Development and evaluation of real-time PCR targets for the authentification of insect flours

• Debode, F. , Marien, A. , Akhatova, D. , Khol, C. , Sedefoglu, H. , Mariscal-Diaz, L. , Gérard, A. , Fumière, O. , Maljean, J. , Hulin, J. , Francis, F. & Berben, G. (2019). Development and evaluation of real-time PCR targets for the authentification of insect flours. Wageningen Academic Publishers. Proceedings in: 70th EAAP Annual Meeting, Ghent (Belgium), 26-30 August, 325.

Type	Conference Proceedings
Year of conference	2019
Title	Development and evaluation of real-time PCR targets for the authentification of insect flours
Editor	Wageningen Academic Publishers
Conference name	70th EAAP Annual Meeting
Conference location	Ghent (Belgium)
Volume	25
Pages	325
conference Date	26-30 August
ISSN	ISSN 1382-6077
Endnote keywords	insects, real time PCR
Abstract	Insects are rich in proteins and could be an alternative source of proteins to feed animals. In recent years, numerous companies have started producing insects for feed purposes. In Europe, the processed animal proteins obtained from seven insect species namely Black Soldier Fly (Hermetia illucens), Common Housefly (Musca domestica), Yellow Mealworm (Tenebrio molitor), Lesser Mealworm (Alphitobius diaperinus), House Cricket (Acheta domesticus), Banded Cricket (Gryllodes sigillatus) and Field Cricket (Gryllus assimilis) have been authorised for aquaculture by Commission Regulation (EU) 2017/893 since 1 July 2017. The stakeholders are now hoping future modifications of the legislation to allow the feeding of poultry and pig. For further authorization in Europe, many questions must be clarified concerning the presence of antinutritional compounds, the risk associated to pathogens, to residues (pesticides, antibiotics, heavy metals) and to allergens. Methods to detect if a product really contains insects and to authenticate insect products will also be mandatory. European Commission Regulation No 51/2013 named the Polymerase Chain Reaction as a reference method to determine the constituents of animal origin in feed. To fulfill the need in authentication methods, we started to develop real-time PCR methods specific to the insects listed above. Methods for Tenebrio molitor (Debode et al., 2017) and Hermetia illucens (Marien et al., 2018) were already developed and published. Methods are now ready for Alphitobius diaperinus, Gryllus assimilis, Acheta domesticus and Musca domestica. These methods were checked with success for their specificity against more than 40 insect species but also against a large number of mammals, birds, fishes, crustaceans and plants. The methods reached the recommended performance criteria for sensitivity (limit of detection ? 20 copies of the target) and robustness (slight modifications brought to the standard protocol). The applicabilily of the methods was also tested on real industrial insect flours at the exception of Musca domestica as industrial samples were not available. An interrogation also remains for Musca domestica as many close species are erroneously considered to be Musca domestica. In addition to real-time PCR, approaches based on metagenomics and high throughput sequencing are ongoing in order to provide a more complete overview of the insects in presence.
Fichier
Authors	Debode, F., Marien, A., Akhatova, D., Khol, C., Sedefoglu, H., Mariscal-Diaz, L., Gérard, A., Fumière, O., Maljean, J., Hulin, J., Francis, F., Berben, G.