Anoplophora graafi longhorn beetle coloration is due to disordered diamond-like packed spheres.

While artificial photonic materials are typically highly ordered, photonic structures in many species of birds and insects do not possess a long-range order. Studying their order-disorder interplay sheds light on the origin of the photonic band gap. Here, we investigated the scale morphology of the Anoplophora graafi longhorn beetle. Combining small-angle X-ray scattering and slice-and-view FIB-SEM tomography with molecular dynamics and optical simulations, we characterised the chitin sphere assemblies within blue and green A. graafi scales. The low volume fraction of spheres and the number of their nearest neighbours are incompatible with any known close-packed sphere morphology. A short-range diamond lattice with long-range disorder best describes the sphere assembly, which will inspire the development of new colloid-based photonic materials.

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.

Intraspecific and interspecific comparison of toxicity of ladybirds (Coleoptera: Coccinellidae) with contrasting colouration.

Ladybirds (Coccinellidae) use toxic compounds, mostly alkaloids in their haemolymph, for defence against predators and other enemies. The toxicity of ladybirds to predators cannot be directly assessed because predators show avoidance reactions without ingesting the beetles. The alkaloid of ladybird Harmonia axyridis showed wide range toxicity to diverse non-target organisms. Thus, we used a quick, inexpensive and easy-to-perform method using bioassays on water flea Daphnia magna for comparative quantification of the toxicity (LD50) of whole body extracts from several species of ladybirds that differ in their warning colouration. Alien invasive aposematic polymorphic ladybird H. axyridis was more toxic than all the other species examined: aposematic Adalia bipunctata > cryptic Cynegetis impunctata > aposematic Coccinella septempunctata > slightly aposematic Calvia quatuordecimguttata. Three month old adults of H. axyridis were 3.8 times more toxic than two week and one month old adults. The two most common colour morphs (non-melanic novemdecimsignata and melanic spectabilis) did not differ in their toxicity. High toxicity of H. axyridis as compared to all other species examined may contribute to the invasiveness of this species.

Insect residues as an alternative and promising source for the extraction of chitin and chitosan.

This work aimed to obtain and characterize chitin and chitosan extracted from the rearing residues of Tenebrio molitor, Zophobas morio, and Blaptica dubia insects in different growth stages in the same rearing cycles chitin and chitosan yielded 11.21 %-20.89 % and 6.26 %-7.07 %, respectively. The deacetylation degrees of chitosan ranged from 75.75 %-89.21 %, and the solubilities from 69.88 %-94.39 %. Infrared spectroscopy corroborated the acquisition of chitin and chitosan and can be used as a semi-quantitative technique for determining the degree of chitosan deacetylation. The X-ray diffraction profiles revealed the presence of α-chitin, and the relative crystalline indices ranged from 65.9 %-89.2 %. Typical TG profiles with two thermal events are observed for chitin and chitosan samples with different residue contents from the extraction procedure. The chitosan solutions exhibited pseudoplastic behavior, with apparent viscosities ranging from 195.96 to 249.86 mPa.s. The characterization results of the biopolymers extracted from insect residues were similar to those obtained from conventional sources. The growth stage influenced the chitin yield and crystallinity index. The results of this study reinforce the feasibility of using alternative sources of chitin and chitosan, providing the use of waste from insect farms and contributing to sustainability and a circular economy.

Novel spirooxindole alkaloid derivatives from the medicinal insect Blaps japanensis and their biological evaluation.

Blapspirooxindoles A-C (1-3), three novel spirooxindole alkaloids with a unique spiro[chromane-4,3'-indoline]-2,2'-dione motif, blapcumaranons A and B (4 and 5), two new 2-cumaranon derivatives, blapoxindoles A-J (6-15), ten new oxindole alkaloid derivatives, along with one known compound (16), were isolated from the whole bodies of Blaps japanensis. Their structures including absolute configurations were determined by using spectroscopic, X-ray crystallographic, and computational methods. Compounds 1-11 and 13 exist as racemic mixtures in nature, and their (-)- and (+)-antipodes were separated by chiral HPLC. Biological evaluations of these compounds were determined with multiple assays including anti-tumor, anti-inflammatory, and renal protection activities in vitro. Several compounds displayed effective activity in one or more assays.

Investigation on Akis granulifera (Coleoptera, Sahlberg, 1823) as a potential source of chitin and chitosan: Extraction, characterization and hydrogel formation.

The majority of studies have focused on the industrial exploitation of marine fisheries waste through the production of natural bioactive bioploymeres such as chitin and chitosan. However, in recent years, beetles are increasingly attracting the interest of scientists as a source of chitin and chitosan for the preparation of hydrogels for sustainable engineering development. In the present work, we focus on the study for the first time a new Moroccan species of beetle (Akis granulifera Sahlberg, 1823), for the extraction of chitin and the elaboration of chitosan. A chemical extraction process was used. Then, physicochemical characterizations by FT-IR, SEM, XRD, 1H NMR, TGA/DSC, Potentiometry, Viscosimetry, and elemental analysis were performed. In addition, to evaluate its physicochemical quality, the elaborated chitosan is combined with alginate to form a hydrogel. This hydrogel was effectively characterized by SEM, DRX and FTIR to show the potential of chitosan from Akis granulifera in biomaterial applications.

A novel approach for the biosynthesis of silver nanoparticles using the defensive gland extracts of the beetle, Luprops tristis Fabricius.

Discovering novel natural resources for the biological synthesis of metal nanoparticles is one of the two key challenges facing by the field of nanoparticle synthesis. The second challenge is a lack of information on the chemical components needed for the biological synthesis and the chemical mechanism involved in the metal nanoparticles synthesis. In the current study, microwave-assisted silver nanoparticle (AgNP) synthesis employing the defensive gland extract of Mupli beetle, Luprops tristis Fabricius (Order: Coleoptera; Family: Tenebrionidae), addresses these two challenges. This study was conducted without killing the experimental insect. Earlier studies in our laboratory showed the presence of the phenolic compounds, 2,3-dimethyl-1,4-benzoquinone, 1,3-dihydroxy-2-methylbenzene, and 2,5-dimethylhydroquinone in the defensive gland extract of L. tristis. The results of the current study show that the phenolic compounds in the defensive gland extract of the beetle has the ability to reduce silver ions into AgNPs and also acts as a good capping and stabilizing agent. A possible mechanism for the reduction of silver nitrate (AgNO3) into AgNPs is suggested. The synthesized AgNPs were characterized by Ultraviolet-Visible (UV-Vis) spectroscopy, Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy energy-dispersive X-ray (SEM-EDX) analysis and high-resolution transmission electron microscopic (HR-TEM) techniques. The stability of biologically synthesized nanoparticles was studied by zeta potential analysis. The TEM analysis confirmed that AgNPs are well dispersed and almost round shaped. The average size of nanoparticle ranges from 10 to 20 nm. EDX analysis showed that silver is the prominent metal present in the nanomaterial solution. The AgNPs synthesized have antibacterial property against both Staphylococcus aureus and Escherichia coli. Radical scavenging (DPPH) assay was used to determine the antioxidant activity of the AgNPs. AgNPs exhibited anticancer activity in a cytotoxicity experiment against Dalton's lymphoma ascites (DLA) cell line.

Five New Cantharidin Derivatives from the Insect Mylabris cichorii L. and Their Potential against Kidney Fibrosis In Vitro.

Five new monoterpenoids including three 1-hydroxymethyl-2-methyl cantharimide-type derivatives (1, 2, and 5) and two 1,2-dimethyl cantharimide-type derivatives (3 and 4), together with three known compounds (6-8) were isolated from the insect Mylabris cichorii Linnaeus. The structures of these new compounds, including their absolute configurations, were characterized by detailed analysis of NMR, chemical derivatization, and quantum chemical ECD calculations. All of the compounds were tested for their biological activity against kidney fibrosis. The results revealed that compounds 2, 4, and 7 could inhibit kidney fibrosis in vitro at 40 µM by inhibiting the expression of fibronectin and collagen I in TGF-ß1-induced NRK-52e cells.

Blapsimidazolium A: A new imidazolium carboxylate derivative with anti-inflammatory activity from medicinal insect Blaps rynchopetera.

Blaps rynchopetera Fairmaire is a medicinal insect of Yi-nationality medicine used for a long time in Yunnan, China. In the present study, a new blapsimidazolium A (1), together with twelve known N-containing compounds (2-13), were isolated from this insect. The structures were elucidated by extensive spectroscopic analyses (1D and 2D NMR, HR-MS) and comparisons with the reported literature. Blapsimidazolium A was identified as racemic mixture by optical rotation and chiral analysis. Blapsimidazolium A (1) has a unique architecture containing an imidazolium carboxylate moiety. The results of molecular docking showed that blapsimidazolium A bound well to IL-1ß, IL-6 and iNOS. The racemates of (±)-blapsimidazolium A (1) exerted anti-inflammatory activity in LPS-stimulated THP-1 cells by significantly decreasing the production of the proinflammatory cytokines IL-1ß, IL-6 and iNOS. This is the first report describing the anti-inflammatory activity of this type imidazolium carboxylate derivative.

Determination of selected elements in two commercially available edible aquatic insects (Coleoptera) and their worldwide updated list.

Many nations and ethnic groups from underdeveloped and developed countries have used insects as human food since ancient times, a practice that has continued into modern times. In many traditionally entomophagous countries, insects are not a "famine food," and demand for edible insects is clearly a "food of choice" rather than a necessity. From a nutritional point of view, depending on the insect species, gender, insect's diet, and seasons, some species of edible insects have significant amounts of fat, protein, carbohydrates, and vitamins, as well as minerals required for human health, thus representing an alternative food source. In addition to their nutritious content, they may also accumulate pesticide residues, microplastics, and heavy metals from their ecosystems. Therefore, this study analyses some mineral (Ca, Fe, Cu, Zn, Se, Co, Cr, As, and Pb) contents of two selected commercialized aquatic edible beetles, Dytiscus marginalis and Cybister tripunctatus (Dytiscidae), via inductively coupled plasma-mass spectrometry (ICP-MS) and reviews the contemporary list of aquatic edible beetles of the world, summarizing the nutritional characteristics of water beetles with a synthesis of the risks and benefits of edible aquatic beetles in recent literature. The mineral composition of the aquatic beetles is briefly discussed. Antinutritional elements (Pb and As) were detected in aquatic insects, but their contents are below toxic levels for humans. A list of 92 edible aquatic beetle species from Dytiscidae, Gyrinidae, Elmidae, Histeridae, Hydrophilidae, Haliplidae, and Noteridae (Coleoptera) was compiled and updated.

Submilligram Level of Beetle Antifreeze Proteins Minimize Cold-Induced Cell Swelling and Promote Cell Survival.

Hypothermic (cold) preservation is a limiting factor for successful cell and tissue transplantation where cell swelling (edema) usually develops, impairing cell function. University of Wisconsin (UW) solution, a standard cold preservation solution, contains effective components to suppress hypothermia-induced cell swelling. Antifreeze proteins (AFPs) found in many cold-adapted organisms can prevent cold injury of the organisms. Here, the effects of a beetle AFP from Dendroides canadensis (DAFP-1) on pancreatic ß-cells preservation were first investigated. As low as 500 µg/mL, DAFP-1 significantly minimized INS-1 cell swelling and subsequent cell death during 4 °C preservation in UW solution for up to three days. However, such significant cytoprotection was not observed by an AFP from Tenebrio molitor (TmAFP), a structural homologue to DAFP-1 but lacking arginine, at the same levels. The cytoprotective effect of DAFP-1 was further validated with the primary ß-cells in the isolated rat pancreatic islets in UW solution. The submilligram level supplement of DAFP-1 to UW solution significantly increased the islet mass recovery after three days of cold preservation followed by rewarming. The protective effects of DAFP-1 in UW solution were discussed at a molecular level. The results indicate the potential of DAFP-1 to enhance cell survival during extended cold preservation.

Z. morio Hemolymph Relieves E. coli-Induced Mastitis by Inhibiting Inflammatory Response and Repairing the Blood-Milk Barrier.

Escherichia coli (E. coli) is a major environmental pathogen causing coliform mastitis, characterized by cell death and mammary tissue damage. Our previous study has shown the antimicrobial effect of Zophobas morio (Z. morio) hemolymph against mastitis pathogens. In this study, we established E. coli-induced cellular and animal models for mastitis, aiming to evaluate the protective effect of Z. morio hemolymph against E. coli-induced mastitis in vivo and in vitro. In mice with E. coli, Z. morio hemolymph attenuated bacterial burden and histopathological impairment, reduced the production of interleukin (IL)-1ß, IL-18, tumor necrosis factor-α (TNF-α) and the ratio of CD4+ T/CD8+ T, and increased the production of IL-2 triggered by E. coli. Z. morio hemolymph also enhanced the integrity of the blood-milk barrier in E. coli-induced mastitis. In E. coli-stimulated porcine mammary epithelial cells, Z. morio hemolymph inhibited E. coli-induced inflammatory responses and upregulated tight junction proteins (ZO-1, Claudin-3 and Occludin). Moreover, we found that the anti-inflammatory effect of Z. morio hemolymph was mediated by inhibiting E. coli-induced NLRP3 inflammasome assembly, Caspase-1 activation, and reversing the inhibitory effect of E. coli on autophagy. Besides, Z. morio hemolymph augmented ATG5/ATG16L1-mediated autophagy activation, negatively regulated NLRP3 inflammasome activation. Our results reveal that Z. morio hemolymph alleviates E. coli-induced mastitis via lessening the inflammatory response by regulating the NLRP3 and ATG5/ATG16L1 signaling pathway, as well as repairing the blood-milk barrier.

Spectrochemistry of Firefly Bioluminescence.

The chemical reactions underlying the emission of light in fireflies and other bioluminescent beetles are some of the most thoroughly studied processes by scientists worldwide. Despite these remarkable efforts, fierce academic arguments continue around even some of the most fundamental aspects of the reaction mechanism behind the beetle bioluminescence. In an attempt to reach a consensus, we made an exhaustive search of the available literature and compiled the key discoveries on the fluorescence and chemiluminescence spectrochemistry of the emitting molecule, the firefly oxyluciferin, and its chemical analogues reported over the past 50+ years. The factors that affect the light emission, including intermolecular interactions, solvent polarity, and electronic effects, were analyzed in the context of both the reaction mechanism and the different colors of light emitted by different luciferases. The collective data points toward a combined emission of multiple coexistent forms of oxyluciferin as the most probable explanation for the variation in color of the emitted light. We also highlight realistic research directions to eventually address some of the remaining questions related to firefly bioluminescence. It is our hope that this extensive compilation of data and detailed analysis will not only consolidate the existing body of knowledge on this important phenomenon but will also aid in reaching a wider consensus on some of the mechanistic details of firefly bioluminescence.

Intrinsic fluorescence from firefly oxyluciferin monoanions isolated in vacuo.

Fireflies, click beetles, and railroad worms glow in the dark. The color varies from green to red among the insects and is associated with an electronically excited oxyluciferin formed catalytically by the luciferase enzyme. The actual color tuning mechanism has been, and still is, up for much debate. One complication is that oxyluciferin can occur in different charge states and isomeric forms. We present here emission spectra of oxyluciferin monoanions in vacuo at both room temperature and at 100 K recorded with a newly developed and unique mass-spectroscopy setup specially designed for gas-phase ion fluorescence spectroscopy. Ions are limited to the phenolate-keto and phenolate-enol forms that account for natural bioluminescence. At 100 K, fluorescence band maxima are at 599 ± 2 nm and 563 ± 2 nm for the keto and enol forms, respectively, and at 300 K about 5 nm further to the red. The bare-ion spectra, free from solvent effects, serve as important references as they reveal whether a protein microenvironment redshifts or blueshifts the emission, and they serve as important benchmarks for nontrivial excited-state calculations.

New Evidence of Canthariphily: Tilloidea transversalis (Coleoptera: Cleridae) Sequestering Cantharidin From Lydus trimaculatus (Coleoptera: Meloidae).

Cantharidin (CTD) is a defensive compound autogenously and exclusively produced by two phylogenetically related beetle families: Meloidae and Oedemeridae. Although this molecule usually acts as a strong deterrent against potential predators and parasites, some arthropod species, collectively named 'canthariphilous species', are attracted to CTD. Some species can sequester CTD from the CTD-producing species, using it as a chemical defense against enemies. The present paper focuses on the first-ever description of canthariphilous interactions between a checkered beetle species (Coleoptera: Cleridae) and a CTD -producing species. Field observations revealed individuals of the phytophagous beetle Tilloidea transversalis (Charpentier, 1825) (Coleoptera: Cleridae) biting individuals of the blister beetle Lydus trimaculatus (Fabricius, 1775) (Coleoptera: Meloidae). Laboratory behavioral experiments followed to verify if this peculiar behavior of T. transversalis also occurs on other co-occurring species. Moreover, chemical analyses were performed to assess whether T. transversalis can sequester CTD. Our results show that T. transversalis only attacks CTD-producing species. However, while chemical analyses prove that T. transversalis can sequester CTD from the hemolymph of L. trimaculatus, some clues (based on a CTD-baited traps sampling) suggest that this beetle, contrarily to other canthariphilous species, does not appear to show a high attraction to pure synthetic CTD. Thus, other unknown signals, alone or in combination with CTD, could be implicated in triggering the canthariphilous behaviors of T. transversalis.

Physicochemical and sensory properties of biscuits formulated with Tenebrio molitor and Alphitobius diaperinus flours.

The use of insects is considered by Food and Agriculture Organization (FAO) as an alternative source of protein for human and animal food in order to reduce the carbon footprint caused by meat production and to supply the expected population increase with food. In this regard, the aim of this study was to analyze the physicochemical aspects (water activity, protein and fat content, size, optical and mechanical properties) of biscuits prepared with two different insect powders (Tenebrio molitor [TM] and Alphitobius diaperinus [AD], in different percentages [0, 13, 17, 20, and 25%]) based on total weight. In addition, the biscuits formulated with the highest insect powder content were subjected to sensory analysis. The results revealed that all the biscuits formulated with insect powders may be labeled as "high in protein" products following Regulation (EC) No. 1924/2006, although they were not as thick as the control biscuits. The L* and b* coordinates were significantly lower when the insect powders were included, which implied a darkening of the biscuits, which were also harder than the control biscuits, regardless the concentration used. According to the sensory analysis, most panelists perceived the biscuits containing insects as too dark and not crunchy enough and an increase in sweetness might improve their acceptability. Finally, further studies should be carried out to analyze the influence of different structural components and flavorings, such as chocolate, in the formulation of biscuits with insect powders to improve their sensory attributes.

Satiety of Edible Insect-Based Food Products as a Component of Body Weight Control.

Among the many aspects determining the nutritional potential of insect-based foods, research into the satiating potential of foods is an important starting point in the design of new functional foods, including those based on edible insects. The aim of this study was to assess the satiating value of products with the addition of freeze-dried insect flour. The test material included wheat pancakes in which corresponding proportions of wheat flour were substituted with 10% Mw, 0% Mw, and 30% Mw of flour from freeze-dried Tenebrio molitor, 10% Bw, 20% Bw, and 30% Bw of flour from Alphitobius diaperinus, and 10% Cr, 20% Cr, and 30% Cr of flour from Acheta domesticus. The study included the characterisation of physico-chemical properties and their effect on the satiating potential of the analysed pancakes. A total of 71 healthy volunteers (n = 39 women, n = 32 men) with no food phobias were qualified for the study. Each subject rated the level of hunger and satiety before and after ingestion at 30 min intervals over the subsequent 180 min on two separate graphical scales. The rating was done on an unstructured 100 mm visual analogue scale (VAS). A portion intended for testing had a value of 240 kcal. The highest average satiety values were noted for the pancakes with an addition of 30% Alphitobius diaperinus (Bw) and with the addition of 20% and 30% addition of Acheta domesticus flour (Cr). The Tenebrio molitor-based products were the least satiating. However, the largest addition of 30% of an insect flour for each variant considerably increased the satiating potential as compared to the control sample. Satiety was influenced the most by the protein content in the test wheat pancakes. The results support the idea of a possible usage of insect-based food products in the composition of obesity treatment diets, carbohydrate-limiting diets, and as alternative sources of protein.

A Symmetrical Diester as the Sex Attractant Pheromone of the North American Click Beetle Parallelostethus attenuatus (Say) (Coleoptera: Elateridae).

Hexanoic acid, 1-octanol, 1,8-octanediol, octyl hexanoate, 1,8-octanediol monohexanoate, and 1,8-octanediol dihexanoate were identified in headspace volatiles collected from the crushed abdomen of a female click beetle of the species Parallelostethus attenuatus (Say) (Elaterinae, tribe Elaterini). In field trials carried out in Illinois, South Carolina, North Carolina, and Virginia, adult male beetles were strongly attracted to 1,8-octanediol dihexanoate alone. Blends of the dihexanoate with one or more of the other compounds proved to be less attractive than the dihexanoate alone, suggesting that the pheromone of this species may consist of a single compound. The symmetrical diester structure of the pheromone is a novel natural product and appears to be structurally unique among insect pheromones.

Pachyrhynchus Weevils Use 3D Photonic Crystals with Varying Degrees of Order to Create Diverse and Brilliant Displays.

The brilliant appearance of Easter Egg weevils, genus Pachyrhynchus (Coleoptera, Curculionidae), originates from complex dielectric nanostructures within their elytral scales and elytra. Previous work, investigating singular members of the Pachyrhynchus showed the presence of either quasi-ordered or ordered 3D photonic crystals based on the single diamond ( Fd3¯m ) symmetry in their scales. However, little is known about the diversity of the structural coloration mechanisms within the family. Here, the optical properties within Pachyrhynchus are investigated by systematically identifying their spectral and structural characteristics. Four principal traits that vary their appearance are identified and the evolutionary history of these traits to identify ecological trends are reconstructed. The results indicate that the coloration mechanisms across the Easter Egg weevils are diverse and highly plastic across closely related species with features appearing at multiple independent times across their phylogeny. This work lays a foundation for a better understanding of the various forms of quasi-ordered and ordered diamond photonic crystal within arthropods.

Pygidial glands of the blue ground beetle Carabus intricatus: chemical composition of the secretion and its antimicrobial activity.

Pygidial gland secretions are used as repellent defensive allomones in ground beetles. We provide the first precise data on the chemical composition and antimicrobial potency of the secretion of the blue ground beetle, as well as on the morphology of its pygidial glands. The latter structures were not previously studied chemoecologically and morphologically, and we hypothesized that their secretion may have some antimicrobial action, as is the case with certain Carabus species. Gas chromatography-mass spectrometry (GC-MS) was used to identify methacrylic and angelic acids as dominant chemicals in the secretion from individuals of three populations of the blue ground beetle in Serbia. We tested its secretion against selected strains of medically important microorganisms. The secretion exibits antimicrobial action against certain bacterial species and all tested micromycetes. The most significant antifungal effect of the secretion was against Penicillium ochrochloron, which is more sensitive to the secretion than to commercial antifungal drugs ketoconazole and bifonazole. Bifonazole achieved minimum inhibitory concentrations against Trichoderma viride at more than three times higher value than did the secretion, indicating a significant antifungal effect of the secretion against this micromycete as well. Additionally, we tested commercially available standards of two dominant chemicals in the secretion to investigate their interaction and antimicrobial role in the secretion. Finally, we describe all glandular morpho-functional units of the blue ground beetle. Our results suggest that the secretion of the blue ground beetle may serve not only defensive but also antimicrobial functions, which likely aid the survival of this beetle in the microbial-rich forest litter habitat.