Safety of frozen and dried formulations from migratory locust (Locusta migratoria) as a Novel food pursuant to Regulation (EU) 2015/2283

1.1 Background and Terms of Reference as provided by the requestor

On 28 December 2018, the company Fair Insects BV (A Protix Company) submitted a request to the European Commission in accordance with Article 10 of Regulation (EU) 2015/2283 to authorize placing on the market of whole and ground grasshoppers (Locusta migratoria) as a novel food.

The target population is general population, excluding infants and young children. The applicant has also requested data protection under Article 26 of Regulation (EU) 2015/2283.

In accordance with Article 10(3) of Regulation (EU) 2015/2283, the European Commission asks the European Food Safety Authority to provide a scientific opinion on the safety of whole and ground grasshoppers (Locusta migratoria) as a novel food.

The European Commission asks the European Food Safety Authority to evaluate and inform the Commission as to whether and if so, to what extent, the requirements of Article 26(2)(c) of Regulation (EU) 2015/2283 are fulfilled in elaborating its opinion on whole and ground grasshoppers (Locusta migratoria) regarding the proprietary data for which the applicant is requesting data protection.

In the process of the evaluation of this Novel Food, it became apparent that the Commission should amend the title of the mandate in relation to the terms “whole” and “grasshopper”. The term “grasshopper” is generic and does not exclusively refer to Locusta migratoria, and the term “whole” is not accurate as some of the parts may need to be removed prior to its consumption. On that basis, the Commission amended the title to “Revised request for a scientific opinion on frozen and dried formulations from migratory locust (Locusta migratoria) as a novel food”.

1.2 Interpretation of the Terms of Reference

Given the proposed intended uses and in accordance to Art 5 of the Commission Implementing Regulation (EU) 2017/2469 stating ‘where it cannot be excluded that a novel food intended for a particular group of the population would be also consumed by other groups of the population, the safety data provided shall also cover those groups’, it was clarified that the target population is the general population.

The applicant was requested to provide a revised assessment for the anticipated intake considering all population groups.

2.1 Data

The safety assessment of this NF is based on data supplied in the application and information submitted by the applicant following EFSA's requests for supplementary information. Additional information, which was not included in the application, was retrieved by literature search following a search strategy and standard operating procedure as described by Dibusz and Vejvodova (2020).

Administrative and scientific requirements for NF applications referred to in Article 10 of Regulation (EU) 2015/2283 are listed in the Commission Implementing Regulation (EU) 2017/24691.

A common and structured format on the presentation of NF applications is described in the EFSA guidance on the preparation and presentation of an NF application (EFSA NDA Panel, 2016). As indicated in this guidance, it is the duty of the applicant to provide all of the available (proprietary, confidential and published) scientific data (including both data in favour and not in favour) that are pertinent to the safety of the NF.

This NF application includes a request for protection of proprietary data in accordance with Article 26 of Regulation (EU) 2015/2283. The data requested by the applicant to be protected comprise description of the production process, analytical data on the composition of the NF, analytical data on contaminants in the NF, stability and microbiological status, data on NF sales, intake assessment, protein digestibility and DIAAS, genotoxicity and cytotoxicity study.

2.2 Methodologies

The assessment follows the methodology set out in the EFSA guidance on NF applications (EFSA NDA Panel, 2016) and the principles described in the relevant existing guidance documents from the EFSA Scientific Committee. The legal provisions for the assessment are laid down in Article 11 of Regulation (EU) 2015/2283 and in Article 7 of the Commission Implementing Regulation (EU) 2017/2469.

This assessment concerns only the risks that might be associated with consumption of the NF under the proposed conditions of use, and is not an assessment of the efficacy of the NF with regard to any claimed benefit.

3.1 Introduction

The NF subject of the application is formulations of Locusta migratoria sp. (migratory locust), an insect species that belongs to the family of Acrididae. The NF falls under the category ‘food consisting of, isolated from or produced from animals or their parts’, as described in Article 3 of Regulation (EU) 2015/2283. The NF is produced by farming and processing of L. migratoria and consists mainly of protein, fat and fibre (dry basis). The NF is proposed to be marketed as frozen, dried or in the form of powder. The applicant proposes to use the NF as ingredient in various food products such as breakfast cereals, pasta, bakery products, sauces, meat products and meat imitates. Products with the NF can be consumed by the general population.

3.2 Identity of the NF

The NF consists of frozen, dried and ground formulations of Locusta migratoria sp. (migratory locust). The term ‘migratory locust’ refers to the adult of Locusta migratoria, an insect species that belongs to the Family of Acrididae, Subfamily Locustinae, Genus Locusta. The following scientific synonyms have been described in Global Biodiversity Information Facility – (GBIF Secretariat, 2019), Gryllus (Locusta) migratorius Linnaeus 1758; Pachytylus migratorius, Rehn 1902; Gryllus (Locusta) danicus, Linnaeus 1767; Locusta danica, Ikonnikov 1913; Pachytylus danicus, Doi 1932; Locusta migratoria Danica, Ju 1969; Gryllus (Locusta) cinerascens Fabricius, 1781; Pachytylus cinerascens, Walker 1870.

L. migratoria sp. is currently present in various regions worldwide, including Australia, Asia, Africa and Europe (GBIF Secretariat, 2019). The identity of the insect species has been certified by means of morphological identification by the applicant in collaboration with certified taxonomist in the Netherlands. Due to density-dependent phase polyphenism, L. migratoria exists as two phenotypes, solitary and gregarious. The applicant ensures that adults are reared and harvested during solitary phase by controlling rearing conditions.

The NF is intended to be marketed as A) blanched and frozen L. migratoria (LM frozen); B) blanched and freeze-dried L. migratoria (LM dried); C) blanched, freeze-dried and ground L. migratoria (LM powder). The insects are farmed under controlled rearing conditions.

3.3 Production process

According to the information provided, the NF is produced in line with Hazard Analysis Critical Control Points (HACCP) principles. The applicant stated that insects were reared at a facility registered at the Netherlands Food and Consumer Product Safety Authority (NVWA) as food-producing company. The production process can be divided into three distinct phases, i.e. farming, harvest and post-harvest processing. All steps take place under controlled rearing conditions.

Farming includes mating of the adult insect population and rearing of the nymphs. The eggs are separated from the adult insects so that nymphs can consequently grow separately. After being hatched from the eggs, the nymphs grow under monitored temperature and humidity conditions, in stainless steel containers, certified for food contact and regularly disinfected. The applicant reported that no antimicrobials, veterinary medicinal products or solvents are used during the entire production process.

The applicant reported that the feed used to feed L. migratoria is a plant-derived material compliant with Directive 2002/32/EC2 and produced according to Good Manufacturing Practice (GMP+).

During farming, L. migratoria can be infected by pathogens, including bacteria, viruses, entomopathogenic fungi, microsporidia and protists (Eilenberg et al., 2015). Pathogens that may affect L. migratoria include the virus Cricket iridovirus (CrIV) (Kleespies et al., 1999), the fungus Metarhizium acridum (Eilenberg et al., 2015) and a microsporidium named Paranosema locustae (Maniania et al., 2008). Literature review conducted by the applicant highlighted that these pathogens are specific at species or family level, and non-pathogenic for humans or other vertebrates. An example of food-borne bacteria to which L. migratoria is sensitive is Bacillus thuringiensis (Song et al., 2008). However, its potential presence in the NF is monitored by microbiological analysis of Bacillus as reported in Section 7 Table 4. In fact, B. cereus and B. thuringiensis are considered indistinguishable in the context of clinical diagnostics and food microbiology (EFSA BIOHAZ Panel, 2016).

Adults are harvested (3–5 weeks old) after being separated from the substrate and faeces. Decayed insects are identified by visual inspection and removed from the rearing batches. After the harvest, a minimum of 24 h fasting step is implemented to allow the adults to discard their bowel contents.

The post-harvesting processing includes killing of the adults by freezing and storing at –18°C. Three formulations of the NF are then obtained by processing. The formulation ‘LM frozen’ is obtained after removing of legs and wings, rinsing in water, blanching in hot water (> 90°C for at least 10 min) and freezing. The formulation ‘LM dried’ is obtained after removing legs and wings, rinsing in water, blanching in hot water (> 90°C for at least 10 min) and freeze-drying. The formulation ‘LM powder’ is obtained after rinsing in water, blanching in hot water (> 90°C for at least 10 min), freeze-drying and grinding (including legs and wings).

The thermal treatment contributes to the reduction of the microbial load of the NF as well as the elimination of potentially present viruses, parasites and reduction of enzymatic activity. Dehydration of the insects takes place in freeze dryers, resulting in a final product with moisture < 5%. Body parts (i.e. legs and wings) are removed by the manufacturer (LM frozen and LM dried) to reduce the risk of intestinal constipation that could be possibly caused by ingestion of the large spines on the insect tibia (FAO, 2013a). The NF LM powder is obtained via mechanical grinding of the insect (with legs and wings) and sieving to reduce particle size below 1 mm. The formulations of the NF are packed in hermetically closed packaging and stored at –18°C (LM frozen), or at room temperature (LM dried, LM powder).

The Panel considers that the production process is sufficiently described.

3.4 Compositional data

In order to confirm that the manufacturing process is consistent and adequate to produce on a commercial scale a product with certain characteristics, the applicant provided qualitative and quantitative data on chemical and microbiological parameters for a number of different batches of the NF formulations i.e. a) LM frozen; b) LM dried; c) LM powder. For all parameters, at least five independently produced batches were analysed. Considering the production process, the Panel accepted the view that the two formulations of the NF (LM frozen and LM dried) are representative of each other regarding the compositional parameters, when the difference in moisture is taken into account. Where indicated, analyses were performed on LM dried including legs and wings and considered as representative of LM powder. Microbiological analyses were performed on all formulations of the NF.

Certificates of accreditation of the laboratories that conducted the analyses were provided by the applicant. Analytical data were produced using methods validated for other types of matrices. Whenever in-house methods were employed, a full description of the method as well as results of the respective validation procedures have been provided.

The NF is a ‘whole food’ as defined by EFSA Scientific Committee (2011), meaning that all its constituents cannot be fully identified and/or characterised (EFSA NDA Panel, 2016).

The results of the proximate analysis of the NF are presented in Table 1. The amino acid, fatty acid, vitamin and mineral composition are reported in Section 15.

The Panel notes that there is a variation of the values of some proximate parameters, but this can be expected due to analytical variation and since the NF is produced using whole insects. The compositional values may also depend on the rearing conditions (feed, developmental stage at the time of harvesting, ambient conditions (Oonincx and van der Poel, 2011; Rumpold and Schlüter, 2013a).

Regarding the crude protein content of the NF, the Panel notes that Janssen et al. (2017) suggest that it is possibly overestimated when using the nitrogen-to-protein conversion factor of 6.25, mainly due to the presence of chitin. This issue will be addressed in detail in Section 15.

Chitin is the main form of dietary fibre in L. migratoria (Oonincx and van der Poel, 2011; Hahn et al., 2018). It is a linear polysaccharide constituted by β-(1,4)-linked 2-amino-2-deoxy-β-D-glucopyranose and 2-acetamido-2-deoxy-β-D-glucopyranose residues (Roberts, 1992). Due to the differences in body parts, after EFSA's request, the applicant provided analytical data on the levels of chitin in 5 independently produced batches in two formulations of the NF (LM dried and LM powder). The panel notes that a nationally or internationally recognised reference method for the analytical determination of chitin does not exist. The chitin content in the NF was based on the protocol described by Hahn et al. (2018), in which chemical treatment based on Acid Detergent Fibre – Acid Detergent Lignin is used to estimate the chitin content in different insects. The Panel considers that the differences between the content of dietary fibre (Table 1) and chitin (Table 2) are due to different analytical methods utilised.

Levels of heavy metals in LM powder are reported in Table 3. The applicant compared the values to the maximum levels for other foods as set in Regulation (EC) No. 1881/2006. The Panel notes that the levels of heavy metals reported for the NF are comparable to those set for other foods, and that in the current EU legislation, no maximum levels of heavy metals are set for insects as food.

Analytical data on the concentrations of aflatoxins B1, B2, G1, G2, ochratoxin A, nivalenol, deoxynivalenol, zearalenone, T2- and HT2-toxins and, upon EFSA's request, fumonisin B1 and fumonisin B2, were provided (Table 3). Values were compared to maximum limits for different foods set in EC Regulation (EC) No 1881/2006. The Panel notes that, in the current EU legislation, no maximum levels of mycotoxins are set for insects as food.

The content of dioxins and dioxins-like compounds were provided by the applicant (Table 3) and values were compared to maximum levels for other foods as set in Regulation (EC) No 1881/2006). The Panel notes that in the current EU legislation, no maximum levels of dioxins-like compounds are set for insects as food.

Analytical data of the pesticide concentrations for five independently produced batches of the NF have been provided. The results showed that the tested pesticide concentrations in the LM powder are below the limits of quantification (LOQ) of the implemented method used (GC-MS ITD Equal CEN/TR 16468 and LC-MS Equal CEN/TR 15641) and are complying with Regulation (EC) No 396/20053 defining maximum residue limits (MRL) of pesticides in foods.

Given the vegetable origin of the feeding substrate and the absence of prion or prion-related encoding genes in insects, the development of specific prion diseases due to the consumption of the NF is not expected (EFSA Scientific Committee, 2015).

The applicant provided analytical data for histamine for five independently produced batches of LM dried and LM powder (all below 10 mg/kg) and values were compared to the limit of 200 mg/kg for histamine in fishery products set in Commission Regulation EC No 2073/2005. High concentrations of putrescine (470–620 mg/kg in LM dried and 279–299 mg/kg in LM powder) were reported. No legal limit has been established for putrescine in any food although it may accumulate at very high concentration in cheese (up to 1,560 mg/kg), fermented sausages (up to 1,550 mg/kg) and fish sauces (up to 1,220 mg/kg) (EFSA BIOHAZ Panel, 2011). A recent study by del Rio et al. (2018) described a real-time analysis of the cytotoxicity of putrescine and cadaverine on intestinal cell cultures and found that the LOAEL (lowest observed adverse effect level) for putrescine was 881.50 mg/kg and for cadaverine 510.89 mg/kg. Formation of biogenic amines can occur by endogenous biosynthesis, uptake from the feed source and by bacteria of the intestinal microbiota of insects. It can also occur during food processing and storage as result of bacterial contamination (EFSA BIOHAZ Panel, 2011). Upon EFSA's request, the applicant was asked to analyse the NF for Pseudomonas aeruginosa, which could also be responsible for biogenic amines production. All formulations of the NF were tested and levels of < 10 cfu/g were reported.

The applicant provided microbiological data on five independently produced batches of all NF formulations (LM frozen, LM dried, LM powder) (Table 4).

The Panel notes that the applicant did not provide the actual values of the microbiological parameters, but instead the quantification limits as defined by the dilutions used upon the analyses. Furthermore, the Panel notes that the microbiological values of the analysed samples do not exceed the given specification limits.

The Panel considers that information provided on the composition is sufficient for characterising the NF.

3.5 Specifications

The specifications of the NF as proposed by the applicant are indicated in Table 7.

The Panel considers that the information provided on the specifications of the NF is sufficient and does not raise safety concerns.

3.6 History of use of the NF and/or of its source

L. migratoria either collected from the wild or reared in farms is consumed as part of the customary diet in several non-EU countries worldwide. Human consumption of L. migratoria has largely been documented in Madagascar, Cameroun, Congo, Zimbabwe, Sudan, South Sudan, Papua New Guinea, Thailand, China and Morocco (Jongema, 2017). Species of locusts are considered the most consumed insect species in Central African Republic (Durst et al., 2010). L. migratoria is commonly consumed as snack, side dish and in cooking sauces. Their preparation includes frying, roasting, boiling and sun drying and legs and wings are removed before consumption.

Since 2012, several companies and specialised shops have been selling L. migratoria in the EU either as whole food or by adding it to other food products. Since the 1 May 2017, L. migratoria is among the insect species that can be legally introduced in the Swiss market as food, when commercially reared.

L. migratoria can be found in the Dutch market since 2016.

3.7 Proposed uses and use levels and anticipated intake

3.8 Absorption, distribution, metabolism and excretion (ADME)

Not relevant.

3.9 Nutritional information

The applicant provided nutritional analyses of the NF formulations which consist mainly of protein, fat, dietary fibre (mainly chitin) and inorganic matter (LM dried, LM powder); water, protein, fat, dietary fibre (mainly chitin) and inorganic matter (LM frozen). The energy value of the NF is on average 690, 2,250 and 2,350 kJ/100 g, respectively, for LM frozen, LM dried and LM powder (Table 1). Analytical data on the amino acid composition, the fatty acid content, minerals and vitamins in the NF formulations have been provided for five batches of LM powder (LM dried with legs and wings).

The NF contains on average 14.4 g crude protein per 100 g LM frozen, 48.6 g crude protein per 100 g LM dried and 55.0 g crude protein per 100 g LM powder, calculated using a protein conversion factor of 6.25. The Panel notes that the use of the conventional factor overestimates the levels of true protein content in L. migratoria due to the presence of considerable amounts of non-protein nitrogen derived mainly from chitin (Boulos et al., 2020). A study provided by the applicant identified a nitrogen-to-protein conversion factor of 5.31 for LM powder (LM dried with legs and wings). Using this factor, the protein content of the NF is about 15% lower than considering a conversion factor of 6.25. For regulatory purposes for nutritional labelling, protein is defined as the total nitrogen measured by the Kjeldahl method multiplied by a nitrogen-to-protein conversion factor of 6.25 [Regulation (EU) No 1169/2011 on the provision of food information to consumers].

The applicant quantified the amino acids in five batches of the NF according to ISO 13903:2005 and EU 152/2009 (Appendix 34). The applicant also analysed the amount of amino acids in g per 100 g protein of LM powder (LM dried with legs and wings). Results show that in the protein from LM powder, all essential amino acids including sulfur containing amino acids were present in quantities similar or higher than the recommended levels by FAO (2013b) (Appendix 34). Furthermore, the applicant conducted a study of the true ileal protein digestibility during transit through the dynamic in vitro gastrointestinal model (tiny-TIM). Casein was used as a reference protein. The tests were conducted by an accredited laboratory in accordance with GLP. The nitrogen digestibility was expressed as a percentage of the total nitrogen intake, including non-protein nitrogen. As result, the true ileal nitrogen digestibility of LM powder (LM dried with legs and wings) was 55.4% ± 2.4%, compared to casein 75.3% ± 1.4%, indicating that proteins of L. migratoria are less bio-accessible than casein. Following the recommendation by FAO, 2013b, the protein quality was determined by the Digestible Indispensable Amino Acid Score (DIAAS), using the reference value of child, adolescent, adult group (3–10 years). The DIAAS value for LM powder corresponded to 70%, compared to casein with a DIAAS of 91%. Sulfur amino acids (methionine + cysteine) were the limiting amino acids.

The major fatty acids in LM powder (LM dried with legs and wings) are palmitic acid, stearic acid, oleic acid, linoleic acid and alpha linolenic acid (Appendix 36). On average, saturated, monounsaturated fats and polyunsaturated fatty acids (linoleic and alpha-linolenic acid) constitute 38.2%, 39.9% and 21.9% of total fatty acids, respectively. Similar fatty acids profile for L. migratoria is reported in literature (Clarkson et al., 2018). The average trans fatty acid content is 0.4% fat.

The applicant provided analytical data on the levels of some minerals and vitamins (Table 10), and after EFSA's request, boron, molybdenum, iodine and selenium. LM powder (LM dried with legs and wings) contains riboflavin, niacin, pantothenic acid, biotin and cobalamin besides vitamin E and small amounts of thiamine, folic acid, retinol and vitamin D. It also contains P, Zn, Cu and Mn.

Considering the mean contents reported in Table 10, values reported in the specifications, and the estimated P95 of exposure to the NF, the Panel notes that none of the existing upper levels for the analysed micronutrients are expected to be exceeded, for any population groups.

It has been reported that chitin can be partially digested in the human stomach by the acidic mammalian chitinase (AMCase) (Paoletti et al., 2009; Muzzarelli et al., 2012). However, Paoletti et al. (2009) suggested that reduction of chitin intake in western diets may have led to reduced expression of chitinase genes, thus resulting in loss of catalytic efficacy. The NF contains on average 1.8 g, 6.5 g and 11.7 g chitin in 100 g LM in frozen, dried and powder formulations, respectively (see Table 2). The Panel considers that chitin is an insoluble fibre that is not expected to be digested in the small intestine of humans to any significant degree. It is also rather resistant to microbial fermentation and therefore assumed to be excreted mainly unchanged. Additionally, the Panel notes that chitin can bind bivalent minerals (Franco et al., 2004; Anastopoulos et al., 2017) possibly affecting their bioavailability, as reported for dietary fibres in general (Baye et al., 2017).

Insects may contain antinutritional factors (ANFs) such as tannins, oxalates, phytate, hydrogen cyanide (Jonathan et al., 2012; Shantibala et al., 2014), thiaminases (Nishimune et al., 2000) and protease inhibitors (Eguchi, 1993). The applicant determined the concentrations of total polyphenols, tannins, oxalic acid, phytic acid, hydrogen cyanide and trypsin inhibitors in five independently produced batches of LM powder (LM dried with legs and wings) (Table 11). The reported values in the NF are comparable to the occurrence levels of these compounds in other foodstuffs (Rao and Prabhavathi, 1982; Gupta, 1987; Holmes and Kennedy, 2000; Schlemmer et al., 2009; EFSA CONTAM Panel, 2019).

The Panel considers that taking into account the composition of the NF and the proposed conditions of use, the NF is not nutritionally disadvantageous.

3.10 Toxicological information

Some insect species secrete chemical substances with potentially toxic effects, as part of their defence mechanisms (Dzerefos et al., 2013; Rumpold and Schlüter, 2013b). However, regarding L. migratoria the production of such substances has not been reported in the literature.

Regarding the safety of chitin present in the NF, the applicant referred to EFSA's scientific opinion on the safety of ‘chitin-glucan’ as a Novel Food ingredient (EFSA NDA Panel, 2010). However, the Panel considers that the polymer chitin-glucan cannot be considered as representative of the chitin derived from L. migratoria.

Potential adverse health effects of chitin may be related to immunological effects. As reviewed by Komi et al. (2018), chitin has been shown to activate a variety of innate (eosinophils, macrophages) and adaptive immune cells (IL-4/IL-13 expressing T helper type-2 lymphocytes) and this implies the potential to promote immunological reactions including hypersensitivity. EFSA identified an article (Niho et al., 1999) (Japanese language, only abstract available in English) stating that no toxic effects related to chitin were observed in F344 rats at concentrations up to 5% of chitin in the diet for 13 weeks. No firm conclusion could be drawn by the Panel since only the abstract was accessible.

The applicant has taken commitment for two toxicity studies to be performed with the aqueous extract from NF. However, due to the insolubility of the test item only one study assessing cytotoxicity was performed.

In addition, the applicant provided a study retrieved from literature (Turkez et al., 2014) which investigates the in vitro genotoxic potential of water soluble extracts of L. migratoria on cultured human blood cells. Additional publications were also identified by EFSA describing subacute and subchronic toxicity studies performed in the rat with powdered locust mixed in the diet (up to 13 weeks; Ochiai et al., 2020) or administered by gavage (28 days; Kwak et al., 2020).

3.11 Allergenicity

A literature search has been carried out by the Applicant using Google Scholar and Scopus^® to retrieve relevant data. The relevant information on the source, production process, protein characterisation, reported case studies for allergenicity due to exposure or consumption of L. migratoria, immunological studies, cross-reactivity and effect of processing/digestion on allergens have been reported.

Locusta migratoria (family Acrididae) belongs to the subphylum Hexapoda (class Insecta), one of the four subphyla of Arthropoda, the others being Crustacea, Myriapoda and Chelicerata. Within arthropods, several allergens have been reported, including tropomyosin (Reese et al., 1999), arginine kinase (Binder et al., 2001) and glutathione S-transferase (Galindo et al., 2001). Furthermore, chitinases, the enzymes that degrade chitin, have been identified as an allergen in some insect species (Zhao et al., 2015).

Few prevalence studies on food allergy related to insects, mainly for Asian populations, are available (China and Laos) (Ji et al., 2009; Barennes et al., 2015). Sokol et al. (2017) registered two events of anaphylaxis due to ingestion of chapulines (roasted grasshopper from Oaxaca, Mexico) in patients allergic to crustaceans who had no previous exposure to grasshoppers. According to Ji et al. (2009), ingestion of insects was the cause of 18% anaphylactic food-related reactions in China from 1980 to 2007, and locusts accounted for 27 cases out of 358. Furthermore, the occurrence of anaphylactic reactions due to fried insects (grasshoppers and crickets) has also been registered in Thailand (Piromrat et al., 2008).

Multiple allergens have been found in L. migratoria extracts of different molecular weight (Tee et al., 1988; Lopata et al., 2005; Ji et al., 2009) using different approaches. By using a proteomic and bioinformatic approach, Barre et al. (2021) identified 73 proteins in L. migratoria, corresponding to pan-allergens which develop cross-reactivity with other homologous proteins present in arthropods (house dust mites and crustaceans), followed by allergens from molluscs and nematodes. These include arginine kinase, chitinase, glutathione S-transferase, HSP 70, hexamerin, serine protease, tropomyosin and trypsin. Tropomyosin appeared as a major pan-allergen largely distributed among dust mites, insects, crustaceans and molluscs, as also confirmed by Sokol et al. (2017).

Cross-reactivity to L. migratoria protein extracts has been evidenced by immunoblotting in sera from crustaceans and house dust mite allergic individuals (Pali-Schöll et al., 2019a) and in sera from shrimp allergic individuals (Broekman et al., 2017). The main reason for cross-reactivity is the high protein homology between phylogenetically related organisms, being evident not only between species within the same subphylum, but also between species from different arthropod subphyla. It includes crustacean species (e.g. shrimp, crab), chelicerates (e.g. mites) and several insect species (Santos et al., 1999; Binder et al., 2001; Galindo et al., 2001; Liu et al., 2009; Lopata et al., 2010; Verhoeckx et al., 2014; Van Broekhoven et al., 2016; Rougé and Barre, 2017; Broekman et al., 2017; De Gier and Verhoeckx, 2018).

In addition, food processing per se may also have an influence on allergenicity, and this applies to insect allergens as well, although the effect of food processing on allergenicity cannot be predicted (EFSA NDA Panel, 2014). Pali-Schöll et al. (2019b) reported that some processing treatments, such as enzymatic hydrolysis or thermal treatments reduced the IgE binding from crustacean and mite allergic patients to L. migratoria (immunoblotting and skin prick test). de Gier and Verhoeckx (2018) reported a review of the literature on the effect of thermal processing on allergenicity of several insect proteins and concluded that it did not eliminate insect protein allergenicity.

Additional aspects should be taken into consideration depending on the feed substrate used to rear L. migratoria, as it might include common allergenic foods. The applicant reported that a plant-based substrate containing gluten was used as feed, hence traces of gluten may be found in the insects’ gut. The Panel notes that changes in the feed can possibly introduce additional allergens, including allergens which require mandatory labelling according to Annex II of Regulation (EU) No 1169/2011, since traces of the allergens may remain in the gut of L. migratoria despite implementing a fasting step.

A frequently reported case of allergic symptoms to insects, including L. migratoria, relates to occupational exposure (Burge et al., 1980; Tee et al., 1988; Lopata et al., 2005).

The Panel considers that the consumption of the NF might trigger primary sensitisation to L. migratoria proteins. The Panel also considers that allergic reactions may occur in subjects allergic to crustaceans, mites and molluscs (cross-reactivity). Furthermore, Panel notes that additional allergens may end up in the NF, if these allergens are present in the substrate fed to the insects.

5.1 Protection of Proprietary data in accordance with Article 26 of Regulation (EU) 2015/2283

The Panel could not have reached the conclusion on the safety of the NF under the proposed conditions of use without the data claimed as proprietary by the applicant (description of the production process, analytical data on the composition of the NF, analytical data on contaminants in the NF, stability and microbiological status, data on NF sales, intake assessment, protein digestibility and DIAAS, genotoxicity and cytotoxicity study).

Information provided in this Annex is shown in an Excel file (downloadable at https://efsa.onlinelibrary.wiley.com/doi/10.2903/j.efsa.2021.6667#support-information-section).