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In this study, peptides designed using fragments of an antifreeze protein (AFP) from the freeze-tolerant insect Tenebrio molitor, TmAFP, were evaluated as inhibitors of clathrate hydrate formation. It was found that these peptides exhibit inhibitory effects by both direct and indirect mechanisms. The direct mechanism involves the displacement of methane molecules by hydrophobic methyl groups from threonine residues, preventing their diffusion to the hydrate surface. The indirect mechanism is characterized by the formation of cylindrical gas bubbles, the morphology of which reduces the pressure difference at the bubble interface, thereby slowing methane transport. The transfer of methane to the hydrate interface is primarily dominated by gas bubbles in the presence of antifreeze peptides. Spherical bubbles facilitate methane migration and potentially accelerate hydrate formation; conversely, the promotion of a cylindrical bubble morphology by two of the designed systems was found to mitigate this effect, leading to slower methane transport and reduced hydrate growth. These findings provide valuable guidance for the design of effective peptide-based inhibitors of natural-gas hydrate formation with potential applications in the energy and environmental sectors. [ABSTRACT FROM AUTHOR]

A new approach to supplying foods and feeds that are high in protein has emerged in the form of insect farming. Insect‐farmed food may also serve as part of a sustainable diet for humans. The mealworm Tenebrio molitor L. (Coleoptera: Tenebrionidae) is the most widely consumed insect worldwide. In this study, 10 larval diets were tested in the laboratory to determine their suitability for mass‐feeding mealworms at 28 °C and 60–65% r.h., in a completely randomized design. Various larval diets had a significant impact on the biological features of T. molitor. The larvae, pupae, and adults were greater on diets containing wheat bran alone and wheat bran in combination with maize and porridge. Larval mortality was significantly higher (60.7%) on a diet with porridge alone, compared to a control diet, followed by diets with porridge in combination with barley (55.6%) or maize (50.6%). Sex ratio was also affected by the tested larval diets. Female hatch (65%) was higher than male hatch (35%) on diet containing wheat bran alone as compared to all other diets. The findings contribute to optimizing mealworm rearing practices for sustainable food production and promote the potential use of mealworms as a viable protein source – especially livestock diets based on mealworm reared on wheat bran show promise. [ABSTRACT FROM AUTHOR]

Practical Application The use of edible insect protein in food products is contingent on their biological effects. Conventional protein extraction methods are not only time‐consuming and costly but also energy‐intensive. There is a need for alternative techniques that maintain the bioactivities of insect proteins and are environmentally sustainable. This study compares the health functionality of mealworm (Tenebrio molitor larvae) concentrates obtained by conventional methods—alkali and salt (MS) extraction—and nonconventional methods—enzyme (ME) and screw press (MP)—to enhance their applicability despite lower protein concentration. Overall, MP exhibited the highest essential amino acids content, whereas ME showed the highest in vitro digestibility, total phenolic contents, and antioxidant capacities among all the concentrates. ME also had a significant cell proliferative capacity at concentrations ≥500 µg/mL. MS significantly inhibited tumor necrosis factor‐alpha and interleukin‐1beta secretion in lipopolysaccharide‐treated Hep3B cells compared to other samples. As for anti‐hyperglycemia effects, treatment with MS and ME for 2 and 5 min significantly increased the p‐Akt/Akt ratio (MS, 1.34‐ and 1.61‐fold; ME, 2.26‐ and 2.70‐fold, respectively). In conclusion, enzyme treatment enhanced nutritional value and antioxidant capacity, whereas salt treatment potentially contributed to anti‐inflammatory and anti‐hyperglycemia activities. Hybrid extraction techniques combining conventional and nonconventional methods are suggested based on target applications, considering health benefits, environmental impact, costs, and efficiencies.Four mealworm protein extraction methods (alkali/salt/enzyme/screw press) were compared for their nutritional and biological properties. Alkali extraction enhanced protein content, enzyme treatment improved nutritional value and antioxidant capacity, and salt‐assisted extraction exhibited immunomodulatory effects in vitro. Notably, enzyme and salt treatments produced protein concentrates with significant antidiabetic and anti‐hyperglycemic properties. [ABSTRACT FROM AUTHOR]

Iron is an essential mineral that supports biological functions like growth, oxygen transport, cellular function, and hormone synthesis. Insufficient dietary iron can lead to anemia and cause fatigue, cognitive impairment, and poor immune function. Animal-based foods provide heme iron, which is more bioavailable to humans, while plant-based foods typically contain less bioavailable non-heme iron. Edible insects vary in their iron content and may have heme or non-heme forms, depending on their diet. Edible insects have been proposed as a protein source that could address issues of food insecurity and malnutrition in low resource contexts; therefore, it is important to understand the bioavailability of iron from insect-based foods. In this study, we used Inductively Coupled Plasma and Mass Spectrometry (IPC-MS) and Caco-2 cell culture models to compare the soluble and bioavailable iron among five different lab-produced tempeh formulations featuring Tenebrio molitor (mealworm) with their non-fermented raw ingredient combinations. Finally, we compared the iron bioavailability of a mealworm tempeh with two sources of conventional beef (ground beef and sirloin steaks) and two commercially available plant-based meat alternatives. The results show that while plant-based meat alternatives had higher amounts of soluble iron, particularly in the Beyond Burger samples, the fermented mealworm-based tempeh had greater amounts of bioavailable iron than the other samples within the set. While all the samples presented varying degrees of iron bioavailability, all products within the sample set would be considered good sources of dietary iron. [ABSTRACT FROM AUTHOR]

Waste from non-degradable packaging materials poses a serious environmental risk and has led to interest in developing sustainable bio-based packaging materials. Sustainable packaging materials have been made from diverse naturally derived materials such as bamboo, sugarcane, and corn starch. In this study, we made a sustainable packaging film using chitosan extracted from the biomass of yellow mealworm (Tenebrio molitor) shell waste. The extracted chitosan was used to create films, cross-linked with citric acid (CA) and with the addition of glycerol to impart flexibility, using the solvent casting method. The successful cross-linking was evaluated using Fourier-Transform Infrared (FTIR) analysis. The CA cross-linked mealworm chitosan (CAMC) films exhibited improved water resistance with moisture content reduced from 19.9 to 14.5%. Improved barrier properties were also noted, with a 28.7% and 10.2% decrease in vapor permeability and vapor transmission rate, respectively. Bananas were selected for food preservation, and significant changes were observed over a duration of 10 days. Compared to the control sample, bananas packaged in CAMC pouches exhibited a lesser loss in weight because of excellent barrier properties against water vapor. Moreover, the quality and texture of bananas packaged in CAMC pouch remained intact over the duration of the experiment. This indicates that adding citric acid and glycerol to the chitosan structure holds promise for effective food wrapping and contributes to the enhancement of banana shelf life. Through this study, we concluded that chitosan film derived from mealworm biomass has potential as a valuable resource for sustainable packaging solutions, promoting the adoption of environmentally friendly practices in the food industry. [ABSTRACT FROM AUTHOR]

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

Over the past 2 decades, the potential of insects as food and feed has been recognized globally. Insects as feed ingredients can improve sustainability because of their lower greenhouse gas emissions and their potential to transform organic wastes into high-quality feed rich in nutrients. However, currently, the practical use of insects as food or feed is limited by the high costs of insect production. A great deal of effort is required to improve the rearing technology necessary to establish the principles of insect farming. Several insect species have become industrialized using existing methods of production. The most common industrialized insect species intended as feed and food include the yellow mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae); the house cricket, Acheta domesticus L. (Orthoptera: Gryllidae); the black soldier fly, Hermetia illucens L. (Diptera: Stratiomyidae); and the house fly, Musca domestica L. (Diptera: Muscidae). This review focuses on describing the existing rearing methods for these 4 insect species, which may provide a basis for future research to enhance insect production capabilities. [ABSTRACT FROM AUTHOR]

An in vivo trial was conducted to determine the apparent digestibility coefficients (ADCs) of insect meals for rainbow trout, Oncorhynchus mykiss. Rainbow trout (approximately 370 g ± 23 g, mean ± SD initial weight) were stocked 25 per tank into 400-liter tanks. Fish were fed a reference diet, or 1 of 5 test diets created by blending the reference diet in a 70:30 ratio (dry-weight basis) with menhaden fish meal (MFM), 2 house cricket (Acheta domesticus) meals (cricket A and cricket B), Galleria mellonella meal, and yellow mealworm (Tenebrio molitor) meal. Diets were assigned to 3 replicate tanks of fish and fed twice daily for 14 days prior to fecal collection. Ingredients, diets, and fecal matter were analyzed in duplicate for proximate, mineral, and amino acid composition. House cricket meals were 67.3% and 69.0% protein (CP) and 16.6% and 17.1% lipid (CL), for house cricket A and B, respectively. Yellow mealworm meal contained 56.5% CP and 27.7% CL, and G. mellonella larvae meal contained 32.5% CP and 54.2% CL. Protein ADCs were 78.9 for G. mellonella larvae meal, 78.0 for yellow mealworm meal, and 76.5 for house cricket A and not different from the MFM protein ADC of 76.6, while house cricket B protein ADC was 65.8 and was significantly lower than the MFM protein ADC (F  = 7.39; df = 4,14; P  = 0.0049). Together, these nutritional values suggest house crickets, and yellow mealworms show promise as alternative protein sources in salmonid feeds, with the potential of G. mellonella as an alternative lipid source. [ABSTRACT FROM AUTHOR]

The utilization of yellow mealworm, Tenebrio molitor (Linnaeus, Coleoptera: Tenebrionidae), for food and feed is gaining interest globally. However, its production is hindered by expensive commercial diets. This study assessed mealworm growth performance, survival, bioconversion, and nutritional composition when fed on wheat bran (WB) with different inclusion levels (25%, 50%, 75%, and 100%) of Irish potato waste (PW). Results indicated that mealworms fed on diets with 25%–75% PW had increased body length and 1–2 times higher weight gain compared to sole WB and PW diets. The survival rate was 93%–94% across all diets. Mealworms fed on WB had a feed conversion ratio of 3.26, while the efficiency of diet conversion increased with PW inclusion levels. Mealworms fed on diets with 75% PW inclusion had the highest crude fat (48%) and energy levels (598 kcal/100 g), while sole WB produced mealworms with the highest crude protein (55%). The acid detergent fiber achieved using 100% WB was 2- to 3-fold higher, but the crude fiber and neutral detergent fiber did not vary significantly. Considerable amounts of lysine (1.6–2 mg/100 g), methionine (0.5–0.7 mg/100 g), leucine (1.4–2 mg/100 g), and threonine (0.8–1 mg/100 g) were achieved in the mealworm larvae. Our findings revealed that cheap agricultural by-products could be successfully used for the mass production of mealworms, substantially contributing to reduced production costs. Further exploration of the nutrient-dense mealworm larvae for the development of novel food and feed products is crucial. [ABSTRACT FROM AUTHOR]

Insect manure or "frass" has emerged as an alternative nutrient source for alleviating the dependence on fossil fuel-based fertilizers, reducing food waste, and promoting food security. Yet, research on insect frass chemical composition is in its infancy. Here, we assessed the chemical properties of yellow mealworm (Tenebrio molitor L.) frass compared with poultry litter (PL). Insect frass was obtained from the National Biological Control Laboratory (NBCL; IF-L) and an insect-rearing company (IF-C). PL was collected from facilities in Arkansas (PL-AR) and North Carolina (PL-NC). Samples were analyzed for pH, electrical conductivity, macro- and micronutrients, heavy metals, pathogens, and indicator microorganisms. On average, insect frass had 43% and 47% higher C and N than PL, respectively (P  < 0.05). Considering a 5 mg/ha application rate, IF-C can supply 159 kg N/ha, twice the N supply of PL-AR (78 kg/ha). IF-L had a 53% higher P supply than PL-NC. Mean K, Ca, S, and micronutrient contents were higher in PL than in frass (P  < 0.05), whereas As, Cd, Cr, and Pb were nearly absent in frass. Chemical composition and pathogens in fertilizer sources were largely affected by insect-rearing substrate and supplements used in poultry and insect production. Insect frass utilized in this study had optimum C and N rates relative to PL, suggesting a promising soil amendment for improving soil health and C sequestration, thus contributing to sustainable agricultural intensification and reuse of food waste in circular economies. [ABSTRACT FROM AUTHOR]

The study aimed to be able to incorporate mealworm powder into yoghurts to increase protein content. Tenebrio molitor is gaining popularity as an alternative protein source. In the face of a growing human population and the associated challenges of sustainable nutrition, mealworms offer a promising solution. The mealworm is distinguished by its high protein content and for being rich in essential amino acids, vitamins, and minerals, making it a valuable component of diet. Among others, chemical composition, colour, texture, degree of syneresis, sensory analysis, and microbiological analysis were analysed. As expected, the addition of insect powder increased the protein content from 4.91% (0 TM) to 7.41% (5 TM). It also increased the fat content from 1.67% (0 TM) to 3.88% (5 TM). Furthermore, the addition of mealworm powder resulted in a significant change in the colour of the yoghurt to a darker colour, due to the dark brown colour of the powder. Food with added edible insects is increasingly available but is not always popular due to cultural and psychological barriers. Sensory analysis of yoghurts has shown that the more powder that is added, the less appealing the product becomes and the more bitter the taste. [ABSTRACT FROM AUTHOR]

As the most numerous group of animals on Earth, insects are found in almost every ecosystem. Their useful role in the environment is priceless; however, for humans, their presence may be considered negative or even harmful. For years, people have been trying to control the number of pests by using synthetic insecticides, which eventually causes an increased level of resistance to applied compounds. The effects of synthetic insecticides have encouraged researchers to search for alternatives and thus develop safe compounds with high specificity. Using knowledge about the physiology of insects and the functionality of compounds of insect origin, a new class of bioinsecticides called peptidomimetics, which are appropriately modified insect analogues, was created. One promising compound that might be successfully modified is the thirteen amino acid peptide alloferon (HGVSGHGQHGVHG), which is obtained from the hemolymph of the blue blowfly Calliphora vicinia. Our research aimed to understand the physiological properties of alloferon and the activity of its peptidomimetics, which will provide the possibility of using alloferon or its analogues in the pharmaceutical industry, as a drug or adjuvant, or in agriculture as a bioinsecticide. We used alloferon and its three peptidomimetics, which are conjugates of the native peptide with three unsaturated fatty acids with various chain lengths: caprylic, myristic, and palmitic. We tested their effects on the morphology and activity of the reproductive system and the embryogenesis of the Tenebrio molitor beetle. We found that the tested compounds influenced the growth and maturation of ovaries and the expression level of the vitellogenin gene. The tested compounds also influenced the process of egg laying, embryogenesis, and offspring hatching, showing that alloferon might be a good peptide for the synthesis of effective bioinsecticides or biopharmaceuticals. [ABSTRACT FROM AUTHOR]

Solanine (SOL) and chaconine (CHA) are glycoalkaloids (GAs) produced mainly by Solanum plants. These plant secondary metabolites affect insect metabolism; thus, they have the potential to be applied as natural plant protection products. However, it is not known which GA concentration induces physiological changes in animals. Therefore, the aim of this study was to perform a quantitative analysis of SOL and CHA in the larvae of Tenebrio molitor using LC‒MS to assess how quickly they are eliminated or metabolised. In this experiment, the beetles were injected with 2 μL of 10−5 M SOL or CHA solution, which corresponds to a dosage range of 0.12–0.14 ng/mg body mass. Then, 0.5, 1.5, 8, and 24 h after GA application, the haemolymph (H), gut (G), and the remainder of the larval body (FB) were isolated. GAs were detected in all samples tested for 24 h, with the highest percentage of the amount applied in the FB, while the highest concentration was measured in the H sample. The SOL and CHA concentrations decreased in the haemolymph over time, while they did not change in other tissues. CHA had the highest elimination rate immediately after injection, while SOL slightly later. None of the GA hydrolysis products were detected in the tested samples. One possible mechanism of the detoxification of GAs may be oxidation and/or sequestration. They may be excreted by Malpighian tubules, with faeces or with cuticles during moulting. The results presented are significant because they facilitate the interpretation of studies related to the effects of toxic substances on insect metabolism. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the impact of substituting a portion of feed with Tenebrio molitor (TM) and Elodea nuttallii (EN) on crayfish culture. A total of 270 crayfish (5.1 ± 0.4 g) were fed three different diet combinations (A: 100% feed; B: 80% feed + 10% TM + 10% EN; C: 75% feed + 15% TM + 10% EN) for 12 weeks. The findings demonstrated that group C had an important beneficial impact on the growth performance of crayfish. This was evidenced by a rise in digestive enzyme activity (trypsin, lipase, and cellulase) in the intestinal and hepatopancreas, as well as an upregulation in the expression of growth-related genes (ghsr, igfbp7, mhc, mlc1, mef2, and pax7) in the muscle. Furthermore, the assessment of the flesh quality of crayfish muscle in group C was conducted. The findings indicated a significant increase (p < 0.05) in the energy value (moisture, crude protein, and crude lipid) within the muscle. The levels of delicious amino acids (Glu, Ala, Ser, Gly, and Tyr) and polyunsaturated fatty acids (ARA, DHA) were enhanced, resulting in an improved nutritional profile and flavor of the muscle while maintaining the Σn-3/Σn-6 ratio. The remodeling of the intestinal microbiota (abundance of Proteobacteria and ratio of Firmicutes/Bacteroidota bacteria) also revealed improved growth performance. Additional research is necessary to ascertain whether excessive use of TM or EN feed substitution can have negative effects on crayfish culture. [ABSTRACT FROM AUTHOR]

This study aimed to assess the effect of the addition (2%) of soybean protein (SP) and insect flours derived from house crickets (Acheta domesticus, HCF) and yellow mealworm (Tenebrio molitor, YMF) in cooked sausages. The technological characteristics of the batter, the chemical composition of the sausages, their technological traits and lipid stability during refrigerated storage, as well as their sensory properties, were investigated. The SP, HCF and YMF batters displayed higher pH (p = 0.0025) and stability (p < 0.0001) but a darker colour (p < 0.0001) than the control samples. The addition of SP increased the plasticity of the batter (p = 0.0017), while YMF decreased its structural strength (p = 0.0274). Higher pH and darker colour were detected in SP-, HCF- and YMF-containing sausages; however, the effect of the alternative proteins depended on the duration of storage. The plasticity decreased in the insect-containing sausages (p = 0.0010) and increased over time (p = 0.0136), whereas the elasticity was lower in the YMF group (p < 0.0001). The protein and fat contents were higher (p < 0.0001) in the sausages containing alternative protein. TBARS content decreased over time in these groups. The HCF and YMF sausages received lower scores for their appearance, colour, texture, flavour and taste, suggesting the need for further technological interventions to make such products more attractive to consumers. [ABSTRACT FROM AUTHOR]

Historically, males have frequently been portrayed as the manipulative and deceptive gender, while females are often seen as adopting a coy and passive role. In this context, it is proposed that males use a terminal investment strategy, misleading females about their true poor condition, while females passively opt to mate with these deceptive males. However, we hypothesize that females in suboptimal condition may also engage in a terminal investment strategy by mimicking or enhancing their attractiveness to match that of females in better conditions. We studied this hypothesis in Tenebrio molitor, by subjecting females to three varying doses of lipopolysaccharides of Escherichia coli (LPS; 0.25, 0.5, or 1 mg ml-1), or three doses of the pro-oxidant Paraquat (PQ; 20, 40 or 80 mM), and subsequently assessing their survival and attractiveness to males. The LPS treatments and 20 mM of PQ had no significant effect on the survival or attractiveness of the females. However, females treated with 40 or 80 mM PQ survived fewer days compared to the control group. Those injected with 40 mM were more attractive than their control counterparts, while those treated with 80 mM were less attractive. Since the identical doses of LPS, which induce terminal investment in males, had no effect on females, we suggest sexual dimorphism in terminal investment. Furthermore, similar to males, if the stressor reaches a sufficiently high level, the signal becomes honest. These findings highlight how the quantity of stressors influences support for the terminal investment strategy in both males and females. Notably, this study challenges prevailing notions regarding gender roles in sexual selection, indicating that females, not just males, conceal their poor condition to attract mating partners. [ABSTRACT FROM AUTHOR]

Simple Summary: Due to the growing world population, the sustainability of food and feed sources with high nutritional value has become a crucial issue. In this scenario, insects can constitute a low-impact protein source with high nutritional value. The growth and nutrient composition of insects are potentially influenced by rearing conditions, particularly by the selected growth substrate. In this study, we evaluated the chemical and functional characteristics of Tenebrio molitor larvae reared on different growth substrates: a traditional wheat bran substrate and an innovative substrate consisting of wheat bran supplemented with chestnut shell, a by-product of the chestnut agro-industrial chain. The results showed that the innovative growth substrates positively influenced the insects' survival suggesting a beneficial effect on larval health. The enrichment of the growth substrate with chestnut shell modified the protein and amino acid profile of insect meals, possibly indicating a shift in their metabolism. In addition, insect meals obtained from larvae reared on chestnut-shell-enriched substrate exhibited higher antibacterial and antioxidant activity, suggesting a potential beneficial effect when included in animal feed. Our results showed positive outcomes related to the design of innovative strategies for insect rearing, enriching larvae meal with beneficial health properties in line with sustainability and One Health principles. Tenebrio molitor larvae represent a sustainable protein source for food and feed. The aim of this study was to evaluate the supplementation of chestnut shell, a by-product of the agro-industrial chain, in growth substrates for T. molitor larvae rearing. Seven-week-old larvae were reared on three different growth substrates: the control group (CTRL) was fed wheat bran, treatment group one was fed wheat bran supplemented with 12.5% w/w chestnut shell (TRT1), and treatment group two was fed wheat bran supplemented with 25% w/w chestnut shell (TRT2). Larval weight, substrate consumption, and mortality were recorded weekly. After 14 days, insect meals were produced for bromatological and colorimetric analysis, and bacterial inhibition activity assay using a microdilution method. The amino acid profile of insects was determined using quantitative nuclear magnetic resonance spectroscopy. Our results showed a lower feed conversion ratio and higher larval survival rate % in TRT2 compared to CTRL (p < 0.05). Proteins and lipids of TRT2 were higher than other groups (p < 0.05). Important differences were observed in the amino acid profile of TRT1 and TRT2 compared to CTRL (p < 0.05). TRT1 and TRT2 showed higher E. coli inhibitory activity than CTRL (p < 0.05). In conclusion, chestnut shell supplementation improved the survival and functional characteristics of larvae and likely impacted the insects' metabolism. [ABSTRACT FROM AUTHOR]

Simple Summary: Substances that are absent in the biosphere, but have been synthesized by humans in recent decades, are called xenobiotics. They are released into the environment in large volumes and can actively affect invertebrates and their intestinal parasites or commensals. Such an influence of xenobiotics on parasite–host relationships remained poorly studied until recently. In this article, the host–parasite system "larva of Tenebrio molitor—Gregarina polymorpha" under the influence of 24 organic compounds is investigated. We found that several substances increase the mortality of T. molitor larvae, and substances such as diphenyl ether, benzyl alcohol, catechol, and 3-aminobenzoic acid, on the contrary, reduce the number of parasites (G. polymorpha) without significantly affecting host insects (larva of T. molitor). Thus, various xenobiotics can destroy the parasite–host relationship, having a greater effect on the host in some cases and on the parasite in other cases. Environmental contamination with xenobiotics affects organisms and the symbiotic relations between them. A convenient object to study relationships between parasites and their hosts is the host–parasite system "Tenebrio molitor Linnaeus, 1758 (Coleoptera, Tenebrionidae)—Gregarina polymorpha (Hammerschmidt, 1838) Stein, 1848 (Eugregarinorida, Gregarinidae)". For this experiment, we took 390 T. molitor larvae and 24 organic compounds. Groups of mealworms, 15 in each, were subjected to those compounds for 10 days. Then, we recorded the vitality of both the larvae of T. molitor and G. polymorpha. To assess how G. polymorpha had affected the hosts' wellbeing, we looked for changes in the larvae's body mass and compared them to the number of gregarines in their intestines. The vitality of the larvae was inhibited by cyclopentanol and 2-naphthol. The intensity of gregarine invasion was reduced by diphenyl ether, benzyl alcohol, catechol, and 3-aminobenzoic acid. No effect on the number of gregarines was produced by 3,4,5-trihydroxybenzoic acid, cyclohexanemethanol, phenol, benzalkonium chloride, maleic anhydride, cyclohexanol, resorcin, benzoic acid, 2-methylfuran, terpinen-4-ol, 1-phenylethylamine, dibutyl phthalate, 3-furancarboxylic acid, 5-methyl furfural, 6-aminohexanoic acid, succinic anhydride, o-xylene, and benzaldehyde. In the infected T. molitor individuals, the mean number of G. polymorpha equaled 45 specimens per host. The groups of smaller mealworms had fewer gregarines. Positive correlation was seen between growth rates of T. molitor larvae and the intensity of invasion by gregarines. [ABSTRACT FROM AUTHOR]

Insects are of great interest as novel sources of alternative proteins and biologically active compounds, primarily anticancer agents. Protein-rich insect larval hemolymph is a prospective candidate for pharmaceutical and food industry-related research. In this study, selected biochemical properties and cell toxicity of larval hemolymph from two mealworm species, Tenebrio molitor and Zophobas morio, were analyzed. Total proteins and carbohydrates, antioxidant capacity, and the level of lipid peroxidation were determined. Human cancer (U-87) and normometabolic (MRC-5) cells were treated with different concentrations of larval hemolymph proteins, and the effects on cell viability were assayed 24, 48, and 72 h after treatments. Z. morio hemolymph was shown to be richer in total proteins, showing a higher antioxidant capacity and lipid peroxidation level than T. molitor hemolymph, which was richer in total carbohydrates. Cytotoxicity assays showed that T. molitor and Z. morio hemolymphs differently affect the viability of U-87 and MRC-5 cells in cell type-, dose-, and time-dependent manners. Hemolymph from both species was more cytotoxic to U-87 cells than to MRC-5 cells, which was particularly prominent after 48 h. Additionally, a more potent cytotoxic effect of Z. morio hemolymph was observed on both cell lines, likely due to its higher antioxidant capacity, compared to T. molitor hemolymph. [ABSTRACT FROM AUTHOR]

Foods enriched with insects can potentially prevent several health disorders, including cardiovascular diseases, by reducing inflammation and improving antioxidant status. In this study, Tenebrio molitor and Gryllus assimilis were selected to determine the effect on the development of atherosclerosis in ApoE/LDLR−/− mice. Animals were fed AIN-93G-based diets (control) with 10% Tenebrio molitor (TM) and 10% Gryllus assimilis (GA) for 8 weeks. The nutritional value as well as antioxidant activity of selected insects were determined. The lipid profile, liver enzyme activity, and the fatty acid composition of liver and adipose tissue of model mice were evaluated. Quantitative analysis of atherosclerotic lesions in the entire aorta was performed using the en face method, and for aortic roots, the cross-section method was used. The antioxidant status of the GA cricket was significantly higher compared to the TM larvae. The results showed that the area of atherosclerosis (en face method) was not significantly different between groups. Dietary GA reduced plaque formation in the aortic root; additionally, significant differences were observed in sections at 200 and 300 µm compared to other groups. Furthermore, liver enzyme ALT activity was lower in insect-fed groups compared to the control group. The finding suggests that a diet containing edible insect GA potentially prevents atherosclerotic plaque development in the aortic root, due to its high antioxidant activity. [ABSTRACT FROM AUTHOR]

Tenebrio molitor gained recognition for its nutritional value for human and/or animal consumption, its utilization as a model species in research studies, and its ability to degrade many types of plastics. However, it is a stored-product pest infesting a wide range of commodities in storages. The impact of temperature on the development of T. molitor was evaluated, taking into account confounding effects of other covariates such as developmental stage and larval instar. The time for larval development was longer at 22.5 °C than at 25 °C, 27.5 °C, and 30 °C. Tenebrio molitor spent most of its lifetime as larva > egg, pupa, since the outcome of our model inference resulted that the estimated probability of longer duration in larvae is approximately 25.5 times higher compared to eggs and pupae, and passed through 26 instars before it became pupa. Duration of larval instars increased significantly up to L22 and then decreased until L26. Among larval instars, L22 exhibited the highest developmental duration while the lowest was recorded for L1. These findings enhance the knowledge about the biology of T. molitor, leading to precise decisions for its successful culture. [ABSTRACT FROM AUTHOR]

Fresh insects in the form of paste can be conveniently utilised for further food preparations and processing at both household and industrial levels. However, due to the high moisture content and particular physiochemical properties, insect paste is highly unstable and susceptible to significant quality changes during storage. The aim of this study was to evaluate the microbiological and physicochemical stability of high hydrostatic pressure (HHP) treated Tenebrio molitor paste stored at different modified atmosphere packaging (MAP) conditions. Three different MAP compositions and normal air were applied. pH, colour, lipid oxidation, fluorescence spectra and microbiological parameters were evaluated along 28 days of refrigerated storage (4 °C). HHP treatment (600 MPa, 5 min) resulted in a significant reduction in mesophilic aerobic bacteria, anaerobic bacteria and yeast and mould counts. Growth of mesophilic aerobic and anaerobic bacteria remained low for 7 days at 4 °C, regardless of the applied atmospheric conditions, while high microbial growth was observed after 14 days without any significant differences among the treatments. The number of mesophilic spores remained low throughout the entire storage period when MAP was applied, whereas high values were observed after 28 days in the samples stored in normal air. Listeria monocytogenes, Salmonella spp. and Escherichia coli were not detected in any of the samples, nor in the raw paste. Lipid oxidation was initiated by the HHP treatment and displayed an increasing trend during the storage period, although the oxidation levels were lower in the MAP without oxygen compared to the normal air packaging and in MAPs with high O2 or CO2 levels. Fluorescence spectroscopy showed high potential for rapid detection of chemical changes in insect products during processing and storage. Overall, this study showed that MAP can effectively be applied on HHP-treated insect paste to extend its storage period. Absence of oxygen has been identified as a crucial factor in achieving reduced microbial growth and ensuring high physiochemical quality of the final product. Therefore, the present research offers a valuable insight for creating new edible insect-based products within the food industry. [ABSTRACT FROM AUTHOR]

There is a need for alternatives or complements to synthetic fertilizers to enhance agricultural sustainability. Applying organic amendments can play a significant role in this. Insect droppings show high potential, though studies evaluating their agronomic value have only recently begun to emerge. This study compared black soldier fly (Hermetia illucens L.) and mealworm (Tenebrio molitor L.) larvae frass with another organic amendment (Nutrimais) derived from composting forestry, agro-industrial, and domestic waste. The experiment also included ammonium nitrate at two rates [the same as the organic amendments, 50 kg ha−1 nitrogen (N) (FullR), and half that rate (HalfR)] and an unfertilized control. The study spanned two growth cycles of lettuce (Lactuca sativa L.) grown in pots, followed by unfertilized oats (Avena sativa L.) to assess the residual effects of the fertilizing treatments. Mealworm larvae frass mineralized rapidly, with an apparent N recovery of 37.4% over the two lettuce growth cycles, indicating its high availability to soil heterotrophic microorganisms. The average dry matter yield (DMY) of lettuce was the highest among all treatments (12.8 and 9.8 g plant−1 in the first and second lettuce cycles), even compared to the FullR treatment (12.2 and 7.8 g plant−1), though without significant differences. Although mealworm larvae frass exhibited a high mineralization rate, the DMY cannot be attributed solely to N supply, as plants in the FullR treatment showed better N nutritional status. Mealworm larvae frass provided strong evidence of a plant biostimulant effect, not explained by the variables measured in this study. Black soldier fly larvae frass exhibited typical behavior of a moderately reactive organic amendment, while Nutrimais showed low reactivity, with a near-neutral mineralization/immobilization balance. The results suggest mealworm larvae frass is recommended for early maturing vegetable crops, whereas Nutrimais appears more suitable for perennial crops with low short-term nutrient requirements. [ABSTRACT FROM AUTHOR]

Background: Insects encounter various environmental stresses, in response to which they generate reactive oxygen species (ROS). Superoxide dismutase (SOD) is an antioxidant metalloenzyme that scavenges superoxide radicals to prevent oxidative damage. Objective: To investigate expressions of SODs under oxidative stress in Tenebrio molitor. Methods: Here, we investigated the transcriptional expression of SODs by pesticide and heavy metals in Tenebrio moltior. First, we searched an RNA-Seq database for T. molitor SOD (TmSOD) genes and identified two SOD isoforms (TmSOD1-iso1 and iso2). We examined their activities under developmental stage, tissue-specific, and various types (pesticide and heavy metal) of oxidative stress by using qPCR. Results: Our results revealed two novel forms of TmSODs. These TmSODs had a copper/zinc superoxide dismutase domain, active site, Cu2+ binding site, Zn2+ binding site, E-class dimer interface, and P-class dimer interface. TmSODs (TmSOD1-iso1 and iso2) were expressed in diverse developmental phases and tissues. Pesticides and heavy metals caused an upregulation of these TmSODs. Conclusion: Our findings suggest that the two TmSODs have different functions in T. molitor, providing insights into the detoxification ability of T. molitor. [ABSTRACT FROM AUTHOR]

Mealworms are one of the most economically important insects in large-scale production for human and animal nutrition. Densoviruses are highly pathogenic for invertebrates and exhibit an extraordinary level of diversity which rivals that of their hosts. Molecular, clinical, histological, and electron microscopic characterization of novel densovirus infections is of utmost economic and ecological importance. Here, we describe an outbreak of densovirus with high mortality in a commercial mealworm (Tenebrio molitor) farm. Clinical signs included inability to prehend food, asymmetric locomotion evolving to nonambulation, dehydration, dark discoloration, and death. Upon gross examination, infected mealworms displayed underdevelopment, dark discoloration, larvae body curvature, and organ/tissue softness. Histologically, there was massive epithelial cell death, and cytomegaly and karyomegaly with intranuclear inclusion (InI) bodies in the epidermis, pharynx, esophagus, rectum, tracheae, and tracheoles. Ultrastructurally, these InIs represented a densovirus replication and assembly complex composed of virus particles ranging from 23.79 to 26.99 nm in diameter, as detected on transmission electron microscopy. Whole-genome sequencing identified a 5579-nucleotide-long densovirus containing 5 open reading frames. A phylogenetic analysis of the mealworm densovirus showed it to be closely related to several bird- and bat-associated densoviruses, sharing 97% to 98% identity. Meanwhile, the nucleotide similarity to a mosquito, cockroach, and cricket densovirus was 55%, 52%, and 41%, respectively. As this is the first described whole-genome characterization of a mealworm densovirus, we propose the name Tenebrio molitor densovirus (TmDNV). In contrast to polytropic densoviruses, this TmDNV is epitheliotropic, primarily affecting cuticle-producing cells.

Preventing disease and maintaining the health of the elderly are crucial goals for an aging population, with obesity and immune function restoration being of paramount importance. Obesity, particularly visceral obesity characterized by excessive fat accumulation around the abdominal organs, is linked to chronic conditions such as diabetes, hypertension, cardiovascular diseases, and immune dysfunction. Globally, obesity is considered a disease, prompting significant research interest in its treatment. Therefore, it is essential to explore potential therapeutic and preventive strategies to address obesity and the decline in immune function brought about by aging. Tenebrio molitor larvae (TML), commonly known as ‘mealworms,’ are rich in unsaturated fatty acids, including oleic and linoleic acids, and essential amino acids, such as isoleucine and tyrosine. In this study, we aimed to investigate the effects of the consumption of TML oil and mealworm fermented extract (MWF-1) on obesity and immunological changes in aged obese mice. Our data showed reduced body fat in 23-week-old C57BL/6 mice fed processed TML products for 6 weeks. Additionally, the characteristically high levels of serum triglycerides decreased by treating with TML oil. The immune responsiveness results confirmed an increase in B cells by treating with MWF-1, while cytokine levels (interferon-gamma, tumor necrosis factor-alpha, interleukin-2, and −6) were restored to levels similar to young mice. These results suggest that TML oil and MWF-1 are promising dietary supplements for addressing obesity and restoring immune function.

Production of insects as animal protein source has increased recently but the information on nutrient requirements for optimal growth is still scarce. The effects of carbohydrate composition of the substrate on growth performance and chemical composition of Tenebrio molitor larvae were evaluated. Three substrates based on different proportions of wheat grain, wheat bran and soybean meal were formulated to contain three starch to neutral detergent fibre (NDF) ratios: low (1.0; LSF), medium (1.2; MSF) and high (1.4; HSF), at a common N level. Each treatment was replicated five times in trays containing 23 g of substrate and 40 larvae (2.95 mg average weight) each. Weight of the larvae and the substrate remaining per tray were weekly monitored from 21 to 49 days of experiment and larvae within the same experimental treatment were pooled at the end of the trial for chemical analysis. No treatment differences were observed for larval growth or mortality (p > 0.05). The starch to NDF ratio in the substrate did not affect larval composition in terms of dry matter, nitrogen or ether extact contents or the amino acid composition of the protein fraction. Within the range of ingredient composition considered, the starch to NDF ratio in the substrate has a minor influence on growth performance and chemical composition of T. molitor larvae.

In the last decade, great attention was placed on insects to increase feed-food production without negatively affecting the environment. In this research study we compare three different insect species (Alphitobius diaperinus, Tenebrio molitor, and Zophobas morio - lesser mealworm, mealworm, and superworm), fed the same substrate, on the chemical composition, fatty acid (FA) profile, antioxidant content, and microbiological loads. Superworm larvae show higher dry matter (38.06%) and ether extract (17.65%) contents followed by mealworm and lesser mealworm larvae (32.77%, 27.11% and 13.86%, 10.78%, respectively). No differences were detected in the crude protein and ash content. Superworm larvae showed the highest content of SFA (41.43% vs 39.16% and 28.52% of total FA, in lesser mealworm and mealworm, respectively), while mealworm larvae were the richest in MUFA (40.61% vs 34.38% and 32.98% of total FA, in super and lesser mealworm, respectively) and PUFA (29.05% vs 22.29% and 21.21% of total FA in super and lesser mealworm, respectively). Anyhow, in all the larvae the three more representative fatty acids were oleic acid, linoleic acid, and palmitic acid. The least beneficial ratio of n-6/n-3 was detected in Alphitobius diaperinus larvae (53.84), while Zophobas morio larvae showed the healthier ratio (16.04). The antioxidant contents determination revealed a linkage to the fatty acids content. Low differences were determined in microbiological loads of the larvae. The characteristics of the insects revealed the great potential of these three species highlighting the capacities to respond to different requests derived from the food and feed sectors.Highlights Superworm larvae showed the highest content of SFA Mealworms larvae were the richest in PUFA The fat-soluble vitamin content followed the fatty acids content

Numerous insect species and their associated microbial pathogens are exposed to elevated CO2 concentrations in both artificial and natural environments. However, the impacts of elevated CO2 on the fitness of these pathogens and the susceptibility of insects to pathogen infections are not well understood. The yellow mealworm, Tenebrio molitor, is commonly produced for food and feed purposes in mass-rearing systems, which increases risk of pathogen infections. Additionally, entomopathogens are used to control T. molitor, which is also a pest of stored grains. It is therefore important to understand how elevated CO2 may affect both the pathogen directly and impact on host-pathogen interactions. We demonstrate that elevated CO2 concentrations reduced the viability and persistence of the spores of the bacterial pathogen Bacillus thuringiensis. In contrast, conidia of the fungal pathogen Metarhizium brunneum germinated faster under elevated CO2. Pre-exposure of the two pathogens to elevated CO2 prior to host infection did not affect the survival probability of T. molitor larvae. However, larvae reared at elevated CO2 concentrations were less susceptible to both pathogens compared to larvae reared at ambient CO2 concentrations. Our findings indicate that whilst elevated CO2 concentrations may be beneficial in reducing host susceptibility in mass-rearing systems, they may potentially reduce the efficacy of the tested entomopathogens when used as biological control agents of T. molitor larvae. We conclude that CO2 concentrations should be carefully selected and monitored as an additional environmental factor in laboratory experiments investigating insect-pathogen interactions. [ABSTRACT FROM AUTHOR]

Abstract Background This study aimed to evaluate the effects of Hermetia illucens (Black soldier fly-BSF) and Tenebrio molitor (Yellow mealworm-YMW) live larvae as a new nutritional model on duck’s gut health, considering gut histomorphometry, mucin composition, cytokines transcription levels, and microbiota. A total of 126, 3-days-old, females Muscovy ducks were randomly allotted to three dietary treatments (6 replicates/treatment, 7 birds/pen): (i) C: basal diet; (ii) BSF: C + BSF live larvae; (iii) YMW: C + YMW live larvae. BSF and YMW live larvae were administered on top of the basal diet, based on the 5% of the expected daily feed intake. The live weight, average daily gain, average daily feed intake and feed conversion ratio were evaluated for the whole experimental period. On day 52, 12 ducks/treatment (2 birds/replicate) were slaughtered and samples of duodenum, jejunum, ileum, spleen, liver, thymus and bursa of Fabricius were collected for histomorphometry. Mucin composition was evaluated in the small intestine through histochemical staining while jejunal MUC-2 and cytokines transcription levels were evaluated by rt-qPCR. Cecal microbiota was also analyzed by means of 16 S rRNA gene sequencing. Results Birds’ growth performance and histomorphometry were not influenced by diet, with a proximo-distal decreasing gradient from duodenum to ileum (p

Monopterus albus is one of China's renowned and superior aquaculture species, with its seedlings mainly sourced from wild capture. One of the bottlenecks in M. albus aquaculture is the high mortality rate and low feeding initiation rate from stocking wild fry to the initiation of feeding. In production, trash fish is commonly used to wean M. albus juveniles onto feeding. In this study, we introduced three other natural feeds, earthworms (EW), yellow mealworms (YMW), and fly maggots (FM), with frozen trash fish (TF) serving as the control group, to evaluate the effects of these four natural feeds on the survival rate, feeding initiation, antioxidant enzymes activity, and body composition of M. albus juveniles under recirculating water aquaculture conditions. The experiment comprised four treatments, each with three replicates. Each replicate consisted of stocking 150 M. albus juveniles weighing 10.02 ± 0.89 g in size, raised for 5 weeks. The survival rate of the YMW group was 73.33%-85.33%, which was significantly higher than that of the other three bait groups (p < 0.05). The four bait groups showed no significant differences in final body weight and specific growth rate (SGR) (p > 0.05). The EW group showed the highest final body weight, with an average SGR of 2.73, whereas the YMW group had an average SGR of 1.87. The average daily feeding amount was significantly higher in EW and YMW groups than in the other two groups (p < 0.05). The percentage of feeding amount to fish weight in the EW group reached 7.3% in the fifth week. After 5 weeks of cultivation, NO2--N content was significantly higher in the waters of the TF and EW groups than in the waters of the FM and YMW groups (p < 0.05), there was no significant difference in TAN content among the treatment groups (p > 0.05). Liver malondialdehyde content was significantly higher in the TF group than in the other bait groups (p < 0.05). GSH-Px activity was significantly higher in the EW group than in the FM group and YMW group. No significant differences in SOD and CAT activity and T-AOC were observed among the bait groups (p > 0.05). The increase in crude protein content was significantly higher in the TF group than in the FM group, but the increase in crude ash content was significantly lower in the TFgroup. In conclusion, Tenebrio molitor could potentially serve as one of the alternative feeds during the initial stages of M. albus juveniles stocking. [ABSTRACT FROM AUTHOR]

Here, a comprehensive cytogenetic analysis of three species of tenebrionid beetles: Alphitobius diaperinus, Tenebrio molitor and Zophobas morio, is presented. This paper also contains a review of the cytogenetic information for each species and adds new data. The male karyotype of both T. molitor and Z. morio is 2n = 18 + Xyp, with large heterochromatic blocks in the pericentromeric regions of all chromosomes. The male A. diaperinus karyotype is 2n = 18 + X0, also with heterochromatic pericentromeric regions in all chromosomes. The location of the nucleolar organizer regions (NORs) differs in each species: in A. diaperinus, it is located on the smallest pair of autosomes, while in T. molitor, it is on two pairs of autosomes and both sex chromosomes. In contrast, it is exclusively located on the X chromosome in Z. morio. Telomere analysis revealed that all species have TCAGG repeats in their telomeres, but lack the canonical TTAGG insect telomeric motif. In addition, a study of the distribution of satellite DNA and composition revealed that each species has a main satellite DNA family forming the pericentromeric heterochromatin. Fluorescence in situ hybridization of each of these satellites did not produce hybridization signal in the other two species, indicating a divergence in repetitive DNA composition among them. This study adds to the understanding of chromosomal organization, heterochromatin distribution and repetitive DNA dynamics in tenebrionid beetles and sheds light on their cytogenetic diversity and evolutionary significance. [ABSTRACT FROM AUTHOR]

A degradation mechanism of polystyrene (PS) in mealworms reared on expanded PS (EPS) was investigated by its decrease in molecular weight and change in chemical structure. A 33% decrease in molecular weight was observed for the digested PS in the frass after 1 week of feeding to mealworms. The FT-IR and py-GC/MS spectra of the digested PS showed radical oxidative reactions taking place in the mealworm body. The presence of hydroperoxide, alcohol and phenol groups was confirmed, and dimer fragments of styrene with quinone and phenol groups were obtained. The decrease in molecular weight and the alternation of benzene rings indicated that autoxidation and quinonization via phenolic intermediates occurred simultaneously in the mealworm body. The survival rate of mealworms reared on EPS was higher than that of starved worms, indicating that EPS was a nutrient source. However, no weight gain was observed in mealworms fed EPS alone. Comparison with the mixed diets with bran or urethane foams (PU) indicated that protein, phosphorus and magnesium components absent from EPS were required for mealworm growth. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the response of grass carp feeding, growth, and antioxidant capacity to dietary protein content when using yellow mealworms as the main protein source. With yellow mealworms as the main protein source, 6 experimental diets with different protein level (25.25%, 29.43%, 33.38%, 37.96%, 41.40% and 45.70%, named as TMF1, TMF2, TMF3, TMF4, TMF5, and TMF6 group, respectively) were designed. The grass carp juveniles with initial weight of (3.63± 0.01) g were fed with the experimental diets for 8 weeks. The results are as follows: ① the specific growth rate, weight gain rate, and protein efficiency ratio of grass carp decreased with the increase of yellow mealworm addition, all of which were significantly higher in TMF1 group than those in FMF4, TMF5 and TMF6 groups (P<0.05), the feeding rate showed a first increasing and then decreasing trend, while the survival rate and feed efficiency were not affected (P>0.05) by dietary mealworm addition. ② Different levels of dietary yellow mealworm inclusion had no significant effect on the condition factor and percentage of eviscerated weight of grass carp (P>0.05), but the hepatopancreas somatic index and mesenteric fat index of grass carp increased first and then decreased with the increase of yellow mealworm content. The mesenteric fat index of TMF2 group was significantly higher than that of other groups (P<0.05). ③ The total protein, content in the serum of grass carp first increased and then decreased with the increase of yellow mealworms content. And the total protein content in the TMF4 group was significantly higher than those in other groups (P<0.05), but was not different from that in TMF3 group. The serum cholesterol content in TMF1 group was significantly lower than that in other groups, and the activity of aspartate aminotransferase in TMF4 and TMF6 group was significantly higher than that in TMF1, TMF2 and TMF3 group (P<0.05), while the content of triglyceride decreased significantly with the increase of the amount of yellow mealworms, which was significantly higher in the TMF1 group than those in TMF5 and TMF6 groups (P<0.05). The serum glucose in TMF1 group was significantly higher than that in TMF2 group (P<0.05), no other difference in serum glucose was observed. ④ The activities of SOD, T-AOC, MDA and GSH-Px in grass carp hepatopancreas decreased with the increase of yellow mealworms, and the GSH-Px activity as well the MDA content were significantly higher in TMF1 group than in other groups (P<0.05). However, the CAT activity did not vary significantly among groups (P>0.05). This study shows that the protein level of grass carp diet should not be more than 25.25% when yellow mealworms was used as the main protein source, and the increase of dietary yellow mealworms inclusion would depress the feeding, growth, and antioxidant performance of grass carp. [ABSTRACT FROM AUTHOR]

We characterized an alkaline chymotrypsin-like serine peptidase from the yellow mealworm Te-nebrio molitor with a non-canonical substrate-binding subsite for its possible application as a component (an additive) in various biological products. The enzyme was obtained as a recombinant preparation. Purification was carried out using affinity chromatography on Ni2+-NTA agarose. The specificity constants (kcat/KM) for the chymotrypsin substrates, Glp-AAF-pNA, Suc-AAPF-pNA, and Ac-Y-pNA, were 7, 4.2, and 0.9 (µM min)–1, respectively. The optimum of the proteolytic activity was observed at pH 9.0. The enzyme was stable in the alkaline pH range, and in the presence of BSA, also in the acidic region. Peptidase was inhi-bited by synthetic inhibitors such as PMSF, TPCK, and chymostatin, while EDTA, E-64, and pepstatin had no effect on the enzyme activity. The purified enzyme showed high stability over time in the presence of BSA. The short life cycle of the insect and the production of a large number of peptidases in the midgut with high catalytic activity and stability can make T. molitor an excellent alternative source of industrially important enzymes for application as components (additives) in various biological products (e.g., stain removers, detergents, etc.). [ABSTRACT FROM AUTHOR]

Among prey functional traits, mobility plays a major role, affecting its actual availability to predators: a moving animal is usually more susceptible of being detected than immobile or inanimate prey. With the aim of investigating the role played by visual cues and preference for moving vs. immobile prey in a widespread, generalist species, common wall lizard Podarcis muralis , we used two different experimental approaches. First, we assessed the preference of tested individuals for either one active or one still mealworm (Tenebrio molitor), symmetrically hung in a T-shaped arena (single prey preference experiment); secondly, each individual was left free to interact for 20 min with two closed boxes containing 10 moving or still worms, kept at opposite sides of a rectangular arena (multiple prey experiment). Our results showed that lizards consistently selected the moving stimuli when no chemical prey-borne cues were available. When choosing between single prey, we found inter-individual variation in both lizard preference for moving or still prey and latency to attack, which may depend on either microhabitat-related difference in foraging tactics or personality traits. [ABSTRACT FROM AUTHOR]

A class of proteins known as antifreeze proteins (AFPs) appear in some organisms, allowing them to survive in low-temperature environments. These AFPs irreversibly adsorb to the surfaces of ice crystals and reduce the freezing temperature without significantly affecting the equilibrium melting point. Ice crystal growth is inhibited in the temperature gap between the melting point and the non-equilibrium freezing point, referred to as thermal hysteresis (TH). The irreversible adsorption of AFPs on the surfaces of ice crystals has been questioned because it is not consistent with the concentration dependence of the TH activity obtained from experimental studies. In this study, based on adsorption–inhibition theory, a random sequential adsorption model of AFPs was used to investigate the irreversible adsorption of hyperactive AFPs that have been found in the yellow mealworm beetle Tenebrio molitor (TmAFPs). The occupied fractions covered by TmAFP on ice crystal surfaces were obtained. The time and concentration dependence of the TH activity of TmAFP was analyzed. The theoretical results obtained from this model were consistent with reported experimental data. This work provides ideas and a theoretical basis for understanding the TH activity during the irreversible adsorption of AFPs. [ABSTRACT FROM AUTHOR]

Serine peptidases (SPs) of the chymotrypsin S1A subfamily are an extensive group of enzymes found in all animal organisms, including insects. Here, we provide analysis of SPs in the yellow mealworm Tenebrio molitor transcriptomes and genomes datasets and profile their expression patterns at various stages of ontogeny. A total of 269 SPs were identified, including 137 with conserved catalytic triad residues, while 125 others lacking conservation were proposed as non-active serine peptidase homologs (SPHs). Seven deduced sequences exhibit a complex domain organization with two or three peptidase units (domains), predicted both as active or non-active. The largest group of 84 SPs and 102 SPHs had no regulatory domains in the propeptide, and the majority of them were expressed only in the feeding life stages, larvae and adults, presumably playing an important role in digestion. The remaining 53 SPs and 23 SPHs had different regulatory domains, showed constitutive or upregulated expression at eggs or/and pupae stages, participating in regulation of various physiological processes. The majority of polypeptidases were mainly expressed at the pupal and adult stages. The data obtained expand our knowledge on SPs/SPHs and provide the basis for further studies of the functions of proteins from the S1A subfamily in T. molitor. [ABSTRACT FROM AUTHOR]

Edible insects such as the mealworm (Tenebrio molitor) are emerging as an alternative animal protein resource to meet the needs of a growing global population. However, despite the successful mass production of edible insects, research into practical compound feed for edible insect is still insufficient. In this study, mealworms were fed wheat bran, layer chicken feed mash, chicken pellets and cattle pellets for 1 to 36 days, after which their body length was measured. In the groups fed with chicken feed mash, chicken pellets or cattle pellets, the mean body lengths were 2.88, 2.92 and 2.95 cm, respectively, similar to the 2.95 cm of the control T. molitor fed with wheat bran. Individual weight measurements yielded similar results: control, 7.87 g; chicken feed mash, 7.43 g; chicken pellets, 7.60 g; and cattle pellets, 7.63 g. Analysis of livestock feed intake showed that the intake was 290.4 g in control insects, 137.7 g in insects fed chicken feed mash, 220.2 g in insects fed chicken pellets and 230.3 g in insects fed cattle pellets. In the excretion analysis, the excretion of the control insects (161.60 g) was larger than that of the insects fed with chicken feed mash (93.37 g), chicken pellets (93.40 g) or cattle pellets (133.87 g). An analysis of the nutritional content of T. molitor based on nine major nutrients showed no significant differences between the control and the test groups. These results can be used to improve the price competitiveness and quality of T. molitor for commercial development. [ABSTRACT FROM AUTHOR]

The occurrence of antibiotic-resistant bacteria in foodstuff involves a human health risk. Edible insects are a precious resource; however, their consumption raises food safety issues. In this study, the occurrence of antibiotic resistant bacteria in laboratory-reared fresh mealworm larvae (Tenebrio molitor L.) and frass was assessed. Antibiotics were not used during the rearing. Enterobacteriaceae and enterococci were isolated from 17 larvae and eight frass samples. In total, 62 and 69 isolates presumed to belong to Enterobacteriaceae and Enterococcus spp., respectively, were obtained and tested for antibiotic susceptibility via disk diffusion. Based on the results, isolates were grouped, and representative resistant isolates were identified at species level through 16S rRNA gene sequencing. For enterococci resistance, percentages higher than 15% were observed for vancomycin and quinupristin–dalfopristin, whereas Enterobacteriaceae resistance higher than 25% was found against cefoxitin, ampicillin, and amoxicillin–clavulanic acid. Based on the species identification, the observed resistances seemed to be intrinsic both for enterococci and Enterobacteriaceae, except for some β-lactams resistance in Shigella boydii (cefoxitin and aztreonam). These could be due to transferable genetic elements. This study suggests the need for further investigations to clarify the role of edible insects in the spreading of antibiotic resistance determinants through the food chain. [ABSTRACT FROM AUTHOR]

Methanolic stem bark extracts from ten Mexican Bursera Jacq. ex L. species were evaluated in vitro with regard to their inhibitory activity against two Tenebrio molitor-derived enzymes. Seven extracts (B. bicolor, B. copallifera, B. fagaroides, B. grandifolia, B. lancifolia, B. linanoe, and B. longipes) reduced α-amylase activity by 55.37% to 96.25%, with three samples proving to be particularly potent α-amylase inhibitors (B. grandifolia, B. lancifolia, and B. linanoe; IC50 = 162, 132, and 186 µg/mL, respectively). In contrast, no extract inhibited acetylcholinesterase activity by more than 39.94%. Quantitative HPLC analysis did not reveal any clear correlation between the species-specific flavonoid or phenolic acid profiles and the respective extracts' enzyme inhibitory activity. The findings reported herein do not only contribute to improving the current state of knowledge regarding the enzyme inhibitory potential of the Bursera genus, but could also lead to the development of new sustainable bioinsecticides. [ABSTRACT FROM AUTHOR]

Background: This study aimed to evaluate the effects of Hermetia illucens (Black soldier fly-BSF) and Tenebrio molitor (Yellow mealworm-YMW) live larvae as a new nutritional model on duck's gut health, considering gut histomorphometry, mucin composition, cytokines transcription levels, and microbiota. A total of 126, 3-days-old, females Muscovy ducks were randomly allotted to three dietary treatments (6 replicates/treatment, 7 birds/pen): (i) C: basal diet; (ii) BSF: C + BSF live larvae; (iii) YMW: C + YMW live larvae. BSF and YMW live larvae were administered on top of the basal diet, based on the 5% of the expected daily feed intake. The live weight, average daily gain, average daily feed intake and feed conversion ratio were evaluated for the whole experimental period. On day 52, 12 ducks/treatment (2 birds/replicate) were slaughtered and samples of duodenum, jejunum, ileum, spleen, liver, thymus and bursa of Fabricius were collected for histomorphometry. Mucin composition was evaluated in the small intestine through histochemical staining while jejunal MUC-2 and cytokines transcription levels were evaluated by rt-qPCR. Cecal microbiota was also analyzed by means of 16 S rRNA gene sequencing. Results: Birds' growth performance and histomorphometry were not influenced by diet, with a proximo-distal decreasing gradient from duodenum to ileum (p < 0.001), respecting the physiological gut development. Mucin staining intensity and MUC-2 gene expression did not vary among dietary treatments, even though mucin intensity increased from duodenum to ileum, according to normal gut mucus physiology (p < 0.001). Regarding local immune response, IL-6 was higher in YMW group when compared to the other groups (p = 0.009). Insect live larvae did not affect cecal microbiota diversity, but BSF and YMW groups showed a higher presence of Helicobacter, Elusimicrobium, and Succinatimonas and a lower abundance of Coriobacteriaceae and Phascolarctobacterium compared to C birds (p < 0.05). Conclusions: The use of BSF and YMW live larvae as new nutritional model did not impair gut development and mucin composition of Muscovy ducks, but slightly improved the intestinal immune status and the microbiota composition by enhancing regulatory cytokine IL-6 and by increasing minor Operational Taxonomic Units (OTUs) involved in short-chain fatty acids production. [ABSTRACT FROM AUTHOR]

This study investigates the encapsulation of Tenebrio molitor hydrolysate exhibiting DPP-IV inhibitory activity by spray-drying and electrospraying techniques. First, we optimized the feed formulation and processing conditions required to obtain nano-microcapsules by electrospraying when using Arabic gum as an encapsulating agent and pullulan and Tween 20 as additives. The optimum formulation was also dried by spray-drying, where the removal of the additives was also assayed. Morphology analysis reveals that electrosprayed capsules have a smaller size (1.2 ± 0.5 µm vs. 12.4 ± 8.7 µm) and greater uniformity compared to those obtained by spray-drying. Regarding the surface nitrogen content and DPP-IV inhibitory activity, our results show no significant difference between the electrosprayed capsules and spray-dried capsules containing additives (IC50 of ~1.5 mg protein/mL). Therefore, it was concluded that adding additives during spray-drying allows for a similar encapsulation efficiency and reduced degradation during processing, as achieved by electrospraying technique but providing higher productivity. On the other hand, spray-dried capsules without additives displayed a higher surface nitrogen content percentage, which was mainly due to the absence of Tween 20 in the feed formulation. Consequently, these capsules presented a higher IC50 value (IC50 of 1.99 ± 0.03 mg protein/mL) due to the potential degradation of surface-exposed peptides. [ABSTRACT FROM AUTHOR]

Non‐linear immune response is one of the causes of multi‐stability of infection outcomes. In order to avoid the uncertainty caused by multi‐stability in experiments, one needs to quantitatively measure immune response and other traits as well. Here, we took a quantitative approach, which combines mathematical model and time‐series data, to measure the immune response of the Tenebrio moliter challenged by non‐replicative immune inducers. The results showed that the host immunity that characterized by in‐vitro bacterial killing rapidly mounted after immune challenging, then slowly declined to an undetectable level within 100 h. We then quantified the immunity at 15 h after challenging. The results showed that immune response was non‐linearly correlated with the titter of inducer, which fits well to a Hill‐like function. Our results have verified the non‐linear immune response with respect to amount of invading pathogens and may also help to rigorously test the concepts and theories in host–pathogen interaction, such as resistance and tolerance. [ABSTRACT FROM AUTHOR]

Polystyrene (PS) is an extremely stable polymer with a relatively high molecular weight and a strong hydrophobic character that makes it highly resistant to biodegradation. In this study, PS was subjected to biodegradation tests by Tenebrio Molitor (T. Molitor) and Zophobas Morio (Z. Morio) larvae. Specifically, six different experimental diets were compared: (i) T. Molitor fed with bran; (ii) T. Molitor fed only PS; (iii) T. Molitor fed only PS treated with H2O2; (iv) Z. Morio fed with bran; (v) Z. Morio fed only PS; and (vi) Z. Morio fed only PS treated with H2O2. Therefore, the mass change of the larvae and the survival rate were measured periodically, while the frass collected after 15 and 30 days was analyzed by different analyses, such as spectroscopy (FTIR), spectrometry (molecular weight and polydispersity), thermal analysis (TGA) and microscopy (scanning electron microscopy observations). The obtained results suggest that in the case of T. Molitor larvae, larvae feeding on bran showed the highest survival rate of ~94% at 30 days, while in the case of the Z. Morio larvae, the highest survival rate was exhibited by larvae eating PS-H2O2. Although not strongly pronounced, the Mw and Mn of PS in the frass of both T. Molitor and Z. Morio larvae decreased over 30 days, suggesting PS biodegradation. Finally, the morphological analysis shows that PS samples isolated from the frass of T. Molitor and Z. Morio larvae showed completely different, rough and irregularly carved surface structures, in comparison to PS before biodegradation. [ABSTRACT FROM AUTHOR]

With the world's population continuing to grow, ensuring sustainable protein sources for everyone is becoming increasingly challenging. Despite meat being considered unsustainable, people find it challenging to abstain from consuming it. However, one solution to this dilemma could be the incorporation of mealworms into conventional meat products, i.e., sausages. The incorporation of mealworms into sausage formulations appears to shift the fatty acid profile towards higher levels of monounsaturated fats and polyunsaturated fatty acids (PUFAs), particularly omega-3s, potentially enhancing the nutritional value and offering health benefits. Therefore, our study aimed to improve the nutritional value and safety parameters of traditional sausages by enriching them with the flour of mealworm larvae. For this purpose, the larvae were reared on a sustainable substrate with brewery by-products, brewer's yeast, and carrots. They were used frozen and freeze-dried in sausage recipes, replacing pork in different proportions. The analysis of the product's chemical safety parameters (biogenic amines, nitrates and nitrites, volatile fatty acids (FA), and peroxide) and nutritional value (including collagen, cholesterol, amino acids, FA, and hydroxyproline) was carried out in an accredited laboratory. The results of our study have demonstrated that the incorporation of mealworms into sausages, particularly through freeze-drying, increased fat content and enhanced the profile of FA, including omega-3s while reducing protein and cholesterol levels, and altering collagen content, suggesting improved nutritional value and potential health benefits without compromising the safety of the product. Therefore, we are highlighting that the addition of mealworms influences the quality of amino acids positively and maintains biogenic amine levels within safe limits, alongside a negligible impact on nitrates and nitrites and a reduction in peroxide values. These findings indicate an overall improvement in sausage quality and safety without compromising safety. [ABSTRACT FROM AUTHOR]

In pursuit of sustainable protein sources, the agricultural sector and emerging edible insect industry intersect in the valorization of agricultural by-products. Establishing a mutually beneficial relationship involves utilizing agricultural by-products as feeding substrates for insect farming, potentially enhancing the sustainability of both sectors. In the present study, by-products from beer, rice, oat, maize, sunflower, and lucerne, as well as mill residues and spent mushroom substrate from the regions of Thessaly and Central Macedonia (Greece) were investigated as nutritional sources for the larvae of the yellow mealworm (Tenebrio molitor). Results show that the suitability of the tested by-products for rearing T. molitor larvae varies greatly, with larvae surviving better in some by-products than in others. The highest survival rate and the highest weight of larvae were recorded for larvae reared on rice bran, spent grains, and oat by-products. Similarly, high feed conversion and growth rate were observed when the larvae were fed with rice bran and spent grains. Thus, this research promotes cost-effective and sustainable T. molitor rearing, aligning with circular economy principles. [ABSTRACT FROM AUTHOR]

The mealworm Tenebrio molitor L. (Coleoptera: Tenebrionidae) feeds on wheat bran and is considered both a pest and an edible insect. Its larvae contain proteins and essential amino acids, fats, and minerals, making them suitable for animal and human consumption. Zearalenone (ZEA) is the mycotoxin most commonly associated with Fusarium spp. It is found in cereals and cereal products, so their consumption is a major risk for mycotoxin contamination. One of the most important effects of ZEA is the induction of oxidative stress, which leads to physiological and behavioral changes. This study deals with the effects of high doses of ZEA (10 and 20 mg/kg) on survival, molting, growth, weight gain, activity of antioxidant enzymes superoxide dismutase (SOD) and glutathione S-transferase (GST), and locomotion of mealworm larvae. Both doses of ZEA were found to (i) have no effect on survival, (ii) increase molting frequency, SOD, and GST activity, and (iii) decrease body weight and locomotion, with more pronounced changes at 20 mg/kg. These results indicated the susceptibility of T. molitor larvae to high doses of ZEA in feed. [ABSTRACT FROM AUTHOR]

Frass, a byproduct of insect rearing, has become popular for its potential use in sustainable agriculture. The rapid growth of insect production results in an increased frass output. This study examined the effects of frass as soil amendment on plant growth and resistance to insect herbivory. In greenhouse experiments, Brassica rapa L. (Brassicaceae), was grown in unamended soil (NoFrass; control) or soil amended with frass (2 g kg−1) from larvae of black soldier fly (BSFF), Hermetia illucens L. (Diptera: Stratiomyidae), or yellow mealworm (MWF), Tenebrio molitor L. (Coleoptera: Tenebrionidae). Frass was applied as raw, incubated, or composted frass before seed germination. Plant growth and performance were measured of larvae of root‐feeding Delia radicum L. (Diptera: Anthomyiidae) and shoot‐feeding Plutella xylostella L. (Lepidoptera: Plutellidae). Initially, raw BSFF and MWF reduced the growth of B. rapa and resulted in a smaller leaf area than NoFrass. However, over time, a notable trend emerged. Whereas the difference in leaf area between MWF and NoFrass disappeared, BSFF consistently resulted in a smaller leaf area than MWF and NoFrass. Raw BSFF reduced D. radicum larval survival and pupal biomass and larval survival of P. xylostella. In contrast, raw MWF increased larval survival and biomass of D. radicum and the survival of P. xylostella larvae. Interestingly, incubation of frass in the soil for 16 days before seed germination removed plant growth inhibition and increased plant leaf area, especially for MWF compared to NoFrass. In addition, composting MWF increased leaf growth. Therefore, frass may be used as a sustainable and natural alternative to conventional organic fertilisers, promoting plant growth and enhancing resistance to herbivory. Our results indicate that soil amendment with raw BSFF may negatively impact herbivore performance, whereas raw MWF may enhance herbivore performance. [ABSTRACT FROM AUTHOR]

Simple Summary: Mealworms are edible insects that are used as food ingredients because nearly all of their edible sections are lipid and protein sources that humans can consume. Triacylglycerols are significant energy sources and the main components of mealworm oil. Due to the composition of triacylglycerol being related in basic properties to oils, the determination of the triacylglycerol composition is important to understand oils. Our study used electrospray ionization tandem mass spectrometry with multiple neutral loss scans to analyze the content and composition of triacylglycerol in mealworm oil. The acquired scan data were deconvoluted to minimize isotopic interference. Additionally, the validation parameters showed relatively good results. The mealworm oil mainly contained triacylglycerol possessing palmitic acid, oleic acid and linoleic acid. The influence of triacylglycerol on mealworm oil is relatively stronger against oxidative stress compared to other vegetable oils. Mealworms (Tenebrio molitor) have been used as an alternative source of proteins and lipids. Triacylglycerols (TAGs) are major sources of energy and have been used to provide essential fatty acids. They are also the main components of mealworm oil, and their composition and content are extensively linked to its physical and chemical properties. However, because of the complexity of TAG molecules, their identification and quantitation are challenging. This study employed electrospray ionization tandem mass spectrometry (ESI-MS/MS) with multiple neutral loss scans (NLS) to analyze the TAG composition and content in mealworm oil. Identifying and quantifying TAGs using ESI-MS/MS in combination with multiple NLS was an efficient way to improve accuracy and timeliness. For the accurate quantification of TAGs, isotopic deconvolution and correlation factors were applied. A total of 57 TAGs were identified and quantified: C52:2 (16:0/18:1/18:1) (1549.4 nmol/g, 18.20%), C52:3 (16:0/18:1/18:2) (1488.1 nmol/g, 17.48%), C54:4 (18:1/18:1/18:2) (870.1 nmol/g, 10.23%), C54:6 (18:1/18:2/18:2) (659.8 nmol/g, 7.76%) and C52:4 (16:0/18:2/18:2) (600.5 nmol/g, 7.06%), which were the most abundant TAGs present in the mealworm oil. The fundamental properties of mealworm oil, including its degree of oxidation, nutritional effect and physical properties, were elucidated. [ABSTRACT FROM AUTHOR]