Tenebrio molitor as a source of interesting natural compounds, their recovery processes, biological effects, and safety aspects.

Nowadays, it is urgent to produce in larger quantities and more sustainably to reduce the gap between food supply and demand. In a circular bioeconomy vision, insects receive great attention as a sustainable alternative to satisfy food and nutritional needs. Among all insects, Tenebrio molitor (TM) is the first insect approved by the European Food Safety Authority as a novel food in specific conditions and uses, testifying its growing relevance and potential. This review holistically presents the possible role of TM in the sustainable and circular solution to the growing needs for food and nutrients. We analyze all high value-added products obtained from TM (powders and extracts, oils and fatty acids, proteins and peptides, and chitin and chitosan), their recovery processes (evaluating the best ones in technical and environmental terms), their nutritional and economical values, and their biological effects. Safety aspects are also mentioned. TM potential is undoubted, but some aspects still need to be discussed, including the health effects of substances and microorganisms in its body, the optimal production conditions (that affect product quality and safety), and TM capacity to convert by-products into new products. Environmental, economic, social, and market feasibility studies are also required to analyze the new value chains. Finally, to unlock the enormous potential of edible insects as a source of nutritious and sustainable food, it will be necessary to overcome the cultural, psychological, and regulatory barriers still present in Western countries.

Sensory and Chemical Characteristic of Two Insect Species: Tenebrio molitor and Zophobas morio Larvae Affected by Roasting Processes.

The volatile compounds from insects (Tenebrio molitor and Zophobas morio larvae) roasted at 160, 180, or 200 °C and fed with potato starch or blue corn flour were isolated by solid-phase microextraction (SPME), and identified by gas chromatography-mass spectrometry (GC-MS). In the tested material, 48 volatile compounds were determined. Among them, eight are pyrazines, aroma compounds that are formed in food products during thermal processing due to the Maillard reaction. Eleven of the identified compounds influenced the roast, bread, fat, and burnt aromas that are characteristic for traditional baked dishes (meat, potatoes, bread). Most of them are carbonyl compounds and pyrazines. To confirm the contribution of the most important odorants identified, their odor potential activity values (OAVs) and %OAV were calculated. The highest value was noted for isobuthylpyrazine, responsible for roast aroma (%OAV > 90% for samples roasted at lower temperatures), and 2,5-dimethylpyrazine, responsible for burnt aroma (%OAV > 20% for samples roasted at the highest temperature). According to the study, the type of feed did not significantly affect the results of the sensory analysis of roasted insects. The decisive influence was the roasting temperature. The highest scores were achieved for Tenebrio molitor larvae heat-treated at 160 °C.

Utilization of insect proteins to formulate nutraceutical delivery systems: Encapsulation and release of curcumin using mealworm protein-chitosan nano-complexes.

The potential of using insect proteins to encapsulate and protect hydrophobic nutraceuticals within biopolymer nano-complexes was examined. Insect proteins were used to form nanoparticles that were uncoated or coated with chitosan. Initially, the nature of the curcumin-mealworm protein interaction was investigated. Curcumin mainly interacted with the hydrophobic core of the insect protein nanoparticles through hydrophobic forces. About one curcumin molecule bound per protein molecule in both the absence and presence of chitosan. The binding constants (K) were 1.1 × 104 M-1 and 0.7 × 104 M-1 for curcumin loaded in the uncoated and coated nanoparticles, respectively. Differential scanning calorimetry showed increased thermal stability of the proteins after interaction with curcumin or chitosan. Encapsulation efficiency of the curcumin within the biopolymer nano-complexes was 30-47% depending on the system. Transmission electron microscopy and dynamic light scattering analysis showed that the biopolymer nano-complexes were spherical and relatively small (d = 143-178 nm). FTIR suggests that curcumin was stabilized more effectively in the coated nano-complexes, due to non-covalent intermolecular interactions. Curcumin release under oral, gastric, and intestinal conditions showed that over 90% of the nutraceutical was released after exposure to model gastrointestinal conditions. The findings demonstrate the potential of using insect proteins for fabricating colloidal delivery systems for water-insoluble nutraceuticals.

The effect of fish and mealworm larvae meals as alternative dietary protein sources on nutrient digestibility and gastrointestinal function in Chinchilla lanigera.

Chinchillas are herbivores, but wild chinchillas may occasionally consume animal-based foods. The aim of this study was to determine the effect of fish meal (FM) and mealworm meal (MWM) included in complete pelleted diets on nutrient digestibility and gastrointestinal function in chinchillas. The experiment was performed on 24 male, divided into three groups, n=8. Control group (C) was fed a diet containing 10% soybean meal (SBM). In the experimental group FM, chinchillas received a diet containing 3% fish meal, and the diet administered to the experimental group MWM was supplemented with 4% dried mealworm larvae meal. The nutrient digestibility of diets was determined. At the end of the experiment animals were euthanized and their digestive tracts were removed to analyze gut activity. FM group animals were characterized by lower crude fat digestibility, whereas both alternative protein sources improved the digestibility of acid detergent fiber (ADF). A considerable increase in the activity of cecal intracellular and extracellular bacterial enzymes (in particular ß-glucosidase, ß-galactosidase and ß-xylosidase) was noted in the FM group, which however did not increase the concentrations of short-chain fatty acids (SCFA). The inclusion of MWM in chinchilla diets shifted the bacterial fermentation site from the cecum (lowest SCFA pool) to the colon (highest SCFA pool), thus enabling to derive additional energy from less digestible dietary components. In conclusion, chinchilla diets can be supplemented with small amounts of animal protein such as fish meal and dried mealworm larvae meal.

Influence of Feeding Substrates on the Presence of Toxic Metals (Cd, Pb, Ni, As, Hg) in Larvae of Tenebrio molitor: Risk Assessment for Human Consumption.

Larvae of Tenebrio molitor L. (Coleoptera: Tenebrionidae) are particularly suitable as novel food for the human consumption; nevertheless, there are some potential safety risks linked with insect consumption. In this study we investigated the presence of Cd, Pb, Ni, As, Hg in new feeding substrates coming from solid residues generated by olive fruits processing, called olive-pomace, and their influence on the metal content in larvae of T. molitor. Morover, bioaccumulation factor and the mercury-selenium balance were evaluated. Analyses were carried out via graphite furnace atomic absorption spectrophotometry for Cd, Pb, Ni, As and Se, and via Direct Mercury Analyzer for Hg. All metal concentrations found in feeding substrates were below the legal limit of undesirable substances in animal feed (2002/32/EC). Concentrations in larvae were in the range (mg kg-1 wet weight): Cd 0.008-0.016, Pb 0.063-0.079, Ni 0.03-0.63, As 0.021-0.023, Hg 0.12 × 10-3-0.49 × 10-3, and Se 0.057-0.085. Statistically significant correlation between metal content in feeding substrates and in larvae was evidenced only for Hg, which bioaccumulates. Se protects from mercury toxicity, with a Selenium Health Benefit Value (HVBSe) of > 0. Overall, our results indicate that the risk of exposure to metals from consumption of mealworm larvae is relatively low and in compliance with European Union regulations.

Chronic Oral Administration of Tenebrio molitor Extract Exhibits Inhibitory Effect on Glucocorticoid Receptor Overexpression in the Hippocampus of Ovariectomy-Induced Estrogen Deficient Mice.

It has been reported that estrogen deficiency in female disrupts systemic endocrinologic regulatory mechanisms, finally leading to osteoporotic condition. Estrogen deficiency also down-regulates brain functions due to its deficits of its original roles in a number of neurological events. Therefore, it is necessary to find alternative materials that can prevent osteoporotic condition and maintain normal brain functions to correct such hormone deficiency. In the present study, we found that novel compounds originated from larvae of Tenebrio molitor (TM) possessed anti-osteoporotic effect. They could also prevent abnormal progressive brain function by deaccelerating enhanced HPA-axis negative feedback while maintaining neurogenesis in hippocampus. We daily administered TM to ovariectomized (OVX) ddY mice for 4 weeks and then performed histological and hormonal evaluations for its anti-osteoporotic effects. In addition, we investigated glucocorticoid receptor (GR) expression and neuroblast expression (DCX) in the hippocampal dentate gyrus morphologically by immunohistochemistry analysis. According to our results, TM has anti-osteoporotic effects. It also tends to bring interfered brain environment back to normal condition. These results suggest that TM might have anti-osteoporosis effect and enhancing effects on enrichment of environment in brain by being antidestroyed hormonal deficiency simultaneously. This is the first study to report that TM can be used as source of bioactive substance to prevent decreased neurogenesis and impaired HPA axis driven by high GR expression in the hippocampus in hormonal deficient female animals. PRACTICAL APPLICATION: Anti-osteoporosis effect and stress resistance due to improved brain function caused by the ingestion of Tenebrio molitor extract were observed in postmenopausal women. T. molitor is available as a nutritional supplement for bone and brain health, which menopausal women need most.

Effects of Insect Protein Supplementation during Resistance Training on Changes in Muscle Mass and Strength in Young Men.

During prolonged resistance training, protein supplementation is known to promote morphological changes; however, no previous training studies have tested the effect of insect protein isolate in a human trial. The aim of this study was to investigate the potential effect of insect protein as a dietary supplement to increase muscle hypertrophy and strength gains during prolonged resistance training in young men. Eighteen healthy young men performed resistance training four day/week for eight weeks. Subjects were block randomized into two groups consuming either an insect protein isolate or isocaloric carbohydrate supplementation within 1 h after training and pre-sleep on training days. Strength and body composition were measured before and after intervention to detect adaptions to the resistance training. Three-day weighed dietary records were completed before and during intervention. Fat- and bone- free mass (FBFM) improved significantly in both groups (Mean (95% confidence interval (CI))), control group (Con): (2.5 kg (1.5, 3.5) p < 0.01), protein group (Pro): (2.7 kg (1.6, 3.8) p < 0.01) from pre- to post-. Leg and bench press one repetition maximum (1 RM) improved by Con: (42.0 kg (32.0, 52.0) p < 0.01) and (13.8 kg (10.3, 17.2) p < 0.01), Pro: (36.6 kg (27.3, 45.8) p < 0.01) and (8.1 kg (4.5, 11.8) p < 0.01), respectively. No significant differences in body composition and muscle strength improvements were found between groups. In young healthy men, insect protein supplementation did not improve adaptations to eight weeks of resistance training in comparison to carbohydrate supplementation. A high habitual protein intake in both Con and Pro may partly explain our observation of no superior effect of insect protein supplementation.

On printability, quality and nutritional properties of 3D printed cereal based snacks enriched with edible insects.

3D printing technology was employed to obtain snacks with a designed cylindrical geometry from wheat flour dough enriched by ground larvae of Yellow mealworms (Tenebrio molitor) as novel source of proteins. The main microstructural features, overall quality, and nutritional attributes were studied as a function of formulation, time and temperature of baking. The addition of ground insects up to 20 g/100 g (d.b.) resulted in softer dough. This caused an overflow in dough deposition producing the increase in diameter, height and weight of snacks. Baking conditions did not alter the overall aspect of the snacks, but modification of the main dimensional and microstructure attributes were observed due to the better water evaporation. The optimization of baking conditions found that 22 min and 200 °C allowed obtaining a maximum desirability of 0.693. Baked in these conditions, the printed snacks enriched with 10 and 20% of ground insects significantly increased the total essential amino acid, from 32.5 (0% insects) to 38.2 and 41.3 g/100 g protein, respectively. The protein digestibility corrected amino acid score increased from 41.6 to 65.2 from 0 to 20% insect enrichment, with lysine and methionine + cysteine being the respective limiting amino acid. Our results evidenced the rational promotion of insects based on nutritional arguments and validated the use of 3D printing as technology to manufacture innovative printed snacks without adverse impact on technological quality.

Oleic acid and linoleic acid from Tenebrio molitor larvae inhibit BACE1 activity in vitro: molecular docking studies.

In our ongoing research to find therapeutic compounds for Alzheimer's disease (AD) from natural resources, the inhibitory activity of the BACE1 enzyme by Tenebrio molitor larvae and its major compounds were evaluated. The T. molitor larvae extract and its fractions exhibited strong BACE1 suppression. The major components of hexane fraction possessing both high yield and strong BACE1 inhibition were determined by thin layer chromatography, gas chromatography, and nuclear magnetic resonance analysis. A remarkable composition of unsaturated long chain fatty acids, including oleic acid and linoleic acid, were identified. Oleic acid, in particular, noncompetitively attenuated BACE1 activity with a half-maximal inhibitory concentration (IC50) value of 61.31 µM and Ki value of 34.3 µM. Furthermore, the fatty acids were stably interacted with BACE1 at different allosteric sites of the enzyme bound with the OH of CYS319 and the NH3 of TYR320 for oleic acid and with the C=O group of GLN304 for linoleic acid. Here, we first revealed novel pharmacophore features of oleic acids and linoleic acid to BACE1 by in silico docking studies. The present findings would clearly suggest potential guidelines for designing novel BACE1 selective inhibitors.

Comparison of cytotoxic extracts from fruiting bodies, infected insects and cultured mycelia of Cordyceps formosana.

A resazurin method was employed to test and compare cytotoxicity of extracts from fruiting bodies, insects and cultured mycelia of Cordyceps formosana against Chinese hamster ovary (CHO) cells. Results showed that the cultured mycelia had much stronger cytotoxicity than that of the fruiting bodies and infected insects. This suggests that using cultured mycelia to substitute a natural Cordyceps may result in poisoning. A combined method of HPLC-PAD-HRMS and cytotoxic analysis revealed that the most toxic compound (Compound 1) was found mainly in the cultured mycelia and also a small amount in the infected insect body of the Cordyceps, but not in the fruiting body. The second toxic compound (Compound 2) was found in all structures of Cordyceps and in cultured mycelia. Different contents of the toxic compounds resulted in the different cytotoxicity of the extracts. Compound 1 and Compound 2 were prepared with preparative HPLC as yellow and orange powders, respectively. Cytotoxic tests showed that the median lethal dose (LD50) against CHO cells of Compound 1 was 18.3 ± 0.2 and 103.7 ± 5.9 µg/mL for Compound 2. Compound 1 and Compound 2 were identified as rugulosin and skyrin by HRMS, UV and NMR data. The two compounds were never previously isolated from the genera Cordyceps and Hirsutella and their cytotoxicity against CHO cells was also reported for the first time.

Elemental concentration in mealworm beetle (Tenebrio molitor L.) during metamorphosis.

Mealworm beetles have been used in numerous experiments as bioindicators. The aim of our experiment was to study the elemental composition in three larvae, pupae and first and second generation adult stages during their life cycle. We selected 180 larvae from a genetically similar population and put them in three groups, in two boxes (60 larvae in each box). Larvae were fed with mashed potato made of the same quality and quantity of potato powder. Then, we selected 10 individuals from each stage to the elemental analysis, using the ICP-OES method. The following elements were analysed in the studied stages: Ca, Cu, Fe, K, Mg, Mn, Na, P, S, Sr and Zn. The results of principal component analysis demonstrated that based on elemental composition, different stages were separated with each other, but in the cases of the three larvae stages, high overlap was found. The results of the GLM ANOVA showed significant differences between the different stages of metamorphosis-based elemental composition. Our results show that the calcium and magnesium were found in a relatively high concentration, while the iron and zinc may be essential elements during the metamorphosis. Our results also show that in insect, the concentration of sodium was higher than in the pupa which may cause by hemolymph. We also demonstrated that the metamorphosis has an effect on the concentration of elements. Our study shows that in the different stages of insects, there are significant changes in the elemental composition of different stages of insects during their metamorphosis.

Characteristics of maize flour tortilla supplemented with ground Tenebrio molitor larvae.

The larva of the Tenebrio molitor, known as the yellow meal worm, is a plague of wheat and flours. Consumption of the raw insects is not well accepted because of their appearance. The objective of the present work was to grow T. molitor larvae under standard conditions, to analyze the chemical composition of the larvae powder, and to prepare supplemented maize tortillas. Protein and fat contents were performed with standard methods. Tenebrio larvae powder had a 58.4% protein content; this protein was rich in essential amino acids such as phenylalanine, tyrosine, and tryptophan; the found values satisfied those recommended by the Food and Agriculture Organization. Fatty acid composition was determined by GC-MS showing high contents of oleic acid and linoleic acid (19.8 and 8.51%, respectively). A large proportion of unsaturated fatty acids of longer chains was detected. Long-chain fatty acids having two or three double bonds have been claimed as highly beneficial to health. Tortillas supplemented with larvae powder had excellent consumer acceptance, and tortilla protein content increased by 2% as well as the amount of essential amino acids. These results show new ways to consume insects and at the same time increase the nutritional value of the original food products.

A comparison of the nitrogen requirements of the eastern pygmy possum, Cercartetus nanus, on a pollen and on a mealworm diet.

The eastern pygmy possum, Cercartetus nanus, is known to feed both on flower products and on invertebrates. This study compares its ability to meet its nitrogen requirements on pollen and on insect larvae. Captive C. nanus were fed diets in which nitrogen was provided either by Eucalyptus pollen or by the mealworm Tenebrio molitor. The apparent digestibility of the nitrogen from both sources was high, with a mean value of 76% for the pollen and 73% for the mealworms. This was much higher than would have been inferred from the common practice of measuring the percentage of empty pollen grains in fecal samples. The truly digestible maintenance nitrogen requirements of C. nanus on pollen were exceptionally low: 2.6 mg N d-1 compared with 9.5 mg N d-1 on mealworms. The value for pollen is the lowest yet recorded for any mammal. The difference between the requirements of C. nanus on the two diets appeared to be related to the composition of the mealworm and pollen protein. The biological value of the pollen nitrogen was exceptionally high for a plant protein, at 72%, whereas the biological value of the mealworm nitrogen was only 42%. This suggests that the amino acid composition of the pollen corresponded more closely to the requirements of C. nanus than the composition of the mealworm protein did. Pollen is an excellent source of nitrogen for C. nanus, and it should be considered as a potential nitrogen source for other flower-feeding animals.

Calcium balance in Drakensberg crag lizards (Pseudocordylus melanotus melanotus; Cordylidae).

Eight nonreproductive female Drakensberg crag lizards (Pseudocordylus melanotus melanotus) were each fed diets of mealworms and calcium capsules with various calcium levels. Excreta were collected and analyzed for calcium and uric acid content. The amount of calcium in the feces was calculated. The lizards appeared to be able to maintain calcium balance at calcium intakes equivalent to 1.4-5.6% calcium in the dry matter of feed. Calcium balance was maintained by adapting intestinal calcium absorption.