Search

The industry of insect-based proteins as feed and food products has been encountering a huge development since the last decade, and industrial-scale factories are now arising worldwide. Among all the species studied, Black Soldier Fly is one of the most promising and farmed. This rearing activity generates several by-products in the form of chitin-rich biomass that can be valorised to keep a virtuous production cycle embedded in the scope of the bioeconomy. Herein, we report the isolation of chitin and, for the first time, chitin nanocrystals (ChNCs) from all the BSF rearing by-products, i.e., moults (larval exuviae, puparium) and dead adults. Extraction yields, were dependent on the type of by-products and ranged from 5.8 % to 20.0 %, and the chemical structure of the extracts exhibited typical features of α-chitin, confirmed by FTIR, NMR, XRD and TGA analysis. Both STEM in SEM and AFM analysis confirmed the isolation of chitin nanocrystals presenting a rod-like morphology. The average nanocrystal height estimated by AFM ranged from 13 to 27 nm depending on the by-product sample. The following results highlighted the potential of BSF rearing by-products, promoting an approach to valorise those industrial waste and paving the way towards insect-based biorefinery., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Susana Fernandes reports financial support was provided by French National Association for Research and Technology. Susana Fernandes reports financial support was provided by French National Association for Research. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Edible insects with high fat and phosphorus content are a potential novel source of lecithin, however, studies on their minor lipids are limited. In this study, lecithin was extracted from black soldier fly larvae and yellow mealworm. Herein, the effects of lecithin extraction method, matrix and ultrasound pretreatment were explored based on the fatty acid composition and phospholipid profile with soy lecithin as a reference. The use of a wet matrix and ultrasound pretreatment increased the extraction efficiency of total PLs from both insects. Insect lecithin contained a considerable amount of sphingomyelin compared to soy lecithin. In insect lecithin, a total of 47 glycerophospholipid and sphingomyelin molecular species, as well as four molecular species of fatty acyl esters of hydroxy fatty acid, were detected. This study is the first comprehensive investigation of insects as a new source of lecithin with applications in food, cosmetics and in the pharmaceutical industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Odorant-binding proteins (OBPs) in insects are key to detection and recognition of external chemical signals associated with survival. OBP7 in Spodoptera frugiperda's larval stage (SfruOBP7) may search for host plants by sensing plant volatiles, which are important sources of pest attractants and repellents. However, the atomic-level basis of binding modes remains elusive., Results: SfruOBP7 structure was constructed through homology modeling, and complex models of six plant volatiles ((E)-2-hexenol, α-pinene, (Z)-3-hexenyl acetate, lauric acid, O-cymene and 1-octanol) and SfruOBP7 were obtained through molecular docking. To study the detailed interactions between the six plant volatile molecules and SfruOBP7, we conducted three 300 ns molecular dynamics simulations for each study object. The correlation coefficients between binding free energy obtained by molecular mechanics/generalized Born surface area together with solvated interaction energy methods and experimental values are 0.90 and 0.88, respectively, showing a good correlation. By comparing binding free energy along with interaction patterns between SfruOBP7 and the six volatile molecules, hotspot residues of SfruOBP7 when binding with different volatile molecules were determined. Hydrophobic interactions stemming from van der Waals interactions play a significant role in SfruOBP7 and these plant volatile systems., Conclusion: The optimized three-dimensional structure of SfruOBP7 and its binding modes with six plant volatiles revealed their interactions, thus providing a means for estimating the binding energies of other plant volatiles. Our study will help to guide the rational design of effective and selective insect attractants. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The growing world population necessitates the implementation of appropriate processing technologies for edible insects. The objective of this study was to examine the impact of distinct drying techniques, including convective drying at 70 °C (70CD) and 90 °C (90CD) and freeze-drying (FD), on the drying kinetics, physical characteristics (water activity, color), chemical characteristics (chemical composition, amino acid profile, oil properties, total polyphenol content and antioxidant activity, mineral composition, FTIR), and presence of hazards (allergens, microorganisms) of blanched yellow mealworm larvae. The freeze-drying process results in greater lightness and reduced moisture content and water activity. The study demonstrated that the freeze-dried insects exhibited lower contents of protein and essential amino acids as compared to the convective-dried insects. The lowest content of total polyphenols was found in the freeze-dried yellow mealworm larvae; however, the highest antioxidant activity was determined for those insects. Although the oil isolated from the freeze-dried insects exhibited the lowest acid and peroxide values, it proved to have the lowest PUFA content and oxidative stability. All the samples met the microbiological criteria for dried insects. The results of the study demonstrate that a high temperature during the CD method does not result in the anticipated undesirable changes. It appears that freeze-drying is not the optimal method for preserving the nutritional value of insects, particularly with regard to the quality of protein and oil.

The quantification of physical properties of biological matter gives rise to novel ways of understanding functional mechanisms. One of the basic biophysical properties is the mass density (MD). It affects the dynamics in sub-cellular compartments and plays a major role in defining the opto-acoustical properties of cells and tissues. As such, the MD can be connected to the refractive index (RI) via the well known Lorentz-Lorenz relation, which takes into account the polarizability of matter. However, computing the MD based on RI measurements poses a challenge, as it requires detailed knowledge of the biochemical composition of the sample. Here we propose a methodology on how to account for assumptions about the biochemical composition of the sample and respective RI measurements. To this aim, we employ the Biot mixing rule of RIs alongside the assumption of volume additivity to find an approximate relation of MD and RI. We use Monte-Carlo simulations and Gaussian propagation of uncertainty to obtain approximate analytical solutions for the respective uncertainties of MD and RI. We validate this approach by applying it to a set of well-characterized complex mixtures given by bovine milk and intralipid emulsion and employ it to estimate the MD of living zebrafish (Danio rerio) larvae trunk tissue. Our results illustrate the importance of implementing this methodology not only for MD estimations but for many other related biophysical problems, such as mechanical measurements using Brillouin microscopy and transient optical coherence elastography., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

In myrmecophilous organisms, which live in symbiosis with ants, cuticular hydrocarbons (CHCs) play a pivotal role in interspecific communication and defense against chemical-oriented predators. Although these interactions form complex information webs, little is known about the influence of biotic environmental factors on the CHC profiles of myrmecophiles. Here, we analyzed the effect of different host plants and tending ants on the larval CHC profile of Synargis calyce (Lepidoptera: Riodinidae), a polyphagous species with facultative myrmecophily. Groups of caterpillars were fed individually with three host plant species (without tending ants), and with two tending ant species. Through gas chromatography analysis, we compared the cuticular profiles of treatments and found a high similarity between plants and caterpillars (65-82%), but a low similarity between caterpillars and their tending ants (30 - 25%). Cluster analysis showed that caterpillars, ants, and plants form distinct groups, indicating that S. calyce caterpillars have their own chemical profile. These results are similar to those observed for Lycaenidae caterpillars indicating that there is functional convergence in the chemical strategies used by myrmecophilous caterpillar species with similar ecology. Also, the results suggest that the cuticular compounds of S. calyce are primarily influenced by their host plants rather than their tending ants. Thus, we propose that these caterpillars present a trade-off between camouflage and directly informing their presence to ants, maintaining their unique chemical profile, though slightly affected by biotic environmental factors., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Insect protein extract is one of the high-quality protein sources and is frequently viewed as a potential nutrition alternative. However, a more precise method for protein measurement is still needed due to protein overestimation by the Kjeldahl method due to the presence of a large amount of chitin in insects. Therefore, we demonstrated the monitoring of chitin and protein extracted from yellow mealworm larvae through the information on molecular vibration obtained using Raman spectroscopy and infrared (IR) spectroscopy. The NH vibration at 3475 cm -1 is the characteristic peak of chitin in defatted product observed in the Raman spectra. The nitrogen-to-protein conversion factor in protein extracted from larvae by the Raman method was determined based on the NH vibration and found to be 5.66 ± 0.01. We also compared these experimental data to theoretical Raman and IR spectra and determined the possible reasons for why nitrogen elements in chitin affect the determination of protein content. The method of sequentially removing fat and protein could provide more accurate quantification of protein and chitin. Raman spectroscopy is feasible for various types of insects with high chitin content. Compared with the Kjeldahl method, the Raman method is a faster and more accurate measurement method. Moreover, it provides the content of impurities, purity, and structural information., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024. Published by Elsevier B.V.)

With the global population on the rise, an escalating interest exists in environmentally sustainable and friendly protein sources. Insects have emerged as multifaceted resources, viewed not only as potential food items, but also as sources of traditional medicines and proteins. This study utilized response surface methodology (RSM) to ascertain the optimal extraction conditions for proteins from Musca domestica used in toad feeding, denoted as MDPs-T. The yield of MDPs-T was elevated to 18.3% ± 0.2% under these optimized conditions. Subsequently, the particle size, ζ-potentials, and structures of MDPs-T were analyzed and compared with the proteins derived from Musca domestica fed on a normal diet (MDPs-ND). This comparative analysis utilized a range of advanced techniques, involving UV spectroscopy, Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), high-performance gel permeation chromatography (HPGPC), and scanning electron microscopy (SEM). The outcomes have revealed a marginal disparity in the physical and chemical properties between MDPs-T and MDPs-ND. Derosination led to a reduction in the particle size of the MDPs by 10.98% to 62.81%. MDPs-T exhibited a higher proportion of low-molecular-weight components relative to MDPs-ND. Additionally, in a comparative analysis of amino acids, MDPs-T displayed a greater abundance of essential and total amino acids relative to MDPs-ND. Consequently, MDPs-T holds potential as a valuable food supplement for human consumption or as a nutrient-rich feed supplement for animals.

Dual-sweeping-frequency ultrasound (DSFU) was utilized in the preparation of polypeptides from housefly (Musca domestica) larvae protein (HLP). Results indicated that ultrasonication (20 ± 2/28 ± 2 kHz, 42 W/L, 25 min) significantly increased peptide yield and DPPH scavenging capacity by 8.25 % and 14.83 %, respectively. Solubility, foaming and emulsification properties of polypeptides were improved by 19.89 %, 33.33 % and 38.74 % over the control; along with notable reduction in particle size and increase in zeta potential. Tertiary structural changes of the sonicated hydrolysates were illustrated by UV and fluorescence spectra. FTIR showed that ultrasonication increased α-helix, β-turn, and random coil by 38.23 %, 46.35 % and 16.36 %, respectively, but decreased β-sheet by 48.03 %, indicating partial unfolding in HLP hydrolysate conformation and reduction in intermolecular interactions. The research results demonstrated that dual-sweeping-frequency ultrasonication has a great prospect in industry application for the purpose of improving enzymolysis efficiency and product quality for housefly larvae protein hydrolysates production., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Neuropeptides are involved in many biological processes in insects. However, it is unclear what role neuropeptides play in Spodoptera litura adaptation to phytochemical flavone. In this study, 63 neuropeptide precursors from 48 gene families were identified in S. litura , including two neuropeptide F genes ( NPFs ). NPFs played a positive role in feeding regulation in S. litura because knockdown of NPFs decreased larval diet intake. S. litura larvae reduced flavone intake by downregulating NPFs . Conversely, the flavone intake was increased if the larvae were treated with NPF mature peptides. The NPF receptor (NPFR) was susceptible to the fluctuation of NPFs. NPFR mediated NPF signaling by interacting with NPFs to regulate the larval diet intake. In conclusion, this study suggested that NPF signaling regulated diet intake to promote S. litura adaptation to flavone, which contributed to understanding insect adaptation mechanisms to host plants and provide more potential pesticidal targets for pest control.

Ex vivo physiological experiments using small insect models such as Drosophila larvae have become increasingly useful to address fundamental biological questions. To perform such experiments, various artificial saline solutions have been developed, but their osmolality varies significantly from one to the next. Such a variation of osmolality stems, in part, from the difficulty of determining the true value of haemolymph osmolality in Drosophila larvae. Thus, there is a pressing need to refine protocols for collecting and measuring the osmolality of the larval haemolymph. Two major obstacles are thought to impede the accurate analysis of haemolymph collected from small insects: melanin formation and gut-derived contamination. Here, we greatly refined existing haemolymph collection methods, evaluated the purity of the collected haemolymph under melanin-free conditions, and concluded that the true value of haemolymph osmolality is close to 306.0 mOsm kg-1 in Drosophila larvae., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Black soldier fly meal in pet diets is gaining acceptance. This study aimed to assess the use of black soldier fly larvae defatted meal (BSFL) and its impact on blood parameters, biochemical markers, organic antioxidant capacity, skin barrier function and skin and coat quality. A cross-over study involved eight beagle dogs with two periods of 50 days each and a washout period of seven days in between. Two approximately iso-nutritive extruded diets were evaluated, the first containing 29.5% BSFL meal and a control diet containing 26% poultry by-product meal (PBP) as protein source. Skin and coat evaluations and blood collections were conducted before and after each period. Skin barrier function was assessed by measurement of trans epidermal water loss (TEWL) and stratum corneum hydration (SCH) in belly and pinna of the dogs on days 0, 15, 30, and 45 of each period. A trend for higher antioxidant effect significant reduction in serum scavenging capacity was found with PBP for BSFL diet trough malondialdehyde and Vitamin E measurement in dog's serum 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay. When fed PBP diet dogs exhibited reduction in serum cholesterol triglycerides and decreased LDL levels after 50 days, while dogs fed BSFL presented significant reduction in ALT. TEWL was significantly reduced in belly and pinna over time when dogs were fed BSFL, and TEWL in belly was significantly lower in dogs fed BSFL in comparison to PBP. while Increased SCH was also higher for the BSFL group observed in the same along the feeding period in comparison to PBP, indicating improved ability of the dogs to retain water and keep skin moisture. Improvement skin barrier function could be related to fatty acids from BSFL and increased sebaceous lipids in skin. These are responsible for to avoid water loss and improve skin protection against microbial insults. Inclusion of BSFL as protein source did not promote negative changes in blood biochemistry and had minor antioxidant effect in healthy dogs. However, it proved effective in improving skin barrier function, making BSFL a valuable alternative protein source for dogs, particularly those with sensitive skin or allergies manifesting on the skin.

Black soldier fly (BSF) larvae have gained significant attention as ingredients for poultry feed to improve value chain circularity and sustainability. Black soldier fly larvae contain bioactive compounds which can potentially improve broiler health and thereby performance. However, the functionality of bioactive compounds likely depends on how larvae are processed prior to feeding and to which extent larvae products are included in the diet. This may explain the variable results reported in literature on broiler performance and carcass characteristics when feeding them different types of BSF larvae products at different inclusion levels. Therefore, the present research aimed to investigate the effects of different BSF larvae products and inclusion levels in diets on performance and carcass characteristics of slow-growing broilers. The experiment started with 1,728 one-day-old slow-growing male broilers (Hubbard JA757). Nine dietary treatments were used, each replicated eight times. One group of broilers was given a control diet. The following BSF larvae products were investigated: live larvae, a combination of BSF larvae meal and oil mimicking the nutritional composition of the live larvae, and BSF larvae meal and oil separately. All insect products were tested at two inclusion levels. All diet programs were nutritionally comparable (isoenergetic and based on balanced levels of digestible essential amino acids). During the 7-wk trial, several performance parameters and carcass characteristics were measured. The results show that comparable or better broiler performance was achieved with the inclusion of BSF larvae products in the diets compared to the control. Based on the feed conversion ratio (FCR), the unprocessed larvae product and the highest inclusion level led to the most favorable results. Carcass characteristics remained unchanged when BSF larvae products were used in the diets compared to the control group, indicating favorable production output. The BSF larvae products investigated seem suitable feed ingredients for broilers at the current levels tested, generating performance benefits., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Edible insect-derived proteins have attracted considerable attention in the food industry owing to their excellent nutritional and bio-functional activities. Herein, ethanol (20, 40, 60, and 80 %)-treated Tenebrio molitor protein (ETMP) was prepared, and its structural, techno-functional, and antioxidant properties were assessed. As the ethanol concentration increased, the molecular weight of the ETMP decreased, and α-helix content decreased whereas that of β-sheet increased, affecting the secondary structure. Ethanol treatment also resulted in changes in the techno-functional properties of edible insect proteins. ETMP showed significant 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical-scavenging activity (p < 0.05), and its antioxidant activity effectively increased the viability of Vero cells damaged by oxidative stress; 20 % ethanol treatment induced the highest antioxidant activity. In conclusion, our results suggest that appropriate ethanol treatment (20 %) increases the antioxidant activity of edible insect proteins, suggesting their potential in food as an alternative protein resource and functional food with excellent antioxidant activity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

This meta-analysis presents an evaluation of substrate sources and their impact on the growth performance of black soldier fly (BSF) larva. The database, compiled from Google Scholar, PubMed, and Science Direct, focuses on data concerning substrate sources, environmental conditions, and the performance parameters of BSF. Seven types of substrates were analyzed, including Feed Waste, Manure, Fruits, Mix, Animal Source, Fermentation Residue, and Food Waste. The Feed Waste group demonstrated the highest DM content, while the highest CP content was found in the Animal Source group. Higher CP and DM content were found in larva meal from Fermentation Residues and Feed Waste diets, respectively. Higher survival rates were observed in BSF larvae fed on Feed Waste, Fermentation Residues, Food Waste, Fruits, Mix, and Manure substrates compared to Vegetable and Animal Source substrates. Fresh larval weight was lower when Manure was used as a feed substrate than in the Animal Source, Feed Waste, and Vegetable substrates. The prepupal Wet Weight was highest in BSF larvae fed on Animal Source, surpassing those fed on Fermentation Residue, Manure, and Vegetable substrates. Substrate CP content exhibited a positive relationship with fresh larva weight, prepupal wet weight, dry larval weight; larval length, mortality until prepupal, protein conversion, feed conversion ratio, food consumption, substrate reduction rate bioconversion ratio, waste reduction index, and efficiency conversion of digested feed in BSF larva. In conclusion, our findings underline that the source and composition of substrates are correlated to the nutritional composition and conversion efficiency of BSF larva meal., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Innovative solutions are needed to limit environmental effect and optimise resource use as food waste generation rises worldwide. This study investigates the potential of upcycling food waste from fresh markets using Black Soldier Fly (Hermetia illucens) larvae (BSFL) as a sustainable approach. This study explored four fresh market food waste substrates for BSFL bioconversion: discarded fish waste (FI), slaughtered chicken waste (CHI), vegetable waste (VEG), and a 1:1:1 combination of all three (MIX). Soybean curd residue (SCR) was treated as the control substrate. The effects on larval growth, nutritional content, and waste bioconversion rates were examined. The larvae growth rate was strongly impacted by waste type, with BSF-fed CHI and MIX gaining 18.0 and 16.7 mg/d, respectively, followed by BSF-fed with SCR (12.2 mg/d), FI (8.9 mg/d) and VEG (7.6 mg/d). The waste type did not substantially alter BSFL length. The survival rate of the BSFL fed with the food waste studied ranges from 95 to 98.47%, with SCR being the highest. Our findings indicated that BSFL can effectively convert a variety of fresh market food waste into valuable biomass. CHI waste produced the highest larval biomass and bioconversion rate followed by MIX, SCR, FI and VEG. The different waste stream has a major influence on BSFL biomass nutrition. BSFL nutritional composition is independent of the substrate's nutritional content, indicating no direct correlation between substrate and BSFL biomass nutritional composition. SCR waste produced the highest protein content of BSFL (50.49%), followed by VEG (32.61%), MIX (32.57%), FI (31.03%) and CHI (29.06%). SCR waste also produced BSFL biomass with lowest lipid content (26.55%) compared to other waste which resulted into BSFL with lipid levels ranging from 42.92% to 53.72%. BSFL-fed with SCR is the most suitable to be used as an alternative animal's feed based on the protein and lipid levels, while defatting procedure is necessary for the other waste-fed BSFL to render it suitability as animal feed alternatives. Based on bioconversion rate, BSFL growth, and lipid content, the MIX and CHI waste might be viable substrates for future research., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Norhidayah Mohd Taufek reports financial support was provided by Ain Aquaculture Sdn Bhd Norhidayah Mohd Taufek reports a relationship with Ain Aquaculture Sdn Bhd that includes: funding grants., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The proteins were mainly derived from Protaetia brevitarsis larval extracts obtained using two empty intestine methods (traditional static method: TSM or salt immersion stress method: SISM) and extraction solvents (water: W or 50 % water-ethanol: W:E), and the proteins were used as objects to investigate the effect of emptying intestine methods on hypolipidemic peptides. The results revealed that the F-2 fractions of protein hydrolysate had stronger in vitro hypolipidemic activity, with the peptides obtained by SISM possessing a stronger cholesterol micelle solubility inhibition rate, especially in SISM-W:E-P. Moreover, a total of 106 peptides were tentatively identified, among which SISM identified more peptides with an amino acid number < 8. Meanwhile, five novel peptides (YPPFH, YPGFGK, KYPF, SPLPGPR and VPPP) exhibited good hypolipidemic activity in vitro and in vivo, among which YPPFH, VPPP and KYPF had strong inhibitory activities on pancreatic lipase (PL) and cholesteryl esterase (CE), and KYPF, SPLPGPR and VPPP could significantly reduce the TG content in Caenorhabditis elegans. Thus, P. brevitarsis can be developed as a naturally derived hypolipidemic component for the development and application in functional foods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Chitin present in the shell of edible insects is a potential source of chitin, lipids, and proteins, and it exerts various biological activities. Thus far, only a few studies have focused on the use of chitin as a source of high-protein-diet oligosaccharides. The use of insect chitin for the production of high-protein-diet oligosaccharides can lessen the reliance on diet crops. Moreover, although chitin composition in Tenebrio molitor larva, pupa, and adult has been extensively investigated, chitin extraction from T. molitor larval whole body and exuvium has received poor attention. The present study compared the effectiveness of two techniques for extracting high-protein-diet chitin oligosaccharide from an edible insect (T. molitor). Two different extraction sequences of chitin from the larval stage (molitor stage larvae) and adult stage (molitor stage adult) of edible T. molitor were investigated. Two processing steps were employed: (a) deproteinization (DEP) and (b) demineralization (DEM) treatments. Differences in the order, conditions, and period of their application resulted in two different chitin extraction procedures. The viscosity, degree of polymerization, and crystallinity index of the chitin extracted using the two procedures were measured, and its chemical components (chitin, ash, protein, fat, and moisture contents) were determined. T. molitor adults and larvae treated sequentially with DEM-DEP demonstrated the greatest yield of approximately 14.62 % ± 0.15 and 6.096 % ± 0.10 %, respectively. By contrast, when treated sequentially with DEP-DEM, the recorded yields were 10.96 % ± 0.18 and 5.31 % ± 0.38, respectively. Differences in the degree of deacetylation between both methods were observed. Additionally, Fourier transform infrared spectroscopy and X-ray diffractometry of the extracted chitin along with a commercial sample revealed consistent chain conformation, mean hydrogen bonding, and crystallinity index. In this way, residues produced by farmed edible insects can be recovered and used as a novel source of chitin., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This experiment aimed to investigate the effects of partial substitution of crude protein from soybean meal (SBM) with black soldier fly (Hermetia illucens) larvae meal (BSFLM) in juvenile rohu (Labeo rohita) and catla (Catla catla). Four isonitrogenous diets (23% crude protein) were formulated to replace 0% (T0), 40% (T40), 80% (T80) and 100% (T100) crude protein from SBM with BSFLM. Triplicate groups of each species (10 fish per replicate) were fed in an eight week growth experiment. After final sampling (n = 20 fish per dietary group), the remaining fish were exposed to bacterial (Staphylococcus aureus) challenge (0.80 CFU/ml) for 15 days. Rohu fed with BSFLM substituted diets showed significantly higher growth and feed conversion ratio as compared to those in T0. Catla fed with BSFLM substituted diets showed slightly higher growth indices. The growth response of rohu to BSFLM substitution was better than that noted in catla in all groups. The chemical composition, amino acids and fatty acids profile, haematological and biochemical parameters, levels of liver function enzymes measured in T0, T40, T80 and T100 were similar between four dietary groups in both species. However, the maximum value of cholesterol and triglycerides were noted in T100 both in catla and rohu. The values of lauric acid, α-linolenic acid, decosahexanoic acid, n3:n6 fatty acids ratio progressively increased with dietary increase of BSFLM in both species. At end of the growth experiment, the levels of catalase, superoxide dismutase and lysozyme increased linearly with the inclusion of BSFLM in both species while malondialdehyde showed similar values between different groups. However, catalase, and superoxide dismutase increased (T0

Recent studies have investigated the use of Black Soldier Fly (BSF) larvae as a promising biological treatment process for high organic content wastewater (i.a. Leachate from municipal solid waste landfill, food processing effluents), achieving both high treatment efficiency and production of secondary resources from larval biomass (i.a. Proteins and lipids). The present study was aimed at achieving a better understanding of how organic concentration and load might influence treatment performance. Larvae were fed with three artificial wastewaters characterised by same organic substances quality (degree of biodegradability and oxidation of the organic content measured respectively as BOD/COD and TOC/COD ratios) but different organic concentrations. Each type of wastewater was tested at four different loads. Treatment performance was assessed by monitoring both larval growth (in terms of weight variation, mortality and prepupation), and variation of wastewater quality and quantity to determine organic substrate consumption (measured in terms of Total Organic Carbon, TOC). Larval starvation was observed in all tests when TOC concentrations dropped below approx. 1000 mg C/L, which, for the tested wastewater, could be assumed as the limit value for adopting BSF larvae process. Substrate concentration in the feed (mgC/L) influenced larval growth (in terms of maximum wet weight, prepupation and mortality) only when organic load was above 10 mgC/larva: the higher the load, the higher the positive impact of the substrate concentration. On the contrary, the specific substrate consumption rate (v S , mgC/larva/day) appeared not to be influenced by substrate concentration but only by the organic load, with a Michaelis Menten like relationship. Accordingly, substrate load can be assumed as a design parameter for BSF treatment process, while substrate concentration might only influence potential resource recovery from larval biomass., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Currently, as a promising alternative protein source, the interest of edible insect protein has been continuously increased. However, the extraction processing had distinct effects on the physicochemical properties and functionalities of this novel and sustainable protein. In this study, Tenebrio molitor larvae protein (TMLP) was extracted via ultrasound (US)-assisted alkaline extraction. The changes of extraction kinetics, physicochemical characteristics, and functional properties of TMLP as a function of US time (10, 20, 30, 40, 50 min) were investigated. The results showed that 30 min US treatment rendered the maximum protein yield (60.04 %) (P < 0.05). Meanwhile, Peleg's model was considered a suitable model to represent the extraction kinetics of TMLP, with a correlation coefficient of 0.9942. Moreover, the protein secondary structure, particle size, and amino acid profiles of TMLP were changed under the US-assisted alkaline extraction process. Additionally, a significant improvement of the functional properties of TMLP extracted with this method was observed compared to traditional alkaline extraction. In conclusion, the present work suggests that US-assisted alkaline extraction could be considered as a potential method to improve the protein yield, quality profiles, and functional properties of TMLP., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Black soldier fly (BSF) larvae (Hermetia illucens) are voracious feeders that can be reared on food waste streams originating from the food industry and retailers. Because these food waste streams are automatically being unpacked in substantial amounts, they can contain microplastics, potentially jeopardising the larvae's chemical safety when applied as compound feed ingredients. During this study, the dynamics of ingestion and excretion of microplastics by BSF larvae reared on substrates containing different contents (w MP  = 0.00, 0.01, 0.10, 0.50, 1.00, 3.00%) of fluorescent blue-labelled microplastics (median size, Dv(50) = 61.5 µm) were monitored. To correlate the particle size with their uptake, larval mouth opening dimensions were measured during the rearing process. In conclusion, it appeared that ingestion of microplastics by BSF larvae depends on initial particle load, mouth size, and consequently also age. The larvae took up between 131 (w MP  = 0.01%) and 4866 (w MP  = 3.00%) particles leading to bioaccumulation factors (BAF) between 0.12 (w MP  = 3.00%) and 1.07 (w MP  = 0.01%). Larvae also appeared to excrete the microplastics, lowering the BAFs to values between 0.01 (w MP  = 3.00%) and 0.54 (w MP  = 0.01%)., (© 2023. The Author(s).)

Edible insects have been considered as a sustainable and novel protein source to replace animal-derived proteins. The present study aimed to extract Tenebrio molitor larvae proteins (TMP) using ultrasound-assisted alkaline extraction (UAE). Effects of different UAE times (10, 20, 30, 40, and 50 min) on the structural properties and in vitro digestibility of TMP were comparatively investigated with the traditional alkaline extraction method. The results revealed that ultrasonication could effectively alter the secondary/tertiary structures and thermal stability of TMP during UAE. The molecular unfolding and subsequent aggregation of TMP during UAE were mainly attributed to the formation of disulfide bonds and hydrophobic interactions. Moreover, TMP extracted by UAE had higher in vitro digestibility and digestion kinetics than those extracted without ultrasound, and the intermediate UAE time (30 min) was the optimal ultrasound parameter. However, longer UAE times (40 and 50 min) lowered the digestibility of TMP due to severe protein aggregation. The present work provides a potential strategy for the extraction of TMP with higher nutritional values., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Black soldier fly larvae meal (BSFLM) is a relatively new ingredient that has attracted attention for application in human, farm, and companion animal nutrition. However, there is limited data on protein quality (PQ) of BSFLM. This study compared PQ in BSFLM with soybean meal (SBM) and fishmeal (FM) using the classical protein efficiency ratio (PER) chick growth assay model. A total of 240-male day-old Ross × Ross broiler chicks were allocated to 6 test diets for 10 d test. The corn starch-dextrose based diets were: 5 test diets containing 10% crude protein (CP) from casein (as a standard assay diet), SBM, FM, or BSFLM without or with (BSFLM+) additional essential amino acids (EAA) to match the levels of the SBM diet on digestible basis. A sixth diet was nitrogen-free (NFD), to enable calculation of net protein ratio (NPR). The birds had ad libitum access to feed and water, body weight (BW) and feed intake (FI) were recorded at the beginning and end of the experimental feeding period. Body weight gain (BWG), gain to feed ratio (G:F), crude protein intake (CPI), PER, and NPR were calculated. The analyzed CP (as fed) was 10.1, 12.8, 9.5, 11.7, and 14.9% for casein, SBM, FM, BSFLM, and BSFLM+ diets, respectively. Birds fed BSFLM+ had greater BWG (P < 0.01) than birds fed other diets. Feed intake was greater (P < 0.01) for birds fed BSFLM+ and FM diets than birds fed other diets. Birds fed BSFLM+ ate (P < 0.01) more CP than birds fed other diets. Diets had no (P > 0.05) effects on PER and NPR. In terms of ranking, the PER was 2.44, 2.38, 2.34, 2.28, and 2.10 g BWG/g CPI for BSFLM+, SBM, FM, casein, and BSLM, respectively. Corresponding values for NPR were 2.69, 2.78, 2.70, 2.74, and 2.48 g BWG/g CPI, respectively. In conclusion, the protein quality of BSFLM without or with additional EAA was comparable with FM and SBM., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Purpose: In the skin, Lucilia sericata maggot excretions/secretions (ES) accelerate wound healing and limit inflammation. This study aimed to determine whether ES have similar beneficial effects at the ocular surface., Methods: Human corneal epithelial cells (HCEC) were cultured with ES and cell viability was determined by the MTT assay. Additionally, mRNA expression of growth factors, antimicrobial peptides (AMPs) and cytokines was assessed by qPCR. ES ability to modulate TLR-induced IL-6 and IL-8 expression was determined by qPCR and ELISA. ES potential to promote corneal healing was evaluated in vitro by a migration assay in HCEC, and in vivo using a mouse model., Results: ES did not impair HCEC viability up to 25 μg/ml. Among the factors evaluated, only hBD-2 was upregulated (2.5-fold) by 1.5 μg/ml ES after 6 hrs (P = 0.04). In HCEC, ES reduced Poly I:C-induced IL-6 and IL-8 mRNA (P ≤ 0.001) and protein (P ≤ 0.0001) expression. A similar effect was observed with Flagellin (TLR5 agonist) but it was less robust for FSL-1 (TLR2/6 agonist) and Pam3CSK4 (TLR1/2 agonist). The greatest in vitro migration effect was observed with 6.2 μg/ml ES after 44 hrs where gap area compared to vehicle was 53.3 ± 3.7% vs. 72.6 ± 5.4% (P = 0.001). In the mouse model, the maximum healing effect was present with 1.5 μg/ml ES after 12 hrs with a wound area of 19.0 ± 2.7% vs. 60.1 ± 21.6% (P = 0.003) or 77% reduction of the wound area compared to the negative control., Conclusions: ES significantly reduce in vitro TLR-induced production of inflammatory cytokines and promote corneal wound healing., (Copyright © 2022. Published by Elsevier Inc.)

Currently, atherosclerosis control is important to prevent future heart attacks or strokes. Protein-enriched extract (PE) from housefly maggots (Musca domestica) can inhibit the development of atherosclerosis partially through its antioxidant effects. Whether PE exerts other anti-atherosclerosis functions remains unclear. Here, PE was found to simultaneously promote cholesterol metabolism effects in apolipoprotein E knockout (ApoE -/- ) mice. Bile acid synthesis plays a key role in regulating cholesterol homeostasis in atherosclerosis. Whether PE alleviates atherosclerosis by promoting bile acid production and consequent cholesterol consumption was further explored. First, 8-week-old male ApoE -/- mice were recruited and fed on a cholesterol-enriched diet. After 8 weeks, these mice were divided into three groups and received gavage administration of PE, simvastatin, and saline for another 8 weeks. Atherosclerosis severity was then assessed. Real-time quantitative polymerase chain reaction and western blot were employed to determine the expression of hepatic ATP-binding cassette transporter A1 (ABCA1), liver X receptor α (LXRα), and peroxisome proliferator-activated receptor-γ (PPAR-γ). Serum levels of high-density lipoprotein-cholesterol (HDL), low-density lipoprotein-cholesterol (LDL), and total cholesterol (TC) were determined by enzyme-linked immunoassay. Results revealed that PE reversed the formation of atherosclerotic lesion; increased the expression of PPAR-γ, LXRα, and ABCA1; increased the amount of bile flow and total bile acid; reduced the serum level of LDL and TC; and increased the level of HDL. In conclusion, enhancement on bile acid production and consequent cholesterol consumption may partially contribute to the anti-atherosclerotic effects of PE. The reversal of PPARγ-LXRα-ABCA1 signaling pathway may be involved in this process., (© 2022 Wiley Periodicals LLC.)

Research on bioconversion based on insects is intensifying as it addresses the problem of reducing and reusing food and industrial waste. To reach this goal, we need to find more means of pairing waste to insects. With this goal, brewers' spent grains (BSG) - a food waste of the brewing industry - paired with the oleaginous biomass of the thraustochytrid Schizochytrium limacinum cultivated on crude glycerol - a major waste of biodiesel production - were successfully used to grow Hermetia illucens larvae. Combining BSG and S. limacinum in the diet in an attempt to design the lipid profile of H. illucens larvae to contain a higher percentage of omega-3 fatty acids is novel. Insect larvae were grown on three different substrates: i) standard diet for Diptera (SD), ii) BSG, and iii) BSG + 10% S. limacinum biomass. The larvae and substrates were analyzed for fatty acid composition and larval growth was measured until 25% of insects reached the prepupal stage. Our data showed that including omega-3-rich S. limacinum biomass in the BSG substrate promoted an increase in larval weight compared to larvae fed on SD or BSG substrates. Furthermore, it was possible, albeit in a limited way, to incorporate omega-3 fatty acids, principally docosahexaenoic acid (DHA) from BSG + S. limacinum substrate containing 20% of DHA into the larval fat (7% DHA). However, H. illucens with this level of DHA may not be suitable if the aim is to get larvae with high omega-3 lipids to feed carnivorous fish., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Metal exposure impairs respiration, increases metabolic demand, and reduces energy storage/fitness in aquatic species. Respiratory impairment and energy storage was examined in acute selenium-exposed Indian major carps, Catla catla, Labeo rohita and Cirrhinus mrigala fry and were correlated with exposure concentrations. Toxicity effects were determined in a renewal bioassay using 96 h lethal selenium concentrations. Species sensitivity distribution (SSD) was also used to derive predicted no-effect concentrations, toxicity exposure ratios, for selenium exposures to early-life fish stages. Mortality was proportional with increasing concentrations. Oxygen consumption and lipid content compared to moisture and ash and of all protein content in tissues of C. catla and C. mrigala indicates that lowered oxygen consumption is directly predictive of lowered lipid content and selenium-induced hypoxia impacts the energy/nutritional status of the early-life stage of carp. This cross-taxa comparison will have major implications for advancing impact assessment and allow better targeting of species for conservation measures., (Copyright © 2022 Elsevier B.V. All rights reserved.)

According to the concept of circular economy, insects represent good candidates as aquafeed ingredients. Nevertheless, there are some potential chemical risks linked with insect consumption. In this study, we reared the teleost Danio rerio , used as an experimental model, with five experimental diets characterized by increasing levels (0%, 25%, 50%, 75%, and 100%) of full-fat Hermetia illucens (Hi) prepupae, substituting for fish meal (FM) and fish oil (FO). We investigated the presence of potentially toxic elements (PTEs) Cd, Pb, Ni, As, and Hg in larval (20 days), juvenile (2 months), and adult (6 months) fish. Quantitative determinations of Cd, Pb, Ni, and As were made with an atomic absorption spectrometer; the total mercury content was determined by a direct mercury analyzer. The substitution of FM and FO with Hermetia illucens meal led to a reduction in the content of some PTEs, such as Pb, As, and Ni, in fishfeed, leading to concentrations below the legal limit of undesirable substances in animal feed. By increasing the Hi meal dietary content, we observed in the Danio rerio specimens an increase in Cd, Pb, and Ni content and a reduction in As content for all life stages. Moreover, a general increase in the content of Cd, Pb, Hg, and Ni from larvae to juvenile was measured, while the shift of Danio rerio from the juvenile to the adult stage involved a significant increase in the content of Pb, Hg, and Ni. Larvae had a reduced ability to bioaccumulate metal(loid)s compared to juveniles and adults. In conclusion, the content of PTEs in Danio rerio is influenced both by the type of diet administered and by the life stage of the animal itself. This research demonstrates the possibility of using Hi prepupae as an aquafeed ingredient without exposing fish to a chemical risk and, in perspective, allows applying these eco-sustainable diets for the breeding of edible fish species, without endangering human health.

Chitosan was prepared by hydrothermal deacetylation from multi-step protein purification chitin based on Trypoxylus dichotomus, for treating heavy metals and antibiotics. Chitosan with higher deacetylation degree and lower molecular weight were synthesized. The adult chitosan was composed of nanofibers arranged more evenly, showing higher yield, thermal stabilities and antimicrobial properties. The adsorption capacities of Cu 2+ and Fe 3+ were 462 and 270 mg/g, lower than 934 mg/g of Pb 2+ . Levofloxacin and tetracycline hydrochloride adsorption capacity were 26 and 22 mg/g, lower than 67 mg/g of sulfamethoxazole. In addition, compared with single pollutants, the adsorption of sulfamethoxazole and Pb 2+ can increase by 6% and 5% when they act as composite contaminants. The adsorption procedure can be well described by pseudo-second-order kinetics and Langmuir isothermal model, indicating it a homogeneous monolayer chemisorption. Therefore, the Trypoxylus dichotomus source chitosan prepared by hydrothermal deacetylation has potential applications in the adsorption of complex pollutants., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Sarcophaga peregrina (Robineau-Desvoidy, 1830), a synanthropic flesh fly species found in different parts of the world, is of medical and forensic importance. Traditional methods of inferring developmental age rely on the life stage of insects and morphological changes. However, once the larvae reach the pupal and adult stage, morphological changes would become barely visible, so that the classic method would be invalid. Here, we studied the cuticular hydrocarbon profile of S. peregrina of the whole life cycle from larval stage to adult stage by GC-MS. Sixty-three compounds with carbon chain length ranging from 8 to 36 were detected, which could be categorized into four classes: n-alkanes, branched alkanes, alkenes, and unknowns. As developmental increased, branched alkanes dominant, and the content of high-molecular-weight hydrocarbons is variable, especially for 2-methyl C19, DiMethyl C21, docosane (C22), and tricosane (C23). This study shows that the composition of CHC could be used to determine the developmental age of S. peregrina and aid in postmortem interval estimations in forensic science., (© Crown copyright 2021.)

Successful host search by parasitic wasps is often mediated by host-associated chemical cues. The ectoparasitoid Holepyris sylvanidis is known to follow chemical trails released by host larvae of the confused flour beetle, Tribolium confusum, for short-range host location. Although the hexane-extractable trails consist of stable, long-chain cuticular hydrocarbons (CHCs) with low volatility, the kairomonal activity of a trail is lost two days after release. Here, we studied whether this loss of kairomonal activity is due to changes in the chemical trail composition induced by microbial activity. We chemically analyzed trails consisting of hexane extracts of T. confusum larvae after different time intervals past deposition under sterile and non-sterile conditions. GC-MS analyses revealed that the qualitative and quantitative pattern of the long-chain CHCs of larval trails did not significantly change over time, neither under non-sterile nor sterile conditions. Hence, our results show that the loss of kairomonal activity of host trails is not due to microbially induced changes of the CHC pattern of a trail. Interestingly, the kairomonal activity of trails consisting of host larval CHC extracts was recoverable after two days by applying hexane to them. After hexane evaporation, the parasitoids followed the reactivated host trails as they followed freshly laid ones. Cryo-scanning electron microscopy showed that the trails gradually formed filament-shaped microstructures within two days. This self-assemblage of CHCs was reversible by hexane application. Our study suggests that the long-chain CHCs of a host trail slowly undergo solidification by a self-assembling process, which reduces the accessibility of CHCs to the parasitoid's receptors as such that the trail is no longer eliciting trail-following behavior., (© 2021. The Author(s).)

The silkworm (Bombyx mori) is an important economic insect that can be used as food in many countries in Asia. However, silkworms and their metabolites are an important source of allergens, which can induce severe allergic reactions. So far, there are no systematic studies on the potential allergens in silkworm and its metabolites. These studies have important guiding significance for the prevention, diagnosis, and treatment of silkworm allergy. The aim of this study was to identify the potential allergens from larva, pupa, moth, silk, slough and feces of silkworm and analyze the sequence homology of silkworm allergens with other allergens identified in the Allergenonline database. We have found 45 potential allergens in silkworm. The results of the homology comparison suggested that silkworm allergens likely cross-react with those of Dermatophagoides farinae, Aedes aegypti, Tyrophagus putrescentiae, Triticum aestivum and Malassezia furfur., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Ethnopharmacological Relevance: Larvae of the rhinoceros beetle (Allomyrina dichotoma) (ADL) are used ethnopharmacologically to treat gut-related disorders in Korea and China since 1596 and are also approved as a safe novel food with high nutritional value., Aim of the Study: We investigated the protective effects of ADL extract against leaky gut disease using a Drosophila model and sought to elucidate the underlying biological mechanisms., Materials and Methods: We examined the protective effects of ADL extract (2 mg/mL) against the leaky gut disease using a dextran sulfate sodium (DSS)-induced leaky gut Drosophila melanogaster model., Results: We found that oral administration of ADL extracts significantly increase the survival rate of DSS-fed Drosophila. Under conditions of DSS-induced gut damage, ADL extract reduced gut cell apoptosis and gut permeability, resulting in the maintenance of gut tissue homeostasis. Furthermore, we observed that oral administration of ADL extract can induce high levels of E-cadherin gene expression and also restored the original membrane localization of DSS-disrupted E-cadherin contiguous with the armadillo., Conclusion: We concluded that ADL extract plays an important role in maintaining gut homeostasis through the up-regulation of E-cadherin and that it may have a protective effect against leaky gut syndrome., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The excretory/secretory (E/S) products released by infective transitory larvae (xL 3 ) of Haemonchus placei have an important biological function in stimulating immune mechanisms during the invasive process. Our objective was to analyse the modulatory activity of 15 and 70 kDa E/S products from H. placei xL 3 . Both E/S products were collected from xL 3 in vitro cultures at 24 and 72 h. Proteins were confirmed by SDS-PAGE, and the corresponding spots were elicited by gel isoelectrofocusing (IEF) and characterised by mass spectrometry. Additionally, flow cytometry of CD4 + /γδ + T cells and immune gene expression were performed by proliferation assays using each E/S product to stimulate lymphocyte and peripheral blood mononuclear cells (PBMCs) from non-infected calves. The IEF results displayed two spots of 7.0 and 5.7 pI for the 15 and 70 kDa products, respectively. Additionally, 29 and 17 peptides from the 15 and 70 kDa E/S products, respectively, were identified with the hypothetical neurotransmitter and enzymatic functions necessary for larval development. The relative expression displayed upregulation of IL4, 5, 6, 8, 10, 13, IFNγ, and FCεR1A genes (from 2.0- to 17.6-fold, p < 0.05) stimulated by the 15 and 70 kDa proteins, indicating specific genes against haemonchosis. Although the percentage of median florescence intensity (MFI%) of CD4 + /γδ + T cells did not change for both E/S products compared to the negative control and concanavalin-A stimulated cells as the positive control (p > 0.05), the 15-kDa protein reduced the levels of both T cells, and the 70-kDa proteins increased the γδ + cells slightly. Additionally, there was increased PBMCs proliferation by the 70 kDa proteins (p < 0.05), denoting the biological role of other immune cells. The 15 and 70 kDa protein E/S products from H. placei xL 3 showed modulation of the immune response, and although more studies are required, they indicate important functions in the host/parasite interaction., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The problem of a growing resistance of bacteria and other microorganisms to conventional antibiotics gave rise to a search for new potent antimicrobial agents. Insect antimicrobial peptides (AMPs) seem to be promising novel potential anti-infective therapeutics. The dipeptide β-alanyl-tyrosine (β-Ala-Tyr) is one of the endogenous insect toxins exhibiting antibacterial activity against both Gram-negative and Gram-positive bacteria. Prior to testing its other antimicrobial activities, it has to be prepared in a pure form. In this study, we have developed a capillary zone electrophoresis (CZE) method for analysis of β-Ala-Tyr isolated from the extract of the hemolymph of larvae of the fleshfly Neobellieria bullata by reversed-phase high-performance liquid chromatography (RP-HPLC). Based on our previously described correlation between CZE and free-flow zone electrophoresis (FFZE), analytical CZE separation of β-Ala-Tyr and its admixtures have been converted into preparative purification of β-Ala-Tyr by FFZE with preparative capacity of 45.5 mg per hour. The high purity degree of the β-Ala-Tyr obtained by FFZE fractionation was confirmed by its subsequent CZE analysis.

Here, we developed an efficient strategy for the production of lauric acid-enriched monoacylglycerol (MAG) via enzymatic glycerolysis using black soldier fly (Hermetia illucens) larvae (BSFL) oil. The effects of the substrate molar ratio, reaction temperature, type of immobilized lipase, and organic solvent on the MAG content and conversion degree of BSFL oil were optimized. The maximum substrate conversion rate (97.88%) and MAG content (70.84%) were obtained in a tert-butanol system at 50 °C with a glycerol/BSFL oil molar ratio of 4:1 by using immobilized MAS1 lipase as a catalyst. The MAG content in the purified product reached 97.7%, with lauric acid accounting for 50.2%. Improved oxidation stability was observed after glycerolysis. Overall, this study provides a new strategy for the preparation of lauric acid-enriched MAG from BSFL oil., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Aeromonas spp. cause many diseases in aquaculture habitats. Hermetia illucens (Hi) larvae were used as feed-in aquacultures and in eradicating pathogenic fish bacteria. In the present study, we applied consecutive extractions of the same biomass of BSFL fat using the acidic water-methanol solution. The major constituents of the sequential extracts (SEs) were free fatty acids (FFAs), and fatty acids derivatives as identified by gas chromatography spectrometry (GC-MS). Our improved procedure enabled gradual enrichment in the unsaturated fatty acids (USFAs) content in our SEs. The present study aimed to compare the composition and antimicrobial properties of SEs. Among actual fish pathogens, A. hydrophila and A. salmonicida demonstrated multiple drug resistance (MDR) against different recommended standard antibiotics: A. salmonicida was resistant to six, while A. hydrophila was resistant to four antibiotics from ten used in the present study. For the first time, we demonstrated the high dose-dependent antibacterial activity of each SE against Aeromonas spp., especially MDR A. salmonicida . The bacteriostatic and bactericidal (MIC/MBC) activity of SEs was significantly enhanced through the sequential extractions. The third sequential extract (AWME3) possessed the highest activity against Aeromonas spp.: inhibition zone diameters were in the range (21.47 ± 0.14-20.83 ± 0.22 mm) at a concentration of 40 mg/mL, MIC values ranged between 0.09 and 0.38 mg/mL for A. hydrophila and A. salmonicida , respectively. AWME3 MBC values recorded 0.19 and 0.38 mg/mL, while MIC 50 values were 0.065 ± 0.004 and 0.22 ± 0.005 mg/mL against A. hydrophila and A. salmonicida , respectively. Thus, the larvae fat from Hermitia illucens may serve as an excellent reservoir of bioactive molecules with good capacity to eradicate the multidrug-resistant bacteria, having promising potential for practical application in the aquaculture field.

Intestinal fibrosis is induced by excessive myofibroblast proliferation and collagen deposition, which has been regarded as a general pathological feature in inflammatory bowel disease (IBD). Therefore, identifying clinical markers and targets to treat and prevent intestinal fibrosis is urgently needed. The traditional Chinese medicine maggot, commonly known as "wu gu chong", has been shown to reduce oxidative stress and alleviate inflammation in chronic colitis. This study investigated the mechanisms underlying the effects of maggot extract (ME) on inflammation-associated intestinal fibrosis in TGF-β1-stimulated human intestinal fibroblasts (CCD-18Co cells) and dextran sodium sulphate (DSS)-induced chronic colitis murine model. To assess the severity of inflammation and fibrosis, histological and macroscopic evaluation were carried out. The results showed that ME was a significant inhibitor of body weight loss and colon length shortening in mice with chronic colitis. In addition, ME suppressed the intestinal fibrosis by downregulating TGF-β1/SMADs pathway via upregulation of Nrf2 expression at both protein and mRNA levels. ME markedly increased the expression of Nrf2, thus resulting in a higher level of HO-1. After treatment with Nrf2 inhibitor (ML385) or siRNA-Nrf2 for deactivating Nrf2 pathway, the protective effects of ME were abolished both in vitro and in vivo . Moreover, the histopathological results for the major organs of DSS mice treated with ME showed no signs of clinically important abnormalities. Treatment with ME had no effect on the viability of CCD-18Co cells, suggesting its low in vitro cytotoxicity. Furthermore, ME could mediate intestine health by keeping the balance of the gut microbes through the enhancement of beneficial microbes and suppression of pathogenic microbes. In conclusion, this is the first ever report demonstrating that ME ameliorates inflammation-associated intestinal fibrosis by suppressing TGF-β1/SMAD pathway via upregulation of Nrf2 expression. Our findings highlight the potential of Nrf2 as an effective therapeutic target for alleviating intestinal fibrosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Wang, Wang, Wang, Wang, Weng, Zhang, Luo, Lu and Wang.)

Leishmaniasis is a neglected tropical disease that affects millions of people around the world. Larval excretion/secretion (ES) of the larvae of flies of the Calliphoridae family has microbicidal activity against Gram-positive and Gram-negative bacteria, in addition to some species of Leishmania. Our study aimed at assessing the in vitro efficacy of Lucilia cuprina larval ES against the promastigote and amastigote forms of Leishmania amazonensis, elucidating possible microbicidal mechanisms and routes of death involved. Larval ES was able to inhibit the viability of L. amazonensis at all concentrations, induce morphological and ultrastructural changes in the parasite, retraction of the cell body, roughness of the cytoplasmic membrane, leakage of intracellular content, ROS production increase, induction of membrane depolarization and mitochondrial swelling, the formation of cytoplasmic lipid droplets and phosphatidylserine exposure, thus indicating the possibility of apoptosis-like death. To verify the efficacy of larval ES on amastigote forms, we performed a phagocytic assay, measurement of total ROS and NO. Treatment using larval ES reduced the percentage of infection and the number of amastigotes per macrophage of lineage J774A.1 at all concentrations, increasing the production of ROS and TNF-α, thus indicating possible pro-inflammatory immunomodulation and oxidative damage. Therefore, treatment using larval ES is effective at inducing the death of promastigotes and amastigotes of L. amazonensis even at low concentrations., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Honeybee larvae have been recognized as nutrient-rich food in many countries. Although glycogen, a storage form of glucose in animals, is synthesized in honeybee larvae, there is no information on the structure of glycan and its biological activity. In this study, we successfully extracted glycogen from honeybee larvae using hot water extraction and investigated the structure and biological activity of glycan. It was found that the molecular weight of glycogen from honeybee larvae is higher than that of glycogen from bovine liver and oysters. In addition, treatment of RAW264.7 cells with glycogen from honeybee larvae resulted in a much higher production of tumor necrosis factor (TNF)-α and interleukin (IL)-6 than treatment with glycogen from either bovine liver or oysters. These results suggest that the high molecular weight glycogen from honeybee larvae is a functional food ingredient with immunomodulatory activity.

Implementing insects, such as the black soldier fly larvae (BSFL), as animal feed commonly includes the previous removal of substantial amounts of fat. This fat may represent an as yet underutilized energy source for livestock. However, transfer of lauric and myristic acid, prevalent in BSFL fat and undesired in human nutrition, into animal-source foods like eggs may limit its implementation. To quantify this, a laying hen experiment was performed comprising five different diets (10 hens/diet). These were a control diet with soybean oil and meal and a second diet with soybean oil but with partially defatted BSFL meal as protein source. The other three diets were based on different combinations of partially defatted BSFL meal and fat obtained by two different production methods. Lauric acid made up half of the BSFL fat from both origins. Both BSFL fats also contained substantial amounts of myristic and palmitic acid. However, in the insect-based diets, the net transfer from diet to egg yolk was less than 1% for lauric acid, whereas the net transfer for myristic and palmitic acid was about 30% and 100%, respectively. The net transfer did not vary between BSFL originating from production on different larval feeding substrates. The results illustrate that hens are able to metabolize or elongate very large proportions of ingested lauric acid and myristic acid, which are predominant in the BSFL lipids (together accounting for as much as 37 mol%), such that they collectively account for less than 3.5 mol% of egg yolk fatty acids., (© 2021 AOCS.)

The volatile compounds from insects ( Tenebrio molitor and Zophobas morio larvae) roasted at 160, 180, or 200 °C and fed with potato starch or blue corn flour were isolated by solid-phase microextraction (SPME), and identified by gas chromatography-mass spectrometry (GC-MS). In the tested material, 48 volatile compounds were determined. Among them, eight are pyrazines, aroma compounds that are formed in food products during thermal processing due to the Maillard reaction. Eleven of the identified compounds influenced the roast, bread, fat, and burnt aromas that are characteristic for traditional baked dishes (meat, potatoes, bread). Most of them are carbonyl compounds and pyrazines. To confirm the contribution of the most important odorants identified, their odor potential activity values (OAVs) and %OAV were calculated. The highest value was noted for isobuthylpyrazine, responsible for roast aroma (%OAV > 90% for samples roasted at lower temperatures), and 2,5-dimethylpyrazine, responsible for burnt aroma (%OAV > 20% for samples roasted at the highest temperature). According to the study, the type of feed did not significantly affect the results of the sensory analysis of roasted insects. The decisive influence was the roasting temperature. The highest scores were achieved for Tenebrio molitor larvae heat-treated at 160 °C.

Development of new effective antimicrobial drugs is still a big challenge to date due to microbial infection remains an inevitable problem against human health. In this study, fatty acids extract of Lucilia sericata larvae (LFAs) was obtained and evaluated by gas chromatograph-mass spectrometry (GC-MS), and its antibacterial activity against Staphylococcus aureus ( S. aureus ) and Streptococcus pneumoniae ( S. pneumoniae ) was investigated. We found that LFAs exhibited effective antibacterial activity against S. aureus and S. pneumoniae with minimal inhibitory concentrations (MICs) of 125 μg/mL and 100 μg/mL, respectively. The bacterial wall and membrane were the main targets, which was confirmed by fluorescence microscopy, scanning electron microscopy and transmission electron microscopy. Furthermore, a notable anti-biofilm activity against S. aureus and S. pneumoniae was also observed, which was able to both prevent biofilm formation and eradicate mature biofilms of these bacteria. As a promising antibacterial agent, LFAs showed good application prospects in clinical practice.

We studied the temperature dependence of accumulation and elimination of two polychlorinated biphenyls (PCBs; PCB-70 and PCB-126) and a commercial mixture of congeners of polybrominated diphenyl ethers (PBDEs; DE-71™)) in Northern leopard frog (Lithobates pipiens) tadpoles. We reared tadpoles at 18, 23, or 27 °C for 5.3 or up to 13.6 weeks (longer at cooler temperature where development is slower) on diets containing the toxicants, each at several different toxicant concentrations, and compared tissue concentrations as a function of food concentration and rearing temperature. Following > 1 month of accumulation, tissue concentrations of all three toxicants in exposed tadpoles were linearly related to dietary concentrations as expected for first order kinetics, with no significant effect of rearing temperature.We also raised free-swimming L. pipiens tadpoles for 14 days on foods containing either toxicant at 18 or 27 °C during an accumulation phase, and then during depuration (declining toxicant) phase of 14 days we provided food without toxicants and measured the decline of toxicants in tadpole tissue. All the congeners were eliminated faster at warmer rearing temperature, as expected. Using Arrhenius' equation, we calculated that the apparent activation energy for elimination of both PCB congeners by tadpoles was 1.21 eV (95% confidence interval 0.6-1.8 eV). We discuss how this value was within the range of estimates for metabolic reactions generally (range 0.2 - 1.2 eV), which might include metabolic pathways for biotransformation and elimination of PCBs. Furthermore, we discuss how the lack of an effect of rearing temperature on tadpole near-steady-state tissue residue levels suggests that faster elimination at the warmer temperature was balanced by faster uptake, which is plausible considering the similar temperature sensitivities (i.e., activation energies) of all these processes. Although interactions between toxicants and temperature can be complex and likely toxicant-dependent, it is plausible that patterns observed in tadpoles might apply to other aquatic organisms. Published data on depuration in 11 fish species eliminating 8 other organic toxicants indicated that they also had similar apparent activation energy for elimination (0.82 ± 0.12 eV; 95% confidence interval 0.56 - 1.08 eV), even though none of those studied toxicants were PCBs or PBDEs. Additional research on toxicant-temperature interactions can help improve our ability to predict toxicant bioaccumulation in warming climate scenarios., (Copyright © 2021. Published by Elsevier B.V.)

Fatty acids are important compounds for insects, but the requirements for essential fatty acids may differ between insect species. Most of the fatty acids are acquired through the insect's diet; therefore, supplementing the diet with baker's yeast (Saccharomyces cerevisiae Meyen ex E.C. Hansen), which produces unsaturated fatty acids, was predicted to affect the fatty acid composition of the insect. The tested insect was the black soldier fly (BSF) (Hermetia illucens L.), that is used as a source of protein and fat in feed. Therefore, there is importance for BSF larvae (BSFL) nutritional composition, especially the unsaturated fatty acids content, which is one of the nutritional limitations for mammalian diets. The dominant fatty acids of the tested BSFL were the saturated fatty acids: lauric, myristic, and palmitic acids, as found in other BSF studies. Oleic acid (c18:1) and linoleic acid (C18:2) were the abundant unsaturated fatty acids in the BSFL. The proportion of linoleic acid was higher in the substrate with the supplemental yeast; however, this did not affect its proportion in the larvae. The higher proportion of linoleic acid may have been exploited as a source for production of saturated lauric acid. Therefore, providing unsaturated fatty acids to the substrate through supplemental baker's yeast is not the most efficient way to increase the proportion of unsaturated fatty acids in the larvae., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America.)

Age determination of insects collected from vertebrate remains is an essential step in estimating time since colonization as related to the post-mortem interval. Long-established methods for making such estimates rely on determining age related to stage of development at the time of collection in relation to conditions experienced. However, such estimates are based on the completion of a stage of development. Methods allowing for more precise estimates of age (i.e., within a stage of development) are sorely needed. This study examined the potential of volatile organic compounds emitted by blow fly, Ch. rufifacies (Macquart), immatures to determine stage of development, which could potentially be used to estimate the age. Volatile organic compounds (VOCs) from the larval and pupal stages of Ch. rufifacies were collected by headspace solid-phase micro-extraction followed by gas chromatography-mass spectrometry (GC-MS). Analyses indicated 37 compounds shift quantitatively, as well as qualitatively, as the larvae and pupae age. Furthermore, compounds, such as 2-ethyl-1-hexanol, phenol, butanoic acid, hexadecanoic acid, octadecanoic acid, 2-methyl propanamide, and 2-methyl butanoic acid, serve as indicator compounds of specific stages within Ch. rufifacies development. This information could be important to determine if these compounds can be used in the field to predict the presence of certain developmental stages, in order to determine the potential of using volatile markers to estimate time of colonization.

Zootherapy is a traditional secular practice among the Guarani-Kaiowá indigenous ethnic group living in Mato Grosso do Sul, Brazil. My people use the oil extracted from larvae of the snout beetle Rhynchophorus palmarum (Linnaeus, 1758) to treat and heal skin wounds and respiratory diseases. Based on this ethnopharmacological knowledge, the chemical composition and antioxidant, antimicrobial, and healing properties of R. palmarum larvae oil (RPLO) were investigated, as well as possible toxic effects, through in vitro and in vivo assays. The chemical composition of the RPLO was determined using gas chromatography coupled with mass spectrometry. The antioxidant activity of RPLO was investigated through the direct 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, and the antimicrobial activity was evaluated against Gram-positive and Gram-negative bacteria that are pathogenic to humans. The healing properties of RPLO were investigated by performing a cell migration assay using human lung fibroblasts (MRC-5), and the toxicity was analyzed, in vivo, using a Caenorhabditis elegans model and MRC-5 cells, in vitro. RPLO contains 52.2% saturated fatty acids and 47.4% unsaturated fatty acids, with palmitic acid (42.7%) and oleic acid (40%) representing its major components, respectively. RPLO possesses direct antioxidant activity, with a half-maximal inhibitory concentration (IC50) of 46.15 mg.ml-1. The antimicrobial activity of RPLO was not observed at a concentration of 1% (v/v). RPLO did not alter the viability of MRC-5 cells and did not exert toxic effects on C. elegans. Furthermore, MRC-5 cells incubated with 0.5% RPLO showed a higher rate of cell migration than that of the control group, supporting its healing properties. Taken together, RPLO possesses direct antioxidant activity and the potential to aid in the healing process and is not toxic toward in vitro and in vivo models, corroborating the safe use of the oil in traditional Guarani-Kaiowá medicine., Competing Interests: The authors have declared that no competing interests exist.

Anisakid nematode larvae (NL) in fish products comprise a risk to human health and, if visible, lead to the rejection of these products by consumers. Therefore, great efforts are being made for the identification of these anisakid larvae to estimate the potential consumer health risk as well as to develop effective detection methods in order to prevent the introduction of heavily infected fish products into the market. The tasks of national reference laboratories include the improvement of detection methods and to promote their further development. As a prerequisite for improved detection, it is important to understand the structural properties of anisakid NL and compounds produced during host-parasite interactions. This review provides an overview of the intrinsic properties of anisakid NL and reports the latest detection methods in published literature. First, in order to define the potentially interesting intrinsic properties of anisakid nematodes for their detection, anatomy and compounds involved in host-parasite interactions are summarised. These can be used for various detection approaches, such as in the medical field or for allergen detection in fish products. In addition, fluorescence characteristics and their use as both established and promising candidates for detection methods, especially in the field of optical sensing technologies, are presented. Finally, different detection and identification methods applied by the fish processing industries and by control laboratories are listed. The review intends to highlight trends and provide suggestions for the development of improved detection and identification methods of anisakid NL in fish products., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The expression patterns of some cytokines were compared by RT-qPCR between lambs with and without Taenia hydatigena larvae vesicular concentrate (ThLVC) administration and subsequent infection with Haemonchus contortus. Lambs that received ThLVC prior to infection with H. contortus showed lower (p < 0.03) cumulative FEC (AUC = 18450 ± 3384) than infected lambs who did not receive ThLVC (AUC = 31081 ± 3277). Lambs infected with H. contortus, in general, overexpressed Th1 and Th2 cytokines in abomasal mucosa and abomasal lymph nodes, which seems to indicate a generalized and nonpolarized activation of the immune response by H. contortus. The main immunomodulatory effects of ThLVC were observed in the abomasal fundic region. The lambs that were given ThLVC prior to infection strongly overexpressed most of the studied cytokines representing the Th1 (IFNγ and IL2) and Th2 profiles (IL4, IL5, IL6 and IL10), proinflammatory cytokines (SOD1 and PRDX6) and IgE receptor; in contrast, lambs that were infected but did not receive ThLVC only moderately overexpressed IFNγ, IL4 and IL6. The absence of the significant overexpression of cytokines in lambs that only received ThLVC suggests that this derived from T. hydatigena does not have a stimulating effect per se; however, the presence of H. contortus did produce the highest expression (p < 0.01) cytokine profile among lambs that received ThLVC prior to infection compared to those who did not receive it, so its effect seems to be immunomodulatory and not only immunostimulatory., (Copyright © 2021 Elsevier B.V. All rights reserved.)