Edible insects as a source of biopeptides and their role in immunonutrition.

Many edible insect species are attracting the attention of the food industry and consumers in Western societies due to their high content and quality of protein, and consequently, the potential to be used as a more environmentally friendly dietary source could be beneficial for humans. On the other hand, prevention of inflammatory diseases using nutritional interventions is currently being proposed as a sustainable and cost-effective strategy to improve people's health. In this regard, finding bioactive compounds such as peptides with anti-inflammatory properties from sustainable sources (e.g., edible insects) is one area of particular interest, which might have a relevant role in immunonutrition. This review aims to summarize the recent literature on the discovery of immunomodulatory peptides through in vitro studies from edible insects, as well as to describe cell-based assays aiming to prove their bioactivity. On top of that, in vivo studies (i.e., animal and human), although scarce, have been mentioned in relation to the topic. In addition, the challenges and future perspectives related to edible-insect peptides and their role in immunonutrition are discussed. The amount of literature aiming to demonstrate the potential immunomodulatory activity of edible-insect peptides is scarce but promising. Different approaches have been employed, especially cell assays and animal studies employing insect meal as supplementation in the diet. Insects such as Tenebrio molitor or Gryllodes sigillatus are some of the most studied and have demonstrated to contain bioactive peptides. Further investigations, mostly with humans, are needed in order to clearly state that peptides from edible insects may contribute to the modulation of the immune system.

Dietary and Therapeutic Benefits of Edible Insects: A Global Perspective.

Edible insects are gaining traction worldwide for research and development. This review synthesizes a large and well-established body of research literature on the high nutritional value and variety of pharmacological properties of edible insects. Positive benefits of insect-derived products include immune enhancement; gastrointestinal protection; antitumor, antioxidant, and anti-inflammatory capacities; antibacterial activities; blood lipid and glucose regulation; lowering of blood pressure; and decreased risk of cardiovascular diseases. However, the pharmacological mechanisms of these active components of edible insects in humans have received limited research attention. In addition, we discuss health risks (safety); application prospects; regulations and policies governing their production and consumption with a view to promote innovations, intraglobal trade, and economic development; and suggestions for future directions for further pharmacological functional studies. The aim is to review the current state of knowledge and research trends on edible insects as functional ingredients beneficial to the nutrition and health of humans and animals (livestock, aquatic species, and pets).

Bioactive Peptides Derived from Edible Insects: Effects on Human Health and Possible Applications in Dentistry.

Novel foods, including edible insects, are emerging because of their nutritional characteristics and low environmental impacts and could represent a valid alternative source of food in a more sustainable way. Edible insects have been shown to have beneficial effects on human health. Insect-derived bioactive peptides exert antihypertensive, antioxidant, anti-inflammatory, and antimicrobial properties and have protective effects against common metabolic conditions. In this review, the roles of edible insects in human health are reported, and the possible applications of these peptides in clinical practice are discussed. A special mention is given to the role of antimicrobial peptides and their potential applications in controlling infections in orthodontic procedures. In this context, insects' antimicrobial peptides might represent a potential tool to face the onset of infective endocarditis, with a low chance to develop resistances, and could be manipulated and optimized to replace common antibiotics used in clinical practice so far. Although some safety concerns must be taken into consideration, and the isolation and production of insect-derived proteins are far from easy, edible insects represent an interesting source of peptides, with beneficial effects that may be, in the future, integrated into clinical and orthodontic practice.

Current state of insect proteins: extraction technologies, bioactive peptides and allergenicity of edible insect proteins.

This review aims to provide an updated overview of edible insect proteins and the bioactivity of insect-derived peptides. The essential amino acid content of edible insects is compared with well-known protein sources to demonstrate that edible insects have the potential to cover the protein quality requirements for different groups of the population. Then the current methodologies for insect protein extraction are summarized including a comparison of the protein extraction yield and the final protein content of the resulting products for each method. Furthermore, in order to improve our understanding of insect proteins, their functional properties (such as solubility, foaming capacity, emulsifying, gelation, water holding capacity and oil holding capacity) are discussed. Bioactive peptides can be released according to various enzymatic hydrolysis protocols. In this context, the bioactive properties of insect peptides (antihypertensive, antidiabetic, antioxidant and anti-inflammatory properties) have been discussed. However, the allergens present in insect proteins are still a major concern and an unsolved issue for insect-based product consumption; thus, an analysis of cross reactivity and the different methods available to reduce allergenicity are proposed. Diverse studies of insect protein hydrolysates/peptides have been ultimately promoting the utilization of insect proteins for future perspectives and the emerging processing technologies to enhance the wider utilization of insect proteins for different purposes.

Potentiality Assessment of the Acetylcholinesterase-Inhibitory Activity of Olive Oil with an Additive Edible Insect Powder.

Edible insects (Alphitobius diaperinus Panzer, Gryllus campestris, Tenebrio molitor, Chorthippus biguttulus) are rich in nutrients that potentially inhibit acetylcholinesterase (AChE), but also improve cognition. The aim of this study was to evaluate four varied species of freeze-dried edible insects (purchased from a store); their nutrient composition, including fat, total phenolic compounds, vitamins, and antioxidant properties; and the potential inhibitory effect of AChE. An additional goal was to obtain olive oil with the addition of edible insects. Such oil was characterized by high oxidizing properties and showed high affinity to AChE. The results showed that mealworms and grasshoppers had the highest content of fats (PUFA/SFA) and phenolic compounds. These insects also showed a high content of vitamins, which correlated with the highest affinity for AChE. Therefore, they were added as a functional additive to olive oil. Olive oil with the addition of edible insects showed a higher affinity for AChE and enriched the olive oil with vitamin C and B vitamins.

Termites as human foods-A comprehensive review.

Global food production is anticipated to rise along with the growth of the global population. As a result, creative solutions must be devised to ensure that everyone has access to nutritious, affordable, and safe food. Consequently, including insects in diets has the potential to improve global food and nutrition security. This paper aims to share recent findings by covering edible termites as the main aspect, from their consumption record until consumer acceptance. A total of 53 termite species are reported as edible ones and distributed in 6 biogeographic realms. Generally, termites have a nutrient composition that is suitable for human consumption, and cooked termites are a better dietary choice than their raw counterparts. Besides, increasing customer interest in eating termite-based food can be achieved by making it more palatable and tastier through various cooking processes, that is, boiling, frying, grilling, roasting, smoking, and sun-drying. Moreover, edible termites can also be used as a new source of medication by exhibiting antimicrobial activity. Regarding their advantages, it is strongly encouraged to implement a seminatural rearing system to sustain the supply of edible termites. Overall, this paper makes it evident that termites are an important natural resource for food or medicine. Hence, the long-term objective is to stimulate scientific inquiry into the potential of edible insects as an answer to the problem of global food security.

Bioactive Compounds and Antioxidant Composition of Nut Bars with Addition of Various Edible Insect Flours.

Edible insects represent a new functional source of nutrients that can contribute to solving nutritional deficiency problems. The antioxidant potential and bioactive compounds of nut bars with the addition of three edible insects were evaluated. Acheta domesticus L., Alphitobius diaperinus P. and Tenebrio molitor L. flours were used. A 30% share of insect flour in the bars resulted in significantly greater antioxidant activity (TPC increased from 190.19 for standard bars to 309.45 mg catechin/100 g for bars with 30% addition of cricket flour). Insect flour contributed significantly to an increase in 2,5-dihydrobenzoic acid (from 0.12 for bars with a 15% share of buffalo worm flour to 0.44 mg/100 g in the case of bars with a 30% share of cricket flour) and chlorogenic acid in all bars (from 0.58 for bars with a 15% share of cricket flour to 3.28 mg/100 g for bars with a 30% addition of buffalo worm flour), compared to the standard. The highest content of tocopherols was found in bars with cricket flour, compared to standard bars (43.57 and 24.06 mg/100 g of fat, respectively). The dominant sterol in bars enriched with insect powder was cholesterol. The highest amount of it was found in cricket bars, and the lowest in mealworm bars (64.16 and 21.62 mg/100 g of fat, respectively). The enrichment of nut bars with insect flours raises the levels of valuable phytosterols in the final product. The addition of edible insect flours reduced the perception of most sensory attributes of the bars, compared to the standard bar.

Bioactive Peptide Discovery from Edible Insects for Potential Applications in Human Health and Agriculture.

In the past decade, there has been fast-growing interest among researchers to discover bioactive peptides from edible insects and to evaluate their potential applications in the management of human, livestock, and plant health. This review summarizes current knowledge of insect-derived peptides and their potential role in tackling human health issues and solving agriculture problems by protecting crops and livestock against their pathogens. Numerous bioactive peptides have been identified from edible insect species, including peptides that were enzymatically liberated from insect proteins and endogenous peptides that occur naturally in insects. The peptides exhibited diverse bioactivities, encompassing antioxidant, anti-angiotensin-converting enzyme, anti-dipeptidyl peptidase-IV, anti-glucosidase, anti-lipase, anti-lipoxygenase, anti-cyclooxygenase, anti-obesity, and hepatoprotective activities. Such findings point to their potential contribution to solving human health problems related to inflammation, free radical damage, diabetes, hypertension, and liver damage, among others. Although most of the experiments were performed in vitro, evidence for the in vivo efficacy of some peptides is emerging. Evidence of the protective effects of insect-derived endogenous antimicrobial peptides in combating farm animal and plant pathogens is available. The ability of insect-derived endogenous neuropeptides to protect plants against herbivorous insects has been demonstrated as well. Nevertheless, the potency of peptides identified from insect protein hydrolysates in modulating livestock and plant health remains a knowledge gap to be filled.

Edible Insects an Alternative Nutritional Source of Bioactive Compounds: A Review.

Edible insects have the potential to become one of the major future foods. In fact, they can be considered cheap, highly nutritious, and healthy food sources. International agencies, such as the Food and Agriculture Organization (FAO), have focused their attention on the consumption of edible insects, in particular, regarding their nutritional value and possible biological, toxicological, and allergenic risks, wishing the development of analytical methods to verify the authenticity, quality, and safety of insect-based products. Edible insects are rich in proteins, fats, fiber, vitamins, and minerals but also seem to contain large amounts of polyphenols able to have a key role in specific bioactivities. Therefore, this review is an overview of the potential of edible insects as a source of bioactive compounds, such as polyphenols, that can be a function of diet but also related to insect chemical defense. Currently, insect phenolic compounds have mostly been assayed for their antioxidant bioactivity; however, they also exert other activities, such as anti-inflammatory and anticancer activity, antityrosinase, antigenotoxic, and pancreatic lipase inhibitory activities.

Mycotoxin Metabolism by Edible Insects.

Mycotoxins are a group of toxic secondary metabolites produced in the food chain by fungi through the infection of crops both before and after harvest. Mycotoxins are one of the most important food safety concerns due to their severe poisonous and carcinogenic effects on humans and animals upon ingestion. In the last decade, insects have received wide attention as a highly nutritious, efficient and sustainable source of animal-derived protein and caloric energy for feed and food purposes. Many insects have been used to convert food waste into animal feed. As food waste might contain mycotoxins, research has been conducted on the metabolism and detoxification of mycotoxins by edible insects. The mycotoxins that have been studied include aflatoxins, fumonisins, zearalenone (ZEN), vomitoxin or deoxynivalenol (DON), and ochratoxins (OTAs). Aflatoxin metabolism is proved through the production of hydroxylated metabolites by NADPH-dependent reductases and hydroxylases by different insects. ZEN can be metabolized into α- and ß-zearalenol. Three DON metabolites, 3-, 15-acetyl-DON, and DON-3-glucoside, have been identified in the insect DON metabolites. Unfortunately, the resulting metabolites, involved enzymes, and detoxification mechanisms of OTAs and fumonisins within insects have yet to be identified. Previous studies have been focused on the insect tolerance to mycotoxins and the produced metabolites; further research needs to be conducted to understand the exact enzymes and pathways that are involved.

"Food made with edible insects": Exploring the social representation of entomophagy where it is unfamiliar.

Insects have attracted much attention as a novel food source because of their environmental and nutritional advantages. In Latin America, some traditional groups consume insects; but the urban areas of Brazil resist adopting insects as food, like most Western countries. Thus, this study investigated the social representation of edible insects to understand the barrier behind this avoidance and to identify their segmentation in the Brazilian population. Seven hundred and eighty individuals were interviewed in all the Brazilian regions. The interview was composed of a word association task, a risk perception evaluation and two open-ended questions about the subject. According to the structural approach of social representation, Disgust was the primary association with edible insects. Three segments of consumers were identified using the risk perception of eating insects. The first segment, composed mainly of young men with higher educational levels, had the lowest risk perception, and the most positive attitude about entomophagy, their associations were also positive, such as Acceptance and Sustainability; the second had average risk perception and neutral attitude towards edible insects, with neutral and positive associations (e.g., Flavor and Culture); and the third, formed mostly by older women with lower educational level, presented the highest risk perception and the most negative attitude, and exhibited negative associations (Disgust, and Fear). The insect consumption in Brazil would be favored by using species of crickets, grasshoppers, ants, and their fried and roasted styles of preparations. Also, participants would eat insects mainly because of survival and curiosity.

Occurrence of microplastics in edible aquatic insect Pantala sp. (Odonata: Libellulidae) from rice fields.

BACKGROUND: Microplastic (MP) contamination has been discovered in aquatic systems throughout the world. They are well known as contaminants in aquatic species, but there is a gap in understanding about pathways of MP contamination into humans (i.e., through aquatic animals). The goal of this study is to assess MP contamination in an edible aquatic insect (Pantala sp.) living in rice fields. METHODS: A dragonfly larva, Pantala sp. (Odonata: Libellulidae), was tested for MPs. The study concentrated on three distinct anatomical compartments (whole body, gastrointestinal tract, and body without gastrointestinal tract), each of which was examined separately. For the physical identification and chemical analysis of MPs, a stereomicroscope and a Fourier transformed infrared spectroscope (FT-IR) were used, respectively. RESULTS AND DISCUSSION: The microplastics content was 121 in the whole body, 95 in the gastrointestinal tract, and 66 in the body without the gastrointestinal tract, with an average of 1.34 ± 1.11, 1.06 ± 0.77, and 0.73 ± 0.51 abundance/ individual, respectively. The most common MPs discovered during this study were fragments, followed by fibers and rods. The chemical analysis by FT-IR confirmed three different polymers, including polymethyl methacrylate (PMMA), polyethylene terephthalate (PET), and polypropylene (PP). There was no significant difference in MP abundances among the sample types (Kruskal-Wallis chi-squared = 2.774, df = 2, p = 0.250). The findings suggest that eating an edible aquatic insect (Odonata: Pantala sp.) could be one way for humans to ingest MPs.

Three Amazonian palms as underestimated and little-known sources of nutrients, bioactive compounds and edible insects.

Mauritia flexuosa, Bactris gasipaes, and Oenocarpus bataua are among the main palms in the Amazon used for food and medicinal purposes. The food most commonly derived from these are fruits, oil, and the larvae of the insect Rhynchophorus palmarum reared in their trunks. Palm fruits are used for oil extraction as they are rich in saturated fatty acids, fiber, pro-vitamin A, carotenoids, tocopherols, macro and microelements, and polyphenols. Furthermore, the larvae of R. palmarum are rich in lipids, vitamin E, and proteins. This review analyzes the chemical composition of the fruit and oil of these palm species, as well as the R. palmarum larvae that breed in them. Our aim is to present information that is not widely known in order to demonstrate the potential of these palms as sources of plant-based and animal food with high nutritional and functional values.

Potential health benefits of edible insects.

Animal-based foods have traditionally been viewed as dietary staples because they provide many essential nutrients; however, edible insects have the potential to serve as healthy, sustainable alternatives to these because of their nutrient contents. Edible insects may have superior health benefits due to their high levels of vitamin B12, iron, zinc, fiber, essential amino acids, omega-3 and omega-6 fatty acids, and antioxidants. The addition of edible insects such as crickets to the human diet could offer a myriad of environmental and nutritional benefits including an overall reduction in greenhouse gas emissions, decreased agricultural use of land and water, improved prevention and management of chronic diseases like diabetes, cancer, and cardiovascular disease, and enhanced immune function. Future research should aim to understand the beneficial effects of whole insects or insect isolates in comparison to traditional animal- and plant-based foodstuffs. Ultimately, insects have the potential to be used as meat substitutes or dietary supplements, resulting in human health and environmental benefits. The purpose of this review is to provide additional insight on the nutrient composition of edible insects, their potential use as meat substitutes or dietary supplements, the associated health and wellness benefits, and their potential role in exercise performance.

Trap rebuilding by Myrmeleon brasiliensis larvae (Neuroptera: Myrmeleontidae) in response to flooding: the effect of body size / Reconstrucción de trampas por larvas de Myrmeleon brasiliensis (Neuroptera: Myrmeleontidae) tras las inundaciones: efecto del tamaño corporal

Abstract Introduction: Immature forms of the antlion Myrmeleon brasiliensis (Neuroptera, Myrmeleontidae) build traps in dry sandy soil to capture prey. Objective: The aim of the present study was to investigate how the waterlogging of the soil due to rain affects the trap rebuilding and relocation behavior of M. brasiliensis of different sizes. Methods: The study was conducted between July and December 2019. Larvae M. brasiliensis were observed and collected from a forest reserve in the municipality of Aquidauana in the state of Mato Grosso do Sul, Brazil. Results: In the natural environment, most larvae rebuilt their traps in the same location seven days after the simulation of rain, with a smaller diameter than that observed prior to the simulation of rain. In the laboratory, the movements of M. brasiliensis larvae and rebuilding of the traps after the waterlogging of the soil was affected by body size. Larger larvae moved more and were more likely to rebuild their traps. Conclusions: The saturation of the soil affects the foraging of M. brasiliensis larvae, which are impeded from rebuilding their traps for a period. In situations of long periods of saturated soil, the mortality rate of the larvae is high and rebuilding of the traps occurs after the soil dries out, but with a smaller trap size. These data suggest that changes in the rainfall pattern can affect the population structure of M. brasiliensis larvae, with the selection of larger individuals in situations of more severe rains. In this process, the smaller larvae are more affected, as their foraging is impeded.

Resumen Introducción: Los insectos inmaduros de hormiga león de la especie Myrmeleon brasiliensis (Neuroptera, Myrmeleontidae), construyen trampas en el suelo arenoso seco para capturar a sus presas. Objetivo: El objetivo de este trabajo fue analizar cómo la inundación del suelo, como la lluvia, afecta al comportamiento de reconstrucción y de desplazamiento de las trampas de M. brasiliensis según las diferentes tallas. Métodos: El estudio fue conducido entre julio y diciembre del 2019. Las larvas fueron observadas y recolectadas en una Reserva Forestal en la ciudad de Aquidauana, Estado de Mato Grosso del Sur, Brasil. Resultados: Observamos que, en el ambiente natural, siete días después de la lluvia simulada, la mayoría de las larvas reconstruyeron a sus trampas en el mismo sitio y con diámetro más pequeño que el observado antes de la lluvia simulada. En el laboratorio fue observado que después de la inundación del suelo, el desplazamiento de las larvas y la reconstrucción de las trampas es afectado por el tamaño de su cuerpo. Así, larvas de M. brasiliensis más grandes se desplazan más y presentan una probabilidad más grande de reconstruir a sus trampas. Conclusión: De esa manera, la inundación del suelo afecta el forrajeo de las larvas M. brasiliensis, que se quedan impedidas por un tiempo durante la reconstrucción de sus trampas. Además, en situaciones de largos periodos de inundación del suelo, las larvas presentan una alta tasa de mortalidad y la reconstrucción de sus trampas ocurre después que se seca el suelo, pero esas trampas suelen ser más pequeñas. Esos datos sugieren que es posible que variaciones en el patrón de lluvia pueden afectar la estructura poblacional de las larvas M. brasiliensis, pues en situaciones de lluvias más fuertes, las larvas más grandes suelen tener más éxito. En este proceso, las larvas más pequeñas se ven más afectadas, ya que se impide su búsqueda de alimento.

Skin irritation and potential antioxidant, anti-collagenase, and anti-elastase activities of edible insect extracts.

This study aimed to investigate antioxidant, anti-aging, and irritation properties of Thai edible insect extracts, including Bombyx mori, Omphisa fuscidentalis, Euconocephalus sp., Patanga succincta, Acheta domesticus, and Lethocerus indicus. Insects were extracted by 2 different methods, including maceration using ethanol or hexane and digestion using DI water. Then the extracts were determined for protein content using bicinchoninic acid assay and antioxidant activities using 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid), 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, and ferric thiocyanate assays. Anti-aging activities were investigated by determination of collagenase and elastase inhibitory activities using spectrophotometric assay. Maceration by hexane yielded the highest extract content, whereas aqueous extract from digestion possessed the significantly highest protein content and biological activities (p < 0.05). Interestingly, aqueous extracts of A. domesticus possessed the significantly highest biological activities (p < 0.05) with Trolox equivalent antioxidant capacity value of 8.8 ± 0.1 mmol Trolox/mg, DPPH· inhibition of 19.5 ± 3.8%, equivalent concentration of 12.1 ± 0.7 µM FeSO4/mg, lipid peroxidation inhibition of 31.3 ± 2.4%, collagenase inhibition of 60.8 ± 2.1%, elastase inhibition of 17.0 ± 0.1%, and no irritation effect on chorioallantoic membrane and volunteers. Therefore, aqueous extract of A. domesticus would be suggested for further topical product development.

Nutritional composition of larvae of mealworm (Tenebrio molitor L) and crickets (Gryllus assimilis) with potential usage in feed / [Composição nutricional de larvas de Tenebrio molitor e de grilos (Gryllus assimilis) com potencial de uso na alimentação animal]

Atualmente, tem-se discutido a utilização de insetos na alimentação animal devido ao seu potencial para substituir as fontes tradicionais de proteína utilizadas. O objetivo deste trabalho foi determinar a composição nutricional de larvas de Tenebrio molitor e de grilos do gênero Gryllus assimilis. Os teores de energia bruta (kcal/kg), proteína bruta (g/kg), extrato etéreo (g/kg), cinza (g/kg), FDN (g/kg) e FDA (g/kg) encontrados nas larvas de Tenebrio molitor foram de 7.188,6, 490,2, 335,4, 36,8, 71,8, e 64,0 respectivamente; nos grilos (Gryllus assimilis), os valores foram de 5.942,6, 541,3, 75,2, 49,1, 277,8, e 193,0 respectivamente. Os macros e microminerais quantificados foram fósforo, potássio, sódio, cálcio, magnésio, ferro, manganês, zinco e cobre. Nas larvas de Tenebrio molitor, os valores encontrados foram de 8,56 g/kg, 8,39 g/kg, 1,39 g/kg, 0,44 g/kg, 2,3 g/kg, 48,4 mg/kg, 15 mg/kg, 189 mg/kg e 18 mg/kg respectivamente. Para os grilos (Gryllus assimilis), os teores encontrados foram respectivamente de 8,30 g/kg, 11,6 g/kg, 1,10 g/kg, 3,88 g/kg, 0,82 g/kg, 96,8 mg/kg, 23,7 mg/kg, 18,3 mg/kg e 21,7 mg/kg. Larvas de Tenebrio molitor e grilos do gênero Gryllus assimilis podem ser alternativas para reduzir o uso de fontes de proteína vegetal na alimentação animal.(AU)

Assessment of chemical composition and antioxidant properties of defatted flours obtained from several edible insects.

The aims of this study were determined the chemical composition and the antioxidant properties of defatted flours obtained from several commercially available edible insects such as Acheta dosmesticus, Tenebrio molitor, Zophobas morio, and Rhynchophorus ferrugineus to establish their utilization as ingredient in the development of new food products. The proximate composition of flour was determined using AOAC methods while for antioxidant capacity, four different methodologies were employed (DPPH, ABTS, FIC, and FRAP). The total phenolic content and the tannin content were also determined. All flours analyzed had a high protein content with values ranging between 64.17 and 72.55 g/100 g flour. With regard to the antioxidant activity, R. ferrugineus showed the highest values for DPPH, ABTS, and FRAP assays with values of 2.03, 4.93, and 8.46 mg Trolox equivalent/g flour, respectively. For FIC assay, A. dosmesticus and T. molitor had the highest values 0.47 and 0.48 mg EDTA equivalent/g flour. Defatted flours obtained from edible insects analyzed could have several applications as ingredients to the development new foods due to its good nutrient content and as a functional food for the prevention of oxidation.

Gui-A-Gra Attenuates Testicular Dysfunction in Varicocele-Induced Rats via Oxidative Stress, ER Stress and Mitochondrial Apoptosis Pathway.

Gui-A-Gra, a commercial insect powder from Gryllus bimaculatus, is registered as an edible insect by the Korean food and drug administration. The aim of this study was to investigate the effect of Gui-A-Gra on testicular damage induced by experimental left varicocele in male Sprague Dawley rats. A total of 72 rats were randomly divided into the following six groups (12 rats in each group): a normal control group (CTR), a group administrated with Gui-A-Gra 1.63 gm/kg (G1.63), a group administrated with Gui-A-Gra 6.5 gm/kg (G6.5), a varicocele (VC)-induced control group (VC), a VC-induced group administrated with Gui-A-Gra 1.63 gm/kg (VC + G1.63), and a VC-induced group administrated with Gui-A-Gra 6.5 gm/kg (VC + G6.5). Rats were administrated 1.63 or 6.5 gm/kg Gui-A-Gra once daily for 42 days. Indicators of sperm parameters, histopathology, reproductive hormones, oxidative stress, endoplasmic reticulum (ER) stress, inflammation, and mitochondrial apoptosis were analyzed to evaluate effects of Gui-A-Gra on VC-induced testicular dysfunction. Gui-A-Gra administration to VC-induced rats significantly (p < 0.05) increased sperm count and sperm motility, Johnsen score, spermatogenic cell density, serum testosterone, testicular superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase, GPx4, and steroidogenic acute regulatory protein (StAR) level. Moreover, pretreatment with Gui-A-Gra significantly (p < 0.05) decreased terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) positive cells/tubules, serum luteinizing hormone (LH), serum follicle-stimulating hormone (FSH), testicular tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), malondialdehyde (MDA), reactive oxygen species (ROS)/reactive nitrogen species (RNS) level, glucose-regulated protein-78 (Grp-78), phosphorylated c-Jun-N-terminal kinase (p-JNK), phosphorylated inositol-requiring transmembrane kinase/endoribonuclease 1α (p-IRE1α), cleaved caspase-3, and BCL2 associated X protein: B-cell lymphoma 2 (Bax: Bcl2) ratio in VC rats. These results suggest that protective effects of Gui-A-Gra on VC-induced testicular injury might be due to its antioxidant, anti-inflammatory, and androgenic activities that might be mediated via crosstalk of oxidative stress, ER stress, and mitochondrial apoptosis pathway.

Liquid Chromatography-Tandem Mass Spectrometry for the Simultaneous Analysis of 353 Pesticides in the Edible Insect Tenebrio molitor Larvae (Mealworms).

Tenebrio molitor larvae (mealworm) is an edible insect and is considered a future food. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), a novel method for simultaneous analysis of 353 target analytes was developed and validated. Various sample preparation steps including "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) extraction conditions, number of acetonitrile-hexane partitions, and dispersive-solid phase extraction (dSPE) sorbents were compared, and the optimal conditions were determined. In the established method, 5 g of homogenized mealworms was extracted with acetonitrile and treated with QuEChERS EN 15662 salts. The crude extract was subjected to three rounds of acetonitrile-hexane partitioning, and the acetonitrile layer was cleaned with C18 dSPE. The final solution was matrix-matched and injected into LC-MS/MS (2 µL). For target analytes, the limits of quantitation (LOQs) were ≤10 µg/kg, and the correlation coefficient (r2) of calibration was >0.990. In recovery tests, more than 90% of the pesticides showed an excellent recovery range (70-120%) with relative standard deviation (RSD) ≤20%. For more than 94% of pesticides, a negligible matrix effect (within ±20%) was observed. The analytical method was successfully applied and used for the detection of three urea pesticides in 4 of 11 mealworm samples.