Edible insects are rich in protein and can serve as a significant source of vitamins and minerals. The house cricket ( Acheta domesticus ) stands out as one of the most nutritious edible insects. In various parts of the world, crickets are consumed roasted, baked, fried, boiled and in the form of cricket flour, a powder of dried and ground crickets. In Europe, processed animal proteins derived from eight insect species (including A. domesticus ) have received authorization for use in fish, pig and poultry feed. Therefore, the development of a method for detecting house crickets is essential to verify product compliance and provide accurate labelling information to the end user. In this study, we have established a real-time PCR assay for the specific detection of house crickets. This method is based on the amplification of a mitochondrial fragment that codes for cytochrome b. We assessed its specificity by testing it against 39 other insect species, 7 plant species and 31 other animal species. Furthermore, we successfully evaluated the amplification efficiency, sensitivity, robustness, applicability on commercial samples and transferability to a second laboratory.

The challenges for the feed analysis sector will be to ensure feed quality and safety. Innovative strategies need to be implemented to authenticate feed and feed ingredients and to check that they fulfill all labelling obligations. This chapter focusses on the authentication of feed by different but complementary analytical techniques as microscopy, vibrational spectroscopy, genomics and proteomics. For each technique, trends are presented and, in addition, some combinations of technologies are shown in order to find solutions for complex situations. Moreover, with the explosion of data provided by the different technologies, Data Science is gaining importance as well as the interest for fast, mobile and greener technologies.

For insect-based formulations, food analysis is essential to ensure authenticity. According to the spread of the real-time-polymerase chain reaction technique, it started to be tested for entomological occurrence in food with limited success according to the insect species to be detected. Mass spectrometry-based proteomic methods are increasingly being used for the specific detection of trace amounts in food products. Here, we focused on the potential use of proteomic analyses to detect four edible insect species namely Tenebrio molitor, Hermetia illucens, Alphitobius diaperinus and Gryllus assimilis in insect commercial flours. Reproducible results were obtained from liquid chromatography-mass spectrometry with heterogenous efficiency rates on insect identification according to the target species. Indeed, Tenebrio molitor and Gryllus assimilis were identified at species and genus level respectively whereas no efficient results at these taxonomic levels could be found for Hermetia illucens. Future applications of proteomics for edible insect detection in food are discussed in relation to available data and model.

Description du sujet. Cet article presente le developpement, lu0027evaluation et les limitations de tests de criblage bases sur des genes et utilises dans le cadre de la detection des OGM. Objectifs. Le but de la recherche est de proposer de nouvelles cibles basees sur la sequence codante de genes. Ces tests viendront rejoindre un panel du0027essais PCR deja existants de maniere a pouvoir realiser une meilleure detection des OGM. Methode. Des methodes basees sur la technique de PCR en temps reel avec sondes du0027hybridation ont ete developpees pour des genes frequemment utilises dans les constructions transgeniques. Les methodes ont ete evaluees sur la base de leur specificite et de leur sensibilite. Resultats. Huit tests PCR ont ete selectionnes sur la base des sequences des genes bar, pat, EPSPS, gox, gus et hsp70. Deux du0027entre eux ont montre un interet limite suite a lu0027obtention de signaux positifs lies a la presence du0027organismes donneurs. Ces interferences sont dues a des sequences residuelles du0027Escherichia coli dans les reactifs utilises pour lu0027amplification dans le cas du test gus, tandis que pour la cible hsp70, tenant compte quu0027actuellement le hsp70 est uniquement introduit dans des mais genetiquement modifies, le test est denue du0027interet car le hsp70 provient du mais. Les tests bar, pat, EPSPS et gox ont detecte avec succes les elements structuraux introduits dans les constructions transgeniques. Plusieurs tests ont ete proposes pour le gene EPSPS de maniere a couvrir les differentes versions du gene. Conclusions. Des tests PCR pour les sequences codantes des genes bar, pat, EPSPS et gox ont ete developpes et ont repondu aux criteres de performance souhaites en termes de specificite et de sensibilite. Les cibles peuvent etre amplifiees avec les memes conditions du0027amplification que celles du0027autres tests visant des promoteurs et des terminateurs, et peuvent donc etre utilisees en combinaison, de maniere a fournir une meilleure couverture des OGM ainsi que de premieres indications sur les OGM en presence.

Insects are rich in proteins and could be an alternative source of macronutrients to feed animals and humans. Over the past few years, numerous companies have started producing insects for feed purposes. In Europe, the processed animal proteins obtained from seven insect species have been authorised for aquaculture by Commission Regulation (EU) 2017/893 since 1 July 2017. Methods of authentication are required to check the conformity of the products. In this study, we propose a real-time PCR method for the specific detection of the black soldier fly (Hermetia illucens L.), one of the most widely used insects for feed production. The developed PCR assays amplify a 67 bp fragment based on the mitochondrial COX3 gene coding for subunit 3 of the cytochrome c oxidase. The qualitative method was tested according to several performance criteria. The specificity was tested against 51 insect species. The specificity was also checked against plant species and other animal species such as crustaceans, mammals and birds. The sensitivity, efficiency and robustness of the PCR test were successfully tested. The applicability of the test was proven through the analysis of real-life processed samples (industrial meals) of H. illucens.

Insects are rich in proteins and could be an alternative source of macronutrients to feed animals and humans. Over the past few years, numerous companies have started producing insects for feed purposes. In Europe, the processed animal proteins obtained from seven insect species have been authorised for aquaculture by Commission Regulation (EU) 2017/893 since 1 July 2017. Methods of authentication are required to check the conformity of the products. In this study, we propose a real-time PCR method for the specific detection of the black soldier fly (Hermetia illucens L.), one of the most widely used insects for feed production. The developed PCR assays amplify a 67 bp fragment based on the mitochondrial COX3 gene coding for subunit 3 of the cytochrome c oxidase. The qualitative method was tested according to several performance criteria. The specificity was tested against 51 insect species. The specificity was also checked against plant species and other animal species such as crustaceans, mammals and birds. The sensitivity, efficiency and robustness of the PCR test were successfully tested. The applicability of the test was proven through the analysis of real-life processed samples (industrial meals) of H. illucens.

This paper presents the inter-laboratory studies for the successful validation of qualitative real-time Polymerase Chain Reaction methods for the detection of Genetically Modified Organisms in food and feed. Two singleplex TaqMan methods were developed for the detection of the tE9 terminator present in several genetically modified events and the detection of the endogenous lectin gene from pea; the tE9 terminator donor organism. In addition, a TaqMan duplex method was validated, which allows the simultaneous detection of the pat and bar elements.The paper explains the different acceptance parameters used during the in-house validation, the method transferability study and finally the inter-laboratory validation, how this validation was conducted and the problems encountered. Statistical methods for the validation of the qualitative methods are also presented. This information can be used by laboratories for the verification of these methods during their implementation and for the validation of new methods.

Seen the growing number of genetically modified (GM) crops being developed, the need for cost- and time-effective detection methods is increasing to enable continuing the necessary effective control on food and feed products. This need can be achieved by performing an intensive screening combined with decision support tools like the CoSYPS matrix which permits reducing the number of events to be identified. To allow an extra covering power of the CoSYPS and to be able to include new EU-authorised GM events, two new SYBR®Green real-time PCR (qPCR) methods targeting two promoter sequences (pNOS and pFMV) were developed. These methods were validated using acceptance parameters such as the specificity, sensitivity and repeatability. In addition, the methods were transferred to a second laboratory, namely the Institute for Health and Consumer Protection, to test the reproducibility. Furthermore, the applicability and practicability of the methods were tested by using proficiency test samples. The two methods allow a specific and sensitive detection of the targets in food and feed samples and can be used efficiently in different laboratories.