Black soldier fly larvae meal as a potential modulator of immune, inflammatory, and antioxidant status in gilthead seabream juveniles.

This study aimed to evaluate the effects of fish meal (FM) replacement with defatted Hermetia illucens larvae meal (HM) on the hematological profile, immune parameters, intestinal inflammatory status, and antioxidant response in gilthead seabream juveniles. Four diets were formulated, replacing FM with HM at 0%, 22%, 60%, and 100% levels, corresponding to an inclusion level of 15 (diet HM15), 30 (diet HM30), and 45% (diet HM45), respectively. Over 67 days, fish were fed these diets until apparent visual satiation. Results showed no significant differences in immune parameters or hematological profiles, except for a decrease in hemoglobin and hematocrit levels. In the liver, glucose-6-phosphate dehydrogenase and glutathione peroxidase decreased linearly with HM content, especially at 100% replacement. Glutathione reductase activity was also reduced with HM inclusion, being lower in fish fed diet HM30 compared to the control. Fish fed diet HM15 showed lower hepatic superoxide dismutase activity, while catalase activity and lipid peroxidation remained unaffected. In the intestine, antioxidant enzyme activity was not influenced by HM, but lipid peroxidation linearly decreased with HM inclusion, being lower in the HM30 diet compared to the control. The inclusion of HM reduced the expression of intestinal pro-inflammatory genes (interleukin-1ß and cyclooxygenase-2) while the expression of transforming growth factor ß was higher in fish fed diet HM30 compared to the control and HM45 diets. In conclusion, up to 45% dietary inclusion of HM showed no adverse effects, improving liver antioxidant status, reducing intestinal oxidative stress, and regulating inflammatory gene expression.

Physicochemical, functional, and antioxidant properties of black soldier fly larvae protein.

This study explores the multifaceted attributes of black soldier fly larvae protein (BSFLP), focusing on its physicochemical, functional, and antioxidant properties. BSFLP is characterized by 16 amino acids, with a predominant random coil secondary structure revealed by circular dichroism spectra. Differential scanning calorimetry indicates a substantial thermal denaturation temperature of 97.63°C. The protein exhibits commendable solubility, emulsification, and foaming properties in alkaline and low-salt environments, albeit with reduced water-holding capacity and foam stability under elevated alkaline and high-temperature conditions. In vitro assessments demonstrate that BSFLP displays robust scavenging proficiency against 2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), and hydroxyl radicals, with calculated EC50 values of 1.90 ± 0.57, 0.55 ± 0.01, and 1.14 ± 0.02 mg/mL, respectively, along with notable reducing capabilities. Results from in vivo antioxidant experiments reveal that BSFLP, administered at doses of 300 and 500 mg/kg, significantly enhances the activities of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) (p < 0.05) while simultaneously reducing malondialdehyde levels in both serum and tissues of d-galactose-induced oxidative stress in mice. Moreover, the protein effectively attenuates oxidative damage in liver and hippocampal tissues. These findings underscore the potential utility of BSFLP as a natural antioxidant source, with applications spanning the food, pharmaceutical, and cosmetic industries. PRACTICAL APPLICATION: Black soldier fly larvae protein emerges as an environmentally sustainable reservoir of natural antioxidants, holding significant promise for the food, pharmaceutical, and cosmetic sectors. Its advantageous amino acid composition, robust thermal resilience, and impressive functional attributes position it as a compelling subject for continued investigation and advancement in various applications.

Fat from Hermetia illucens Alters the Cecal Gut Microbiome and Lowers Hepatic Triglyceride Concentration in Comparison to Palm Oil in Obese Zucker Rats.

BACKGROUND: Palm oil (PO) is the most widely utilized plant oil for food production. Owing to the great ecologic problems associated with PO production, sustainably produced fats, such as insect fat, might be a suitable alternative. OBJECTIVES: The hypothesis was tested that fat from Hermetia illucens larvae (HF) compared with PO and soybean oil (SO) has no adverse effects on hepatic lipid metabolism, plasma metabolome, and cecal microbiome in obese Zucker rats. METHODS: Thirty male obese Zucker rats were randomly assigned to 3 groups (SO, PO, HF; n = 10 rats/group) and fed 3 different semisynthetic diets containing either SO, PO, or HF as the main fat source for 4 wk. The effects were evaluated by measurement of liver and plasma lipid concentrations, liver transcriptomics, targeted plasma metabolomics, and cecal microbiomics. RESULTS: Supplementation of HF reduced hepatic triglyceride concentration and messenger ribonucleic acid concentrations of selected genes involved in fatty acid and triglyceride synthesis in comparison to PO (P < 0.05). Pairwise comparison of the Simpson index and Jaccard index showed a higher cecal microbial α- and ß-diversity in rats fed the HF diet than in rats fed the PO diet (P = 0.015 and P = 0.027), but no difference between rats fed the diets with SO or PO. Taxonomic analysis of the cecal microbial community revealed a lower abundance of Clostridium_sensu_stricto_1 and a higher abundance of Blautia, Mucispirillum, Anaerotruncus, Harryflintia, and Peptococcus in rats supplemented with HF than in rats supplemented with PO (P < 0.05). CONCLUSIONS: HF, compared with PO, has liver lipid-lowering effects in obese Zucker rats, which may be caused by a shift in the gut microbial community. Thus, HF might serve as a sustainably produced fat alternative to PO for food production.

Identification of functional microflora underlying the biodegradation of sulfadiazine-contaminated substrates by Hermetia illucens.

The increasing discharge of antibiotic residues into the natural environment, stemming from both human activities and animal farming, has detrimental effects on natural ecosystems and serves as a significant driving force for the spread of antibiotic resistance. Biodegradation is an important method for the elimination of antibiotics from contaminated substrates, but the identifying in situ microbial populations involved in antibiotic degradation is challenging. Here, DNA stable isotope probing (DNA-SIP) was employed to identify active sulfadiazine (SDZ) degrading microbes in the gut of black soldier fly larvae (BSFLs). At an initial SDZ concentration of 100 mg kg-1, the highest degradation efficiency reached 73.99% after 6 days at 28 °C. DNA-SIP revealed the incorporation of 13C6 from labeled SDZ in 9 genera, namely, Clostridum sensu stricto 1, Nesterenkonia, Bacillus, Halomonas, Dysgonomonas, Caldalkalibacillus, Enterococcus, g_unclassified_f_Xanthomonadaceae and g_unclassified_f_Micrococcaceae. Co-occurrence network analysis revealed that a significant positive correlation existed among SDZ degrading microbes in the gut microbiota, e.g., between Clostridium sensu stricto 1 and Nesterenkonia. Significant increases in carbohydrate metabolism, membrane transport and translation were crucial in the biodegradation of SDZ in the BSFL gut. These results elucidate the structure of SDZ-degrading microbial communities in the BSFL gut and in situ degradation mechanisms.

In-depth biological characterization of two black soldier fly anti-Pseudomonas peptides reveals LPS-binding and immunomodulating effects.

As effector molecules of the innate immune system, antimicrobial peptides (AMPs) have gathered substantial interest as a potential future generation of antibiotics. Here, we demonstrate the anti-Pseudomonas activity and lipopolysaccharide (LPS)-binding ability of HC1 and HC10, two cecropin peptides from the black soldier fly (Hermetia Illucens). Both peptides are active against a wide range of Pseudomonas aeruginosa strains, including drug-resistant clinical isolates. Moreover, HC1 and HC10 can bind to lipid A, the toxic center of LPS and reduce the LPS-induced nitric oxide and cytokine production in murine macrophage cells. This suggests that the peptide-LPS binding can also lower the strong inflammatory response associated with P. aeruginosa infections. As the activity of AMPs is often influenced by the presence of salts, we studied the LPS-binding activity of HC1 and HC10 in physiological salt concentrations, revealing a strong decrease in activity. Our research confirmed the early potential of HC1 and HC10 as starting points for anti-Pseudomonas drugs, as well as the need for structural or formulation optimization before further preclinical development can be considered. IMPORTANCE The high mortality and morbidity associated with Pseudomonas aeruginosa infections remain an ongoing challenge in clinical practice that requires urgent action. P. aeruginosa mostly infects immunocompromised individuals, and its prevalence is especially high in urgent care hospital settings. Lipopolysaccharides (LPSs) are outer membrane structures that are responsible for inducing the innate immune cascade upon infection. P. aeruginosa LPS can cause local excessive inflammation, or spread systemically throughout the body, leading to multi-organ failure and septic shock. As antimicrobial resistance rates in P. aeruginosa infections are rising, the research and development of new antimicrobial agents remain indispensable. Especially, antimicrobials that can both kill the bacteria themselves and neutralize their toxins are of great interest in P. aeruginosa research to develop as the next generation of drugs.

The structure of fly Teneurin-m reveals an asymmetric self-assembly that allows expansion into zippers.

Teneurins are conserved cell adhesion molecules essential for embryogenesis and neural development in animals. Key to teneurin function is the ability of its extracellular region to form homophilic interactions in cis and/or in trans. However, our molecular understanding of teneurin homophilic interaction remains largely incomplete. Here, we showed that an extracellular fragment of Teneurin-m, the major teneurin homolog in flies, behaves as a homodimer in solution. The structure of Teneurin-m revealed that the transthyretin-related domain from one protomer and the ß-propeller domain from the other mediates Teneurin-m self-association, which is abolished by point mutation of conserved residues. Strikingly, this architecture generates an asymmetric oligomerization interface that enables expansion of Teneurin-m into long zipper arrays reminiscent of protocadherins. An alternatively spliced site that exists only in vertebrates and regulates homophilic interaction in mammalian teneurins overlaps with the fly Teneurin-m self-association interface. Our work provides a molecular understanding of teneurin homophilic interaction and sheds light on its role in teneurin function throughout evolution.

Exorista sorbillans (Diptera: Tachinidae) parasitism shortens host larvae growth duration by regulating ecdysone and juvenile hormone titers in Bombyx mori (Lepidoptera: Bombycidae).

The tachinid fly, Exorista sorbillans, is a notorious ovolarviparous endoparasitoid of the silkworm, Bombyx mori, causing severe damage to silkworm cocoon industry. Silkworm larvae show typically precocious wandering behavior after being parasitized by E. sorbillans; however, the underlying molecular mechanism remains unexplored. Herein, we investigated the changes in the levels of 20-hydroxyecdysone (20E) and juvenile hormone (JH) titer, and they both increased in the hemolymph of parasitized silkworms. Furthermore, we verified the expression patterns of related genes, which showed an upregulation of 20E signaling and biosynthesis genes but a significant downregulation of ecdysone oxidase (EO), a 20E inactivation enzyme, in parasitized silkworms. In addition, related genes of the JH signaling were activated in parasitized silkworms, while related genes of the JH degradation pathway were suppressed, resulting in an increase in JH titer. Notably, the precocious wandering behavior of parasitized silkworms was partly recoverable by silencing the transcriptions of BmCYP302A1 or BmCYP307A1 genes. Our findings suggest that the developmental duration of silkworm post parasitism could be shortened by regulation of 20E and JH titers, which may help silkworm to resist the E. sorbillans infestation. These findings provide a basis for deeper insight into the interplay between silkworms and E. sorbillans and may serve as a reference for the development of a novel approach to control silkworm myiasis.

Isolated and identified pathogenic bacteria from black soldier fly larvae with "soft rot" reared in mass production facilities and its incidence characteristics.

The black soldier fly larvae (BSFL) can transform organic waste into high-end proteins, lipids, chitin, biodiesel, and melanin at an industrial scale. But scaling up of its production capacity has also posed health risks to the insect itself. In this investigation, larval "soft rot" which is occurring in mass production facilities that cause larval developmental inhibition and a certain degree of death was reported. Responsible pathogen GX6 was isolated from BSFL with "soft rot" and identified to be Paenibacillus thiaminolyticus. No obvious impact on larval growth was observed when treated with GX6 spores, whereas mortality of 6-day-old BSFL increased up to 29.33% ± 2.05% when GX6 vegetative cells (1 × 106 cfu/g) were inoculated into the medium. Moreover, higher temperature further enhanced the BSFL mortality and suppressed larval development, but increasing substrate moisture showed the opposite effect. The middle intestine of infected larvae became swollen and transparent after dissection and examination. Transmission electron microscopy (TEM) observation indicated that GX6 had destroyed the peritrophic matrix and intestinal microvilli and damaged epithelial cells of larval gut. Furthermore, 16S rRNA gene sequencing analysis of intestinal samples revealed that gut microflora composition was significantly altered by GX6 infection as well. It can be noticed that Dysgonomonas, Morganella, Myroides, and Providencia bacteria became more numerous in the intestines of GX6-infected BSFL as compared to controls. This study will lay foundations for efficient control of "soft rot" and promote healthy development of the BSFL industry to contribute to organic waste management and circular economy.

Inoculation with black soldier fly larvae alters the microbiome and volatile organic compound profile of decomposing food waste.

The black soldier fly (BSF; Hermetia illucens) is used in sustainable processing of many types of organic waste. However, organic waste being decomposed by BSF produces strong odors, hindering more widespread application. The odor components and how they are produced have yet to be characterized. We found that digestion of food waste by BSF significantly alters the microbial flora, based on metagenomic analyses, and the odor components generated, as shown by thermal desorption gas chromatography mass spectrometry analysis. Inoculation with BSF significantly decreased production of volatile organic sulfur compounds (dimethyl disulfide and dimethyl trisulfide), which are known to be released during methionine and cysteine metabolism by Lactobacillus and Enterococcus bacteria. BSF inoculation significantly changed the abundance of Lactobacillus and Enterococcus and decreased microbial diversity overall. These findings may help in optimizing use of BSF for deodorization of composting food waste.

Odorant-binding Protein 10 From Bradysia odoriphaga (Diptera: Sciaridae) Binds Volatile Host Plant Compounds.

Bradysia odoriphaga (Diptera: Sciaridae) is a major insect pest of seven plant families including 30 commercial crops in Asia. The long-term use of chemical pesticides leads to problems such as insect resistance, environmental issues, and food contamination. Against this background, a novel pest control method should be developed. In insects, odorant-binding proteins (OBPs) transport odor molecules, including pheromones and plant volatiles, to olfactory receptors. Here, we expressed and characterized the recombinant B. odoriphaga OBP BodoOBP10, observing that it could bind the sulfur-containing compounds diallyl disulfide and methyl allyl disulfide with Ki values of 8.01 µM and 7.00 µM, respectively. Homology modeling showed that the BodoOBP10 3D structure was similar to that of a typical OBP. Both diallyl disulfide and methyl allyl disulfide bound to the same site on BodoOBP10, mediated by interactions with six hydrophobic residues Met70, Ile75, Thr89, Met90, Leu93, and Leu94, and one aromatic residue, Phe143. Furthermore, silencing BodoOBP10 expression via RNAi significantly reduced the electroantennogram (EAG) response to diallyl disulfide and methyl allyl disulfide. These findings suggest that BodoOBP10 should be involved in the recognition and localization of host plants.

Feeding of Hermetia illucens Larvae Meal Attenuates Hepatic Lipid Synthesis and Fatty Liver Development in Obese Zucker Rats.

The present study tested the hypothesis that dietary insect meal from Hermetia illucens (HI) larvae attenuates the development of liver steatosis and hyperlipidemia in the obese Zucker rat. To test the hypothesis, a 4-week trial with male, obese Zucker rats (n = 30) and male, lean Zucker rats (n = 10) was performed. The obese rats were assigned to three obese groups (group O-C, group O-HI25, group O-HI50) of 10 rats each. The lean rats served as a lean control group (L-C). Group L-C and group O-C were fed a control diet with 20% casein as protein source, whereas 25% and 50% of the protein from casein was replaced with protein from HI larvae meal in the diets of group O-HI25 and O-HI50, respectively. The staining of liver sections with Oil red O revealed an excessive lipid accumulation in the liver of group O-C compared to group L-C, whereas liver lipid accumulation in group O-HI25 and O-HI50 was markedly reduced compared to group O-C. Hepatic concentrations of triglycerides, cholesterol, C14:0, C16:0, C16:1, C18:0, C18:1, the sum of total fatty acids and hepatic mRNA levels of several genes associated with lipid synthesis and plasma concentration of cholesterol were markedly higher in group O-C than in group L-C, but lower in group O-HI50 than in group O-C (p < 0.05). In conclusion, partial replacement of casein by HI larvae meal attenuates liver steatosis and dyslipidemia in obese Zucker rats. This suggests that HI larvae meal serves as a functional food protecting from obesity-induced metabolic disorders.

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.

Imaging CaLexA and TRIC Signals in the Dissected Fly Central Nervous System.

Calcium-dependent nuclear import of LexA (CaLexA) and transcriptional reporter of intracellular calcium (TRIC) are transcription-based genetically encoded calcium indicators (transcriptional GECIs). When expressed in neurons, CaLexA and TRIC report neuronal activity by converting intracellular calcium levels into transcription activities and, subsequently, reporter gene expression. CaLexA and TRIC have been used successfully in many studies to label neurons associated with particular behaviors, regulated by internal states, or evoked by specific sensory inputs. This protocol details procedures for generating flies expressing genetic components of CaLexA and TRIC in specific neurons, performing immunostaining to label CaLexA and TRIC signals, and quantifying the signals using an open-source imaging analysis software.

Antimicrobial peptide HI-3 from Hermetia illucens alleviates inflammation in lipopolysaccharide-stimulated RAW264.7 cells via suppression of the nuclear factor kappa-B signaling pathway.

Hermetia illucens-3 (HI-3), an active insect antimicrobial peptide extracted from H. illucens larvae, exerts antibacterial and anticancer activity. However, the inflammatory effects and their relative molecular mechanisms remain unclear. To explore the inflammatory effects of HI-3, an inflammatory model was induced using 1 ng/mL LPS in RAW264.7 cells. The cell viability and phagocytosis of LPS-stimulated RAW264.7 cells were then detected after HI-3 treatment. Furthermore, the antioxidant activity, the levels of proinflammatory cytokines, and the expression levels of both p65 and inhibitor of nuclear factor kappa B (IκB) were measured. Results showed that HI-3 could inhibit the differentiation, proliferation, phagocytosis, and antioxidant ability, as well as the secretion and messenger RNA expression levels of IL-6, TNF-α, and IL-1ß of LPS-induced RAW264.7 cells in a dose-dependent manner. At the same time, the level of the anti-inflammatory cytokine IL-10 was increased after HI-3 treatment. Western blotting results showed that HI-3 suppressed LPS-induced p65 and IκB activation in a dose-dependent manner. Therefore, HI-3 exerts its anti-inflammatory effect by inhibiting the expression of proinflammatory cytokines and the activation of p65 and IκB, which indicated that HI-3 could be a promising therapeutic medicine for inflammation.

The potential of degrading natural chitinous wastes to oligosaccharides by chitinolytic enzymes from two Talaromyces sp. isolated from rotten insects (Hermetia illucens) under solid state fermentation.

It is difficult to produce chitin oligosaccharides by hydrolyzing untreated natural chitinous waste directly. In this study, two fungi Talaromyces allahabadensis Hi-4 and Talaromyces funiculosus Hi-5 from rotten black soldier fly were isolated and identified through multigene phylogenetic and morphological analyses. The chitinolytic enzymes were produced by solid state fermentation, and the growth conditions were optimized by combining single-factor and central composite design. The best carbon sources were powder of molting of mealworms (MMP) and there was no need for additional nitrogen sources in two fungi, then the maximum chitinolytic enzyme production of 46.80 ± 3.30 (Hi-4) and 55.07 ± 2.48 (Hi-5) U/gds were achieved after analyzing the 3D response surface plots. Pure chitin (colloidal chitin) and natural chitinous substrates (represented by MMP) were used to optimize degradation abilities by crude enzymes obtained from the two fungi. The optimum temperature for hydrolyzing MMP (40 °C both in two fungi) were lower and closer to room temperature than colloidal chitin (55 °C for Hi-4 and 45 °C for Hi-5). Then colloidal chitin, MMP and the powder of shrimp shells (SSP) were used for analyzing the products after 5-day degradation. The amounts of chitin oligosaccharides from SSP and MMP were about 1/6 (Hi-4), 1/17 (Hi-5) and 1/8 (Hi-4), 1/10 (Hi-5), respectively, in comparison to colloidal chitin. The main components of the products were GlcNAc for colloidal chitin, (GlcNAc)2 for MMP, and oligosaccharides with higher degree of polymerization (4-6) were obtained when hydrolyzing SSP, which is significant for applications in medicine and health products.

Evaluation of standardized ileal digestibility of amino acids and metabolic availability of methionine, using the indicator amino acid oxidation method, in black soldier fly larvae (Hermetia illucens) meal fed to growing pigs.

Standardized ileal digestibility (SID, %) of crude protein (CP) and amino acid (AA) and the metabolic availability (MA) of Met using the indicator amino acid oxidation (IAAO) method, in partially defatted black soldier fly larvae (PD-BSFL) meal were determined in growing pigs in 2 experiments. The Met SID value was then compared numerically with the Met MA to understand how different SID is compared with its MA value. In Exp. 1, 6 ileal-cannulated barrows (initial body weight [BW] = 18.03 ± 0.34 kg) were used in a 2-period switch back design and fed either a nitrogen-free diet (NFD) or test diet, with PD-BSFL meal as the sole source of AA, over two 11-d experimental periods, at a feeding level of 2.8 × estimated maintenance digestible energy requirement. Barrows were adapted for 9-d to the diet, followed by continuous 8-h ileal digesta collection on day 10 and 11. Digesta were pooled per pig within period. The SID of CP and Met of PD-BSFL meal were 76.1 ± 6.2% and 90.4 ± 3.9%, respectively. In Exp. 2, 7 barrows (initial BW = 18.77 ± 0.69 kg) were used in a 7 × 7 Latin square design with L-[1-13C]-Phe as the indicator AA. Each pig was randomly assigned to 1 of 7 dietary treatments over seven 3-d experimental periods. Two diet types were studied including reference (crystalline AA) and PD-BSFL test diets, each supplying graded intakes of Met at 55, 65, and 75% of the estimated SID requirement (NRC, 2012). The MA of Met was determined by comparing the IAAO response between the reference and PD-BSFL test diet using the slope-ratio method. Linear regression determined a negative slope of the best fit line for both the reference and test diets (P < 0.05). The MA of Met in PD-BSFL meal was 53.3%, which is as expected lower than the SID value. While it is generally appreciated that MA will be less than SID, the use of SID is more practical. In cases where SID cannot explain physiological outcomes of feeding a novel ingredient, IAAO may provide additional insight into whether MA should be explored.

The interest in black soldier fly larvae (BSFL) meal as a protein ingredient in swine feed has grown in the past years. As a novel protein ingredient, it is beneficial to evaluate the amino acid (AA) digestibility and metabolic availability (MA) of the limiting AA, Met, in pigs, in BSFL meal prior to incorporation in feed for a more precise formulation. Two different methodologies were used to determine the AA digestibility and MA, standardized ileal digestibility (SID) and indicator amino acid oxidation (IAAO) method, respectively. Based on the SID values, the AA digestibility of BSFL meal, for some, but not all AA, is comparable to other commonly used protein ingredients in commercial swine feed. When compared with the MA result of Met, the Met SID value is much lower. This indicates that not all digested Met is available for protein synthesis or other metabolic processes in the animal.

Comparison of protein extraction protocols and allergen mapping from black soldier fly Hermetia illucens.

Exploration of important insect proteins - including allergens - and proteomes can be limited by protein extraction buffer selection and the complexity of the proteome. Herein, LC-MS/MS-based proteomics experiments were used to assess the protein extraction efficiencies for a suite of extraction buffers and the effect of ingredient processing on proteome and allergen detection. Discovery proteomics revealed that SDS-based buffer yields the maximum number of protein groups from three types of BSF samples. Bioinformatic analysis revealed that buffer composition and ingredient processing could influence allergen detection. Upon applying multi-level filtering criteria, 33 putative allergens were detected by comparing the detected BSF proteins to sequences from public allergen protein databases. A targeted LC-MRM-MS assay was developed for the pan-allergen tropomyosin and used to assess the influence of buffer composition and ingredient processing using peptide abundance measurements. SIGNIFICANCE: We demonstrated that the selection of protein extraction buffer and the processing method could influence protein yield and cross-reactive allergen detection from processed and un-processed black soldier fly (BSF) samples. In total, 33 putative allergens were detected by comparing the detected BSF proteins to sequences from public allergen protein databases. An LC-MRM-MS assay was developed for tropomyosin, indicating the importance of buffer selection and processing conditions to reduce BSF samples' allergenicity.

Disrupted cellular calcium homeostasis is responsible for Aß-induced learning and memory damage and lifespan shortening in a model of Aß transgenic fly.

Accumulated Aß is one of the hallmarks of Alzheimer's disease. Although accumulated results from in vivo and in vitro studies have shown that accumulated Aß causes learning and memory deficit, cell death, and lifespan reduction, the underlying mechanism remains elusive. In neurons, calcium dynamics is regulated by voltage-gated calcium channel (VGCC) and endoplasmic reticulum and is important for neuron survival and formation of learning and memory. The current study employs in vivo genetics to reveal the role of calcium regulation systems in Aß-induced behavioral damage. Our data shows that although increased VGCC improves learning and memory in Aß42 flies, reduction of VGCC and Inositol trisphosphate receptors extends Aß42 flies' lifespan and improves cell viability. The complex role of calcium regulation systems in Aß-induced damage suggests that the imbalance of calcium dynamic is one of the main factors to trigger learning and memory deficit and cell death in the disease.

Is there any biological insight (or respite) for insects exposed to plastics? Measuring the impact on an insects central carbon metabolism when exposed to a plastic feed substrate.

Insects used to treat organic waste streams and produce valuable protein products are increasingly exposed to plastic contaminated source material assimilating plastic carbon into organic biomass, which is pervasive and hazardous to organisms. Our understanding of this increased insect-plastic interaction remains limited and needs urgent scientific attention if plastic biodegradation and production rates of quality protein are to be improved. Herein, we investigated the biochemical impact of various plastics using three insect models. Black Soldier Fly (BSF), Mealworm (MW), and Wax Moth (WM) larva were each exposed to a plastic substrate (PET, PE, PS, Expanded PE, PP, and PLA) as the primary carbon source for five days to explore any positive metabolic benefits in terms of insect performance and plastic degradation potential. Central carbon metabolism (CCM) metabolites were analyzed via a targeted tMRM liquid chromatography triple quadrupole mass spectrometry (LC-QqQ-MS) method. Unique expressed pathways were observed for each insect model. When reared on PET, BSF larvae were found to have an elevated pyrimidine metabolism, while the purine metabolism pathway was strongly expressed on other plastics. BSF also exhibited a downregulated Vitamin B6 metabolism across all plastics, indicating a likely gut-symbiont breakdown. The MW and WM model insects were metabolically more active on PLA and expanded foam plastics. Further, WM exhibited an elevation in Vitamin B6 metabolism. This data suggests a positive insect-specific interaction towards certain plastic types that warrants further investigation. It is anticipated that through deeper insight into the metabolic impact and benefits afforded from certain plastics, an insect biotransformation pipeline can be established that links fit-for-purpose insect models to individual plastic types that address our growing plastic waste issue.

Indoxacarb resistance-associated mutation of Liriomyza trifolii in Hainan, China.

Liriomyza trifolii, which has been recently prevalent in China, harms more than 300 plant species, especially cowpea in Hainan. This pest also affects the quality and production of vegetables in winter. Indoxacarb is the first commercial oxadiazine pesticide, which is a new efficient insecticide used to control pests of Diptera, including L. trifolii. The unique mechanism of indoxacarb is that indenyl is transformed into N-demethoxycarbonyl metabolite (DCJW) in insects and acts on inactivated sodium channel; DCJW could then destroy the conduction of nerve impulses, which leads to movement disorders, feeding stoppage, paralysis, and eventually the death of pests. The field population of L. trifolii developed resistance by 769 times higher than the sensitive population in Sanya, Hainan. Results revealed the existence of a mutation (i.e., V1848I) in the sixth transmembrane segment of Domain IV of the sodium channel in the field population. The homozygous resistant genotype frequency for the V1848I mutation was 10-15% among the three field-collected populations. This paper reports for the first time the presence of the kdr mutation V1848I in resistant populations of L. trifolii to indoxacarb. The present study will contribute to the understanding of the evolution of indoxacarb resistance and contribute to the development of resistance management practices for winter vegetables in Hainan.