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Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on frozen and dried formulations from whole lesser mealworm (Alphitobius diaperinus larva) as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The term lesser mealworm refers to the larval form of the insect species Alphitobius diaperinus. The NF comprises the frozen and freeze‐dried formulations of the lesser mealworm as whole or in the form of a paste or powder. Apart from water in the frozen formulations (whole, paste), the main components of the NF are crude protein and fat, besides smaller amounts of digestible carbohydrates and fibre (chitin). The Panel notes that the levels of contaminants in the NF depend on the concentration of such substances in the insect feed. The Panel notes furthermore that the true protein levels in the NF are overestimated when using the nitrogen‐to‐protein conversion factor of 6.25, due to the presence of non‐protein nitrogen from chitin. The applicant proposed to use the NF formulations added as an ingredient to various food products such as cereal bars, pasta, meat imitates and bakery products. The target population is the general population. Additionally, the applicant proposed to use the NF as a food supplement in adults. The Panel notes that, considering that the NF will not be the sole source of dietary protein, and the composition of the NF and the proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous. The submitted subchronic 90‐day toxicity study with the NF as testing material did not raise safety concerns. The Panel considers that the consumption of the NF may induce primary sensitisation and allergic reactions to lesser mealworm proteins and may cause allergic reactions in subjects with allergy to crustaceans and dust mites. Additionally, allergens from the feed may end up in the NF. Allergenicity aside, the Panel concludes that the NF is safe under the proposed uses and use levels.

In this study, we aimed to obtain hydrolysates with bioactive peptides from mealworm (Alphitobius diaperinus L.) larvae using an artichoke (Cynara scolymus L.) enzyme extract. Two types of substrates were used: the raw larvae flour (LF) and its protein extract (PE). The hydrolysis yield, considering the peptide concentration of the hydrolysates, was higher in PE hydrolysates than in LF hydrolysates (6.39 ± 0.59 vs. 3.02 ± 0.06 mg/mL, respectively). However, LF showed a higher antioxidant activity against the DPPH radical than PE (59.10 ± 1.42 vs. 18.79 ± 0.81 µM Trolox Eq/mg peptides, respectively). Regarding the inhibitory activity of angiotensin-I-converting enzyme (ACE), an IC50 value of 111.33 ± 21.3 µg peptides/mL was observed in the PE. The identification of the peptide sequence of both hydrolysates was conducted, and LF and its PE presented 404 and 116 peptides, respectively, most with low molecular weight (<3 kDa), high percentage of hydrophobic amino acids, and typical characteristics of well-known antioxidant and ACE-inhibitory peptides. Furthermore, the potential bioactivity of the sequences identified was searched in the BIOPEP database. Considering the antioxidant and ACE-inhibitory activities, LF hydrolysates contained a larger number of sequences with potential bioactivity than PE hydrolysates.

The effect of spinosad exposure on the susceptibility of pyrethroid- and organophosphate-resistant populations of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), to insecticides was investigated in broiler house farm and laboratory studies. A field pyrethroid- and organophosphate-resistant population showed a 3.6-fold increase in susceptibility to gamma -cyhalothrin following spinosad treatment. Overall, cyfluthrin- and fenitrothion-resistant field populations were more susceptible to these insecticides following spinosad treatments, but populations that were not resistant showed no change in susceptibility following spinosad treatment. In a related study, three broiler farm beetle populations with very similar levels of cyfluthrin and gamma -cyhalothrin resistance and similar susceptibilities to spinosad were used to investigate temporal effects of spinosad field treatments on the susceptibility to pyrethroids. Farm insecticide regimes applied at the start of each flock differed: the control broiler house received no insecticide applications, another house was systematically treated with cyfluthrin at the start of each study flock, and the third house was systematically treated with spinosad at the start of five flocks. Afterwards, treatments reverted to cyfluthrin on all farms. At the end of flocks, beetles were tested with cyfluthrin, gamma -cyhalothrin, and spinosad. The control and cyfluthrin house beetles did not change susceptibility to pyrethroids over the period of the study. In the spinosad house, spinosad had no effect on spinosad susceptibility but dramatically increased cyfluthrin and gamma -cyhalothrin susceptibilities. These new susceptibilities were maintained while spinosad applications continued, but pyrethroid susceptibility declined once spinosad applications ceased. This study provides evidence of a synergistic interaction between spinosad and pyrethroids in pyrethroid-resistant beetles. This evidence has significant implications for management of insecticide-resistant populations through an integrated spinosad-pyrethroid strategy that aims to minimize insecticide use while enhancing control.

In response to growing food demand, edible insects are perceived as an opportunity to alleviate food insecurity. With its wide edible insects’ biodiversity, the Democratic Republic of Congo is one of Africa’s most critical entomophagous. This study aimed at giving a first insight on inventory showing diversity, perception, consumption, availability, host plants, harvesting techniques and processing techniques of edible insects in South-Kivu, DRC. It recorded twenty-three edible insects belonging to nine families and five orders, some of which are consumed in the larval, adult, egg and pupa stages. Rhyncophorus phoenicis , Alphitobius diaperinus , Macrotermes subhyalinus and Acheta domesticus were the most preferred edible insects in Fizi Territory, Ruspolia differens and Apis mellifera larvae in Kabare Territory, Imbrasia oyemensis , Imbrasia epimethea , Rhynchophorus ferrugineus and Rhyncophorus phoenicis in Mwenga Territory, Ruspolia differens , Macrotermes subhyalinus , Gryllotalpa africana , Nsike, Nomadacris septemfasciata and A. mellifera larvae in Walungu Territory. Ruspolia differens , I. oyemensis , A. mellifera larvae, G. africana and Nsike, were preferred for their taste. Acheta domesticus , A. diaperinus and A. mellifera larvae were abundant throughout the year, while others were only available for 9 months or less per year. Numerous plants have been recorded as their hosts, including plants used for food and income. Harvesting strategies and period, processing methods and preservation techniques depend on insect species, local knowledge and practices. These findings suggest similar and thorough studies on entomophagy across the country while encouraging the rearing of edible insects to address their existing high demand and environmental concerns.

This study aimed to investigate the potential accumulation of mycotoxins in the lesser mealworm (Alphitobius diaperinus, LMW) and black soldier fly (Hermetia illucens, BSF) larvae. Feed was spiked with aflatoxin B1, deoxynivalenol (DON), ochratoxin A or zearalenone, and as a mixture of mycotoxins, to concentrations of 1, 10, and 25 times the maximum limits set by the European Commission for complete feed. This maximum limit is 0.02 mg/kg for aflatoxin B1, 5 mg/kg for DON, 0.5 mg/kg for zearalenone and 0.1 mg/kg for ochratoxin A. The mycotoxins and some of their metabolites were analysed in the larvae and residual material using a validated and accredited LC-MS/MS-based method. Metabolites considered were aflatoxicol, aflatoxin P1, aflatoxin Q1, and aflatoxin M1, 3-acetyl-DON, 15-acetyl-DON and DON-3-glycoside, and α- and β-zearalenol. No differences were observed between larvae reared on mycotoxins individually or as a mixture with regards to both larvae development and mycotoxin accumulation/excretion. None of the mycotoxins accumulated in the larvae and were only detected in BSF larvae several orders of magnitude lower than the concentration in feed. Mass balance calculations showed that BSF and LMW larvae metabolized the four mycotoxins to different extents. Metabolites accounted for minimal amounts of the mass balance, except for zearalenone metabolites in the BSF treatments, which accounted for an average maximum of 86% of the overall mass balance. Both insect species showed to excrete or metabolize the four mycotoxins present in their feed. Hence, safe limits for these mycotoxins in substrates to be used for these two insect species possibly could be higher than for production animals. However, additional analytical and toxicological research to fully understand the safe limits of mycotoxins in insect feed, and thus the safety of the insects, is required.

New liquid fermentation techniques for the production of the bioinsecticidal fungus Metarhizium brunneum strain F-52 have resulted in the formation of microsclerotia (MS), a compact, melonized-hyphal structure capable of surviving desiccation and formulation as dry granules. When rehydrated, these MS granules germinate to produce conidia that can infect susceptible insects. Fermentation media containing cottonseed or soy flours as nitrogen sources and formulated at two carbon to nitrogen ratios (C:N), 30:1 or 50:1, were evaluated for production of microsclerotia. Dry MS granule samples were compared for storage stability based on conidia production, and insecticidal activity against larvae of the lesser mealworm, Alphitobius diaperinus (Panzer), using a potting soil bioassay. Cottonseed and soy flours were equivalent for production, MS granule viability, and insecticidal activity. Fermentation media containing higher nitrogen concentrations (30:1 C:N) resulted in greater biomass accumulation and greater production of conidia from granules regardless of the nitrogen source. MS granules made with M. brunneum cultures grown in media with 30:1 C:N produced 8.5 109 conidia per gram of granules after 8-d incubation, significantly higher than MS granules made using fungus produced using 50:1 C:N media (5.5 109 conidia per gram dry MS granules). The LC50 for larval mortality was 8.05 105 conidia per cup, equivalent to applications of 94 or 147 mu g granules per cup for granules made from high and low nitrogen media, respectively. Measurements of water activity were not significantly different among granule samples (0.28-0.29) even though granules made from high nitrogen media had higher moisture content (>5.2%) compared with granules made from low nitrogen media (<4.6%). Higher initial conidial production was reflected in longer storage stability at 25 degree C, with half-lives estimated at 3.7 and 1.7 wk for 30:1 and 50:1 C:N ratios, respectively. These results support further evaluation of MS granule formulations for the control of soil-inhabiting insect pests.

One of the general trends in the food industry is the production of new healthy snack products. Extrusion is one of the most versatile and efficient food processing techniques used in preparation of starch-based foods. Alternative sources of nutrients such as proteins and minerals are interesting ingredients to add to these products. The aim of this work was to evaluate the effects of enrichment with Alphitobius diaperinus, Tenebrio molitor, and pea protein powder on physicochemical and nutritional properties of extruded snacks. Extrudates were made using a single-screw extruder, with 5% of enrichment materials and 95% corn grits. Water content, water activity, particle size, expansion index, bulk density, porosity, water absorption index, water solubility index, swelling index, hygroscopicity, colour, texture, protein, and mineral content were assessed. The results show that enrichment ingredients are a practical alternative in extrudate formulation, maintaining physicochemical characteristics of the products and improving nutritional aspects.

Insects are a promising alternative protein source. One of the bottlenecks in applying insects in food is the fast darkening initiated during grinding. Besides enzymatic browning, non-enzymatic factors can cause off-colour formation, which differs between species. This study investigates the impact of iron, phenoloxidase, and polyphenols on off-colour formation in insect larvae. Hermetia illucens showed a blackish colour, whereas Tenebrio molitor turned brown and Alphitobius diaperinus remained the lightest. This off-colour formation appeared correlated with the iron content in the larvae, which was 61 ± 9.71, 54 ± 1.72 and 221 ± 6.07 mg/kg dw for T . molitor , A . diaperinus and H . illucens , respectively. In model systems, the formation of iron-L-3,4-dihydroxyphenylalanine (L-DOPA) bis- and tris-complexes were evidenced by direct injection into ESI-TOF-MS, based on their charges combined with iron isotope patterns. The reversibility of the binding of iron to phenolics, and thereby loss of blackening, was confirmed by EDTA addition. Besides complex formation, oxidation of L-DOPA by redox reactions with iron occurred mainly at low pH, whereas auto-oxidation of L-DOPA mainly occurred at pH 10. Tyrosinase (i.e. phenoloxidase) activity did not change complex formation. The similarity in off-colour formation between the model system and insects indicated an important role for iron-phenolic complexation in blackening.

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.HighlightsSuperworm larvae showed the highest content of SFAMealworms larvae were the richest in PUFAThe fat-soluble vitamin content followed the fatty acids content

Edible insect powder is a new ingredient that can be used to create innovative functional and nutritional properties in food products. However, adding insect powder to bakery goods could impact the dough’s technological and sensory attributes. This study aims to investigate possible alterations in dough enriched with insect powder and examine the texture of salty snacks produced using a 3D food printer. The doughs were prepared using varying quantities of two types of edible insect powders. The stiff flow properties showed a pseudoplastic behaviour that fit well with a power-law equation. Viscosity and pseudoplasticity increased in proportion to insect powder concentration, with the strongest effect observed for L. migratoria. The linear viscoelastic behaviour exhibited G′ values exceeding G″ values, and both moduli increased with insect concentration. A. migratoria displayed a greater elastic contribution (lower tan δ) compared to A. diaperinus, which correlates with the lower printability of A. migratoria. The texture of the snacks prepared with A. diaperinus demonstrated a rise in the breaking force as the insect concentration increased and exhibited a corresponding texture profile to the control and the 4.6% sample. Moreover, the addition of A. diaperinus heightened the crispiness.