Effects of different black soldier fly larvae products on slow-growing broiler performance and carcass characteristics.

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.

Identification, characterization and hypolipidemic effect of novel peptides in protein hydrolysate from Protaetia brevitarsis larvae.

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.

A metanalysis to evaluate the effects of substrate sources on the nutritional performance of black soldier fly larvae: implications for sustainable poultry feed.

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.

Preparation of housefly (Musca domestica) larvae protein hydrolysates: Influence of dual-sweeping-frequency ultrasound-assisted enzymatic hydrolysis on yield, antioxidative activity, functional and structural attributes.

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, ß-turn, and random coil by 38.23 %, 46.35 % and 16.36 %, respectively, but decreased ß-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.

Dietary replacement of soybean meal with black soldier fly larvae meal in juvenile Labeo rohita and Catla catla: Effects on growth, nutritional quality, oxidative stress biomarkers and disease resistance.

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

Physicochemical characterization of chitin extracted by different treatment sequences from an edible insect.

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.

Ingestion and excretion dynamics of microplastics by black soldier fly larvae and correlation with mouth opening size.

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 (wMP = 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 (wMP = 0.01%) and 4866 (wMP = 3.00%) particles leading to bioaccumulation factors (BAF) between 0.12 (wMP = 3.00%) and 1.07 (wMP = 0.01%). Larvae also appeared to excrete the microplastics, lowering the BAFs to values between 0.01 (wMP = 3.00%) and 0.54 (wMP = 0.01%).

Treatment of wastewater using Black Soldier Fly larvae: Effect of organic concentration and load.

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 (vS, 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.

Ultrasound-assisted alkaline extraction of protein from Tenebrio molitor larvae: Extraction kinetics, physiochemical, and functional traits.

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.

Mechanisms underlying the effects of ultrasound-assisted alkaline extraction on the structural properties and in vitro digestibility of Tenebrio molitor larvae protein.

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.

Comparative protein quality in black soldier fly larvae meal vs. soybean meal and fish meal using classical protein efficiency ratio (PER) chick growth assay model.

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.

Anti-inflammatory and wound healing potential of medicinal maggot excretions/secretions at the ocular surface.

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.

Protein-enriched extracts from housefly (Musca domestica) maggots alleviates atherosclerosis in apolipoprotein E-deficient mice by promoting bile acid production and consequent cholesterol consumption.

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.

New value from food and industrial wastes - Bioaccumulation of omega-3 fatty acids from an oleaginous microbial biomass paired with a brewery by-product using black soldier fly (Hermetia illucens) larvae.

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.

New Eco-Sustainable Feed in Aquaculture: Influence of Insect-Based Diets on the Content of Potentially Toxic Elements in the Experimental Model Zebrafish (Danio rerio).

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.

Effects of short-term selenium exposure on respiratory activity and proximate body composition of early-life stages of Catla catla, Labeo rohita and Cirrhinus mrigala.

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.

A Study of Cuticular Hydrocarbons of All Life Stages in Sarcophaga peregrina (Diptera: Sarcophagidae).

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.

Adsorption properties of heavy metals and antibiotics by chitosan from larvae and adult Trypoxylus dichotomus.

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 Cu2+ and Fe3+ were 462 and 270 mg/g, lower than 934 mg/g of Pb2+. 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 Pb2+ 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.

[The comprehensive studies of Hermetia illucens larvae protein's biological value].

A high protein content in the insect biomass allows to classify this product as a very promising source of protein, comparable in nutritional and biological value with proteins of animal origin. Despite a long history of safe use, in some countries insects are considered a new type of food which safety must be proven before entering the food market. The long-term Russian experience in novel food's research allows to identify the crucial stages, among which, along with toxicological and allergological tests, the protein's biological value determination takes an important place. The conclusion about the biological value of protein is formed on the basis of integrated use of chemical and biological methods, which gist comes down to the study of the nitrogen balance in the growing organism (biological method) and the calculation of the amino acid score (chemical method). The aim of the research was the comprehensive assessment of Black soldier fly (Hermetia illucens) larvae protein's biological value using chemical and biological methods. Material and methods. Biological studies based on measuring of net protein ratio/net protein utilization were performed on 28 growing (between 25-50 days of life) male Wistar rats, with an initial body weight of ~65.5±1.2 g. Rats in the control group (n=14) received a semi-synthetic casein diet with a protein content of ~12% in calories, the test group (n=14) received a diet including an equivalent amount of H. illucens protein. The diet's ingredients were replaced with the consideration of the proteins, fats, and carbohydrates content in the included product following the principle of isocaloricity and isonitrogenicity (by mass fraction of total nitrogen). H. illucens biomass and casein in the test and control groups, respectively, were the main significant sources of nitrogen in the diet. Body weight, feed intake, and fecal and urine nitrogen losses were measured during the experiment. The biological value and digestibility of protein were judged by coefficients of protein efficiency ratio, net protein ratio, true protein digestibility, true protein biological value, true net protein utilization. Chemical studies included studies of the amino acid composition of H. illucens biomass protein and calculation of the digestible indispensable amino acid score (DIAAS). Results. The general condition of animals of the both groups during the whole experiment was satisfactory, the weekly body weight increase corresponded to the level of growth typical for Wistar rats, intergroup differences were not detected. Despite the fact that in a number of indicators the test group animals differed from the control [there were noted a decrease of the net protein ratio (by 5%, p>0.05), true protein digestibility (by 11%, p<0.05), net protein utilization (by 13%, p<0.05), caused by increased excretion of nitrogen with urine (by 8%, p>0.05) and feces (by 186%, p<0.05), with the same amount of nitrogen intake], the test rats' growth rate and the nitrogen's retention degree indicate a relatively high biological value of insect protein. According to the DIAAS, H. illucens protein is characterized by high content of histidine, threonine, valine, isoleucine and leucine (DIAAS=100 and more), and is also a source of sulfur-containing amino acids - methionine and cysteine (DIAAS=86) and lysine (DIAAS=97). Conclusion. The comprehensive studies of Hermetia illucens larvae protein's biological value demonstrated a high protein content, its balanced amino acid composition and high biological value, which allows to consider Hermetia illucens as a potential source of complete dietary protein.

Application of Capillary and Free-Flow Zone Electrophoresis for Analysis and Purification of Antimicrobial ß-Alanyl-Tyrosine from Hemolymph of Fleshfly Neobellieria bullata.

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 ß-alanyl-tyrosine (ß-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 ß-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 ß-Ala-Tyr and its admixtures have been converted into preparative purification of ß-Ala-Tyr by FFZE with preparative capacity of 45.5 mg per hour. The high purity degree of the ß-Ala-Tyr obtained by FFZE fractionation was confirmed by its subsequent CZE analysis.