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Nutritional value of black soldier fly ( Hermetia illucens ) larvae (BSFL) processed by three different methods of treatment was compared. The resulting products were the spray-dried BSFL (SPR), oven-dried BSFL 1 (OVN1) and oven-dried BSFL 2 (OVN2). Proximate chemical composition, and profiles of amino acids, fatty acids, minerals, heavy metals, vitamins and nucleotides were analysed and compared. The tested BSFL meals were considered to have a good profile of essential amino acids (EAAs), with leucine, lysine, valine, and histidine being the dominant EAAs. Their content of saturated fatty acids exceeded that of the unsaturated fatty acids. Vitamins B1, B2, and C were also present in the samples. Minerals such as calcium, potassium, phosphorus, sodium, magnesium, zinc, iron, manganese and copper were found to be in adequate amounts in almost all the samples. Heavy metals in the BSFL meals were mostly below 1g kg -1 . Nucleotides such as inosine monophosphate and uridine monophosphate occurred in all the BSFL meals. Other nucleotides, including guanosine monophosphate, adenosine monophosphate, xanthosine monophosphate, and cytidine monophosphate were detected in either or both of SPR and OVN2. In general, the nutritional value of the BSFL meals tested in the present study was influenced by the method of processing.

Antimicrobial peptides from a wide spectrum of insects possess potent microbicidal properties against microbial-related diseases. In this study, seven new gene fragments of three types of antimicrobial peptides were obtained from Hermetia illucens (L), and were named cecropinZ1, sarcotoxin1, sarcotoxin (2a), sarcotoxin (2b), sarcotoxin3, stomoxynZH1, and stomoxynZH1(a). Among these genes, a 189-basepair gene (stomoxynZH1) was cloned into the pET32a expression vector and expressed in the Escherichia coli as a fusion protein with thioredoxin. Results show that Trx-stomoxynZH1 exhibits diverse inhibitory activity on various pathogens, including Gram-positive bacterium Staphylococcus aureus, Gram-negative bacterium Escherichia coli, fungus Rhizoctonia solani Khün (rice)-10, and fungus Sclerotinia sclerotiorum (Lib.) de Bary-14. The minimum inhibitory concentration of Trx-stomoxynZH1 is higher against Gram-positive bacteria than against Gram-negative bacteria but similar between the fungal strains. These results indicate that H. illucens (L.) could provide a rich source for the discovery of novel antimicrobial peptides. Importantly, stomoxynZH1 displays a potential benefit in controlling antibiotic-resistant pathogens.

Black Soldier Fly (BSF) meal is considered as an alternative, emerging and sustainable ingredient for aquafeed production. However, results on fish physiological responses are still fragmentary and often controversial, while no studies are available on fish behavior in response to these new diets. The present work represents the first comprehensive multidisciplinary study aimed to investigate zebrafish physiological and behavioural responses to BSF-based diets. Five experimental diets characterized by increasing inclusion levels (0, 25, 50, 75 and 100% respect to fish meal) of full fat BSF prepupae meal were tested during a 2-months feeding trial. Prepupae were cultured on coffee silverskin growth substrate enriched with a 10% Schizochytrium sp. to improve insects’ fatty acids profile. The responses of zebrafish were assayed through biometric, histological, gas chromatographic, microbiological, spectroscopic, molecular and behavioural analyses. Results evidenced that BSF-based diets affected fish fatty acid composition, while behavioural tests did not show differences among groups. Specifically, a 50% BSF inclusion level diet represented the best compromise between ingredient sustainability and proper fish growth and welfare. Fish fed with higher BSF inclusions (75 and 100%) showed hepatic steatosis, microbiota modification, higher lipid content, fatty acid modification and higher expression of immune response markers.

Background Antimicrobial resistance is currently a major global issue. As the rate of emergence of antimicrobial resistance has superseded the rate of discovery and introduction of new effective drugs, the medical arsenal now is experiencing shortage of effective drugs to combat diseases, particularly against diseases caused by the dreadful multidrug-resistant strains, such as the methicillin-resistant Staphylococcus aureus (MRSA). The ability of fly larvae to thrive in septic habitats has prompted us to determine the antibacterial activity and minimum inhibitory concentrations (MICs) of larval extract of flies, namely Lucilia cuprina, Sarcophaga peregrina and Musca domestica against 4 pathogenic bacteria [ Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Pseudomonas aeruginosa and Escherichia coli ] via a simple and sensitive antibacterial assay, resazurin-based turbidometric (TB) assay as well as to demonstrate the preliminary chemical profile of larval extracts using gas chromatography-mass spectrophotometry (GC-MS). Results The resazurin-based TB assay demonstrated that the L. cuprina larval extract was inhibitory against all tested bacteria, whilst the larval extract of S. peregrina and M. domestica were only inhibitory against the MRSA, with a MIC of 100 mg ml -1 . Subsequent sub-culture of aliquots revealed that the larval extract of L. cuprina was bactericidal against MRSA whilst the larval extracts of S. peregrina and M. domestica were bacteriostatic against MRSA. The GC-MS analysis had quantitatively identified 20 organic compounds (fatty acids or their derivatives, aromatic acid esters, glycosides and phenol) from the larval extract of L. cuprina ; and 5 fatty acid derivatives with known antimicrobial activities from S. peregrina and M. domestica . Conclusion The resazurin-based turbidometric assay is a simple, reliable and feasible screening assay which evidently demonstrated the antibacterial activity of all fly larval extracts, primarily against the MRSA. The larval extract of L. cuprina exerted a broad spectrum antibacterial activity against all tested bacteria. The present study revealed probable development and use of novel and effective natural disinfectant(s) and antibacterial agent(s) from flies and efforts to screen more fly species for antibacterial activity using resazurin-based TB assay should be undertaken for initial screening for subsequent discovery and isolation of potential novel antimicrobial substances, particularly against the multi-drug resistant strains.

The rising global demand for protein-rich food has intensified interest in alternative and sustainable protein sources. Insects, particularly black soldier fly (BSF) larvae, represent promising substrates due to their high nutritional content and potential for valorization into functional ingredients. This study investigated the impact of pre-hydrolysis treatments on the efficiency of enzymatic hydrolysis using alcalase to enhance protein solubilization and bioactive peptide production. Pre-treatments included organic acids (propionic and acetic acid) and a pressure-thermal method. Results indicated that BSF larvae responded differently to the evaluated pre-treatment strategies. Notably, the pressure-thermal treatment combined with enzymatic hydrolysis increased soluble protein content by approximately 30% and antioxidant activity by approximately 20%, suggesting enhanced release of bioactive peptides. Although organic acid treatments increased protein solubility, they did not improve the degree of hydrolysis or antioxidant activity. These findings highlight the potential of pressure-thermal pre-treatment to improve the efficiency of protein extraction from insect biomass and support the integration of such approaches into food bioprocessing strategies aimed at developing novel, high-value protein ingredients.

The aim of this study is to determine to what extent the addition of chitinase to black soldier fly (BSF) larval meal enriched or not with long-chain PUFA (LC-PUFA) could improve growth, protein digestion processes and gut microbial composition in Nile tilapia. Two different types of BSF meal were produced, in which larvae were reared on substrates formulated with vegetable culture substrate (VGS) or marine fish offal substrate (FOS). The BSF raised on VGS was enriched in α-linolenic acid (ALA), while that raised on FOS was enriched in ALA + EPA + DHA. Six BSF-based diets, enriched or not with chitinase, were formulated and compared with a control diet based on fishmeal and fish oil (FMFO). Two doses (D) of chitinase from Aspergillus niger (2 g and 5 g/kg feed) were added to the BSF larval diets (VGD0 and FOD0) to obtain four additional diets: VGD2, VGD5, FOD2 and FOD5. After 53 d of feeding, results showed that the BSF/FOS-based diets induced feed utilisation, protein efficiency and digestibility, as well as growth comparable to the FMFO control diet, but better than the BSF/VGS-based diets. The supplementation of chitinase to BSF/FOS increased in fish intestine the relative abundance of beneficial microbiota such as those of the Bacillaceae family. The results showed that LC-PUFA-enriched BSF meal associated with chitinase could be used as an effective alternative to fishmeal in order to improve protein digestion processes, beneficial microbiota and ultimately fish growth rate.

Blowfly larvae (Diptera: Calliphoridae) have been suggested to have practical application as a toxicological target in forensic science. These larvae may be of use for gunshot residue analysis where traditional analytical targets such as the liver or entry wounds are absent and may allow an opportunity to identify toxins, drugs and gunshot residue from a corpse. A primary weakness of GSR identification in entomotoxicology is the lack of a standardised methodology for processing the larvae for accurate determination of GSR. In this study, Lucilia sericata (Meigen) larvae were exposed to pork mince that was shot 4 times at close range, the larvae were then sampled 12 hourly. A wash protocol for the larvae was developed and the concentrations of Ga, Ba, and Pb, key GSR markers were determined in Lucilia sericata larvae and the solution used to wash them to identify the effectiveness of the cleaning process. Both the whole larvae and each respective wash solution were analysed using inductively coupled plasma mass spectrometry (ICP-MS). Analysis of the wash solutions revealed that a minimum of two washes were required to remove external contaminants prior to ICP-MS analysis of the whole larvae. This work demonstrates the importance of implementing an effective wash protocol when measuring GSR of forensic interest within larvae, as contaminants on the surface of the larvae could lead to misinterpretation of data. [Display omitted] •A wash protocol for larvae was developed removing elements from gunshot residue.•At least two wash cycles are required to surface decontaminate GSR from larvae.•Key gunshot residue elements Ba, Pb and Ga were effective decontamination markers.

Abstract This study evaluated the performance of the lambari-do-rabo-amarelo Astyanax bimaculatus fed diets containing BSF adult fly meal, replacing fishmeal. Two hundred juveniles were distributed in 20 tanks, with five levels of substitution (0%, 12.5%, 25%, 37.5%, and 50%) for 45 days. The diets were isoproteic and isoenergetic, provided to the animals three times a day. The results showed that the increased substitution impaired the fish's performance, with the highest weight gain in the treatment without substitution (0%). Feed conversion, protein efficiency, and energy efficiency decreased due to the high chitin content of the adult fly. It is concluded that BSF fly meal is not advantageous for the lambari-do-rabo-amarelo. Resumo Este estudo avaliou o desempenho do lambari-do-rabo-amarelo Astyanax bimaculatus alimentado com dietas contendo farinha de mosca adulta de BSF, substituindo a farinha de peixe. Duzentos juvenis foram distribuídos em 20 tanques, com cinco níveis de substituição (0%, 12.5%, 25%, 37.5% e 50%), por 45 dias. As dietas foram isoproteicas e isoenergéticas, fornecidas aos animais três vezes ao dia. Os resultados mostraram que a substituição aumentada prejudicou o desempenho dos peixes, com o maior ganho de peso no tratamento sem substituição (0%). A conversão alimentar, eficiência proteica e energética diminuíram, devido ao alto teor de quitina da mosca adulta. Conclui-se que a farinha de mosca de BSF não é vantajosa para o lambari-do-rabo-amarelo.

Forensic entomology uses the age of insects, such as blow flies, to determine a minimum post-mortem interval (PMImin). Recent research has focused on using the analysis of specific cuticular hydrocarbons (CHCs) in adult insects and their empty puparia to estimate their age, as it has been shown that their profile changes are consistent with age. The current work is based on the weathering of five CHCs from empty puparia of Calliphora vicina that were stored in soil (field/outdoor) and non-soil (room/indoor conditions) based pupariation media for a total of six months. The experiment was conducted in a controlled environment chamber at a constant temperature of 25 ± 2 °C under constant darkness. Gas chromatography-mass spectrometry (GC-MS) was used to analyze the cuticular hydrocarbons after they were extracted in n-Hexane. n-Pentacosane, n-Hexacosane, n-Heptacosane, n-Octacosane, and n-Nonacosane were the five CHCs investigated. Results showed that CHCs weathered more quickly in the soil than in the non-soil environment. It was also found that the abundance of Heptacosane increased in the samples during the fifth month when stored in a non-soil medium, while the abundances of all five CHCs were not detected after eight weeks onwards in soil pupation medium. [Display omitted] •Blow flies used to determine PMImin; research focuses on CHCs in insects and puparia.•The study investigated weathering of CHCs in Calliphora vicina puparia stored in soil and non-soil environments.•Gas chromatography-mass spectrometry was used to analyze the cuticular hydrocarbons after extraction in hexane.•CHCs weathered faster in soil; Heptacosane increased in non-soil medium after fifth month.•The relative abundance of all five CHCs were not detected after eight weeks onwards in soil pupation medium.

Widespread resistance of Anopheles sp. populations to pyrethroid insecticides has led to the search for sustainable alternatives in the plant kingdom. Among many botanicals, there is great interest in essential oils and their constituents. Many researchers have explored essential oils (EOs) to determine their toxicity and identify repellent molecules that are effective against Anopheles populations. Essential oils are volatile and fragrant substances with an oily consistency typically produced by plants. They contain a variety of volatile molecules such as terpenes and terpenoids, phenol-derived aromatic components and aliphatic components at quite different concentrations with a significant insecticide potential, essentially as ovicidal, larvicidal, adulticidal, repellency, antifeedant, growth and reproduction inhibitors. The current review provides a summary of chemical composition of EOs, their toxicity at different developmental stages (eggs, larvae and adults), their repellent effects against Anopheles populations, for which there is little information available until now. An overview of antagonist and synergistic phenomena between secondary metabolites, the mode of action as well as microencapsulation technologies are also given in this review. Finally, the potential use of EOs as an alternative to current insecticides has been discussed.