properties of protein extracted from black soldier fly (Hermetia illucens) larvae.

The black soldier fly larvae (BSFL), Hermetia illucens (Linnaeus), has been largely utilized for animal feed. Due to its interesting composition, BSFL has great potential to be further implemented in the human diet. Herein we compared the flour and protein extract composition based on their moisture, ash, amino acids, mineral, and protein content. To have wide knowledge on protein profile and behavior, SDS-page electrophoresis, Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) were used to give information about protein structure and thermal stability, respectively. The flour and protein extract contained respectively 37.3% and 61.1% of protein. DSC graph reported a glass transition temperature around 30 °C, recognizable by a shift in the curve, and an endothermic peak for solid melting at around 200 °C. FTIR analysis showed the main amide bands (A, B, I, II, III) for the flour and protein extract. The foam properties of BSFL protein extract were explored under different temperatures treatment, and the best foam stability was reached at 85 °C with 15 min of treatment. The data highlight the promising techno-functional properties of BSFL protein extract, and that the nutritional composition might be suitable for further use of BSFL as food fortification system.

A Molecular Survey of Bacterial Species in the Guts of Black Soldier Fly Larvae (Hermetia illucens) Reared on Two Urban Organic Waste Streams in Kenya.

Globally, the expansion of livestock and fisheries production is severely constrained due to the increasing costs and ecological footprint of feed constituents. The utilization of black soldier fly (BSF) as an alternative protein ingredient to fishmeal and soybean in animal feed has been widely documented. The black soldier fly larvae (BSFL) used are known to voraciously feed and grow in contaminated organic wastes. Thus, several concerns about their safety for inclusion into animal feed remain largely unaddressed. This study evaluated both culture-dependent sequence-based and 16S rDNA amplification analysis to isolate and identify bacterial species associated with BSFL fed on chicken manure (CM) and kitchen waste (KW). The bacteria species from the CM and KW were also isolated and investigated. Results from the culture-dependent isolation strategies revealed that Providencia sp. was the most dominant bacterial species detected from the guts of BSFL reared on CM and KW. Morganella sp. and Brevibacterium sp. were detected in CM, while Staphylococcus sp. and Bordetella sp. were specific to KW. However, metagenomic studies showed that Providencia and Bordetella were the dominant genera observed in BSFL gut and processed waste substrates. Pseudomonas and Comamonas were recorded in the raw waste substrates. The diversity of bacterial genera recorded from the fresh rearing substrates was significantly higher compared to the diversity observed in the gut of the BSFL and BSF frass (leftovers of the rearing substrates). These findings demonstrate that the presence and abundance of microbiota in BSFL and their associated waste vary considerably. However, the presence of clinically pathogenic strains of bacteria in the gut of BSFL fed both substrates highlight the biosafety risk of potential vertical transmission that might occur, if appropriate pre-and-postharvest measures are not enforced.

Aflatoxin B1 Conversion by Black Soldier Fly (Hermetia illucens) Larval Enzyme Extracts.

The larvae of the black soldier fly (Hermetia illucens L., BSFL) have received increased industrial interest as a novel protein source for food and feed. Previous research has found that insects, including BSFL, are capable of metabolically converting aflatoxin B1 (AFB1), but recovery of total AFB1 is less than 20% when accounting for its conversion to most known metabolites. The aim of this study was to examine the conversion of AFB1 by S9 extracts of BSFL reared on substrates with or without AFB1. Liver S9 of Aroclor-induced rats was used as a reference. To investigate whether cytochrome P450 enzymes are involved in the conversion of AFB1, the inhibitor piperonyl butoxide (PBO) was tested in a number of treatments. The results showed that approximately 60% of AFB1 was converted to aflatoxicol and aflatoxin P1. The remaining 40% of AFB1 was not converted. Cytochrome P450s were indeed responsible for metabolic conversion of AFB1 into AFP1, and a cytoplasmic reductase was most likely responsible for conversion of AFB1 into aflatoxicol.