Development of real-time PCR tests for the detection of Tenebrio molitor in food and feed

Insect; Tenebrio molitor; Animals; Error detection; Genes; Proteins; Limit of detection; Performance criterion; Processed animal proteins; Qualitative method; Real-time PCR; Real-time PCR method; Polymerase chain reaction; DNA; DNA 18S; RNA 18S; RNA 28S; Article; Bombyx mori; Gammarus; Zophobas morio; Tenebrio; Animal Feed; Food Analysis; Real-Time Polymerase Chain Reaction

Abstract :

[en] Insects are rich in proteins and could be an alternative source of proteins to feed animals and humans. Numerous companies have started the production of insects for feed purposes. In Europe, these processed animal proteins are not yet authorised by legislation as many questions still need to be answered concerning this ‘novel food’. Authorisations will be possible when methods of authentication of the products are available. In this study we propose real-time PCR methods for the specific detection of the mealworm (Tenebriomolitor), one of the most widely used insects for food and feed production. Two PCR assays are proposed: the first based on the wingless gene and the second based on the cadherin gene. The PCR tests amplify fragments of 87 bp. These qualitative methods were tested according to several performance criteria. The specificity was tested on 34 insect species’ DNA, but also on non-insect species including crustacean, mammals, birds and plants. The limit of detection was determined and was below 20 copies for the two PCR tests. The applicability of the tests was demonstrated by the analysis of real-life processed samples containing T. molitor. © 2017 Informa UK Limited, trading as Taylor & Francis Group.

Disciplines :

Food science

Author, co-author :

Debode, F.; Unit Traceability and Authentication, Walloon Agricultural Research Center (CRA-W), Gembloux, Belgium

Marien, A.; Unit Traceability and Authentication, Walloon Agricultural Research Center (CRA-W), Gembloux, Belgium, European Union Reference Laboratory for Animal Proteins in feedingstuffs (EURL-AP), Gembloux, Belgium

Gerard, Amaury ; Université de Liège - ULiège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Chimie des agro-biosystèmes

Francis, Frédéric ; Université de Liège - ULiège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Gestion durable des bio-agresseurs

Fumière, O.; Unit Traceability and Authentication, Walloon Agricultural Research Center (CRA-W), Gembloux, Belgium, European Union Reference Laboratory for Animal Proteins in feedingstuffs (EURL-AP), Gembloux, Belgium

Berben, G.; Unit Traceability and Authentication, Walloon Agricultural Research Center (CRA-W), Gembloux, Belgium, European Union Reference Laboratory for Animal Proteins in feedingstuffs (EURL-AP), Gembloux, Belgium

Language :

English

Title :

Development of real-time PCR tests for the detection of Tenebrio molitor in food and feed

Publication date :

2017

Journal title :

Food Additives and Contaminants. Part A. Chemistry, Analysis, Control, Exposure and Risk Assessment

ISSN :

1944-0049

eISSN :

1944-0057

Publisher :

Taylor & Francis, United States

Volume :

Issue :

Pages :

1421-1426

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 19 August 2018

Statistics

Number of views

156 (10 by ULiège)

Number of downloads

517 (6 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

AFNOR XP-V-03-020-2. 2008. Détection et quantification des organismes végétaux génétiquement modifiés et produits dérivés. Partie 2:Méthodes basées sur la réaction de polymérisation en chaîne. Saint-Denis La Plaine, France:Association Française de Normalisation.
Brower AVZ, DeSalle R., 1998. Patterns of mitochondrial versus nuclear DNA sequence divergence among nymphalid butterflies:the utility of wingless as a source of characters for phylogenetic inference. Insect Mol Biol. 7:73–82.
Cameron SL. 2014. Insect mitochondrial genomics:implications for evolution and phylogeny. Annu Rev Entomol. 59:95–117.
Carapelli A, Frati F, Nardi F, Dallai R, Simon C. 2000. Molecular phylogeny of the apterygotan insects based on nuclear and mitochondrial genes. Pedobiologia. 44:361–373.
Cook LG, Gullan PJ, Trueman HE. 2002. A preliminary phylogeny of the scale insects (Hemiptera:sternorrhyncha:coccoidea) based on nuclear small-subunit ribosomal DNA. Mol Phylogenet Evol. 25:43–52.
Dawnay N, Ogden R, McEwing R, Carvalho GR, Thorpe RS. 2007. Validation of the barcoding gene COI for use in forensic genetic species identification. Forensic Sci Int. 173:1–6.
Deagle BE, Jarman SN, Coissac E, Pompanon F, Taberlet P. 2014. DNA metabarcoding and the cytochrome c oxidase subunit I marker:not a perfect match. Biol Lett. 10:20140562.
Debode F, Janssen E, Berben G. 2007. Physical degradation of genomic DNA of soybean flours does not impair relative quantification of its transgenic content. Eur Food Res Technol. 226:273–280.
Debode F, Janssen E, Marien A, Berben G. 2012. DNA detection by conventional and real-time PCR after extraction from vegetable oils. J Am Oil Chemists’ Soc. 89:1249–1257.
Debode F, Marien A, Janssen E, Bragard C, Berben G. 2017. The influence of amplicon length on real-time PCR results. Biotechnologie, Agronomie, Société et Environnement. 21:3–11.
EURL-AP Standard Operating Procedure. 2013. Detection of horse DNA using real-time PCR. Available from:http://eurl.craw.eu/img/page/sops/Protocol%20for%20detection%20of%20horse%20DNA%20using%20real-time%20PCR.pdf
EURL-AP Standard Operating Procedure. 2014. Detection of ruminant DNA in feed using real-time PCR. Available from:http://eurl.craw.eu/img/page/sops/EURL-AP%20SOP%20Ruminant%20PCR%20V1.1.pdf
Finke MD. 2002. Complete nutrient composition of commercially raised invertebrates used as food for insectivores. Zoo Biol. 21:269–285.
Ghaly AE, Alkoaik FN. 2009. The yellow mealworm as a novel source of protein. Am J Agric Biol Sci. 4:319–331.
Han SR, Lee BS, Jung KJ, Yu HJ, Yun EY, Hwang JS, Moon KS. 2016. Safety assessment of freeze-dried powdered Tenebrio molitor larvae (yellow mealworm) as novel food source:evaluation of 90-day toxicity in Sprague-Dawley rats. Regul Toxicol Pharmacol. 77:206–212.
Hasegawa E, Kasuya E. 2006. Phylogenetic analysis of the insect order Odonata using 28S and 16S rDNA sequences:a comparison between data sets with different evolutionary rates. Entomol Sci. 9:55–66.
Häusling M, 2011. Report:the EU protein deficit:what solution for a long-standing problem? European Parliament. Committee on Agriculture and Rural Development. A7-0026/2011, Brussels, Belgium.
Hebert PD, Cywinska A, Ball SL. 2003b. Biological identifications through DNA barcodes. Proc Royal Soc London B:Biol Sci. 270:313–321.
Hebert PD, Ratnasingham S, de Waard JR. 2003a. Barcoding animal life:cytochrome c oxidase subunit 1 divergences among closely related species. Proc Royal Soc London B:Biol Sci. 270:S96–S99.
Hillis DM, Dixon MT. 1991. Ribosomal DNA:molecular evolution and phylogenetic inference. Q Rev Biol. 66:411–453.
Hoogendoorn M, Heimpel GE. 2001. PCR‐based gut content analysis of insect predators:using ribosomal ITS‐1 fragments from prey to estimate predation frequency. Mol Ecol. 10:2059–2067.
ISO 21571. 2005. Foodstuffs–Methods of analysis for the detection of genetically modified organisms and derived products–Nucleic acid extraction. Geneva:International Organisation for Standardization.
Jinbo U, Kato T, Ito M. 2011. Current progress in DNA barcoding and future implications for entomology. Entomol Sci. 14:107–124.
Jones LD, Cooper RW, Harding RS. 1972. Composition of mealworm Tenebrio molitor larvae. J Zoo Anim Med. 3:34–41.
Kim MI, Wan X, Kim MJ, Jeong HC, Ahn NH, Kim KG, Han YS, Kim I. 2010. Phylogenetic relationships of true butterflies (Lepidoptera:papilionoidea) inferred from COI, 16S rRNA and EF-1α sequences. Mol Cells. 30:409–425.
Kjer KM. 2004. Aligned 18S and insect phylogeny. Syst Biol. 53:506–514.
Kutyavin IV, Afonina IA, Mills A, Gorn VV, Lukhtanov EA, Belousov ES, Singer MJ, Walburger DK, Lokhov SG, Gall AA, et al. 2000. 3′-minor groove binder-DNA probes increase sequence specificity at PCR extension temperatures. Nucleic Acids Res. 28:655–661.
Li X, Li J, Zhang S, He Y, Pan L. 2013. Novel real-time PCR method based on growth hormone gene for identification of Salmonidae ingredient in food. J Agric Food Chem. 61:5170–5177.
Malewski T, Draber-Mońko A, Pomorski J, Łoś M, Bogdanowicz W. 2010. Identification of forensically important blowfly species (Diptera:calliphoridae) by high-resolution melting PCR analysis. Int J Legal Med. 124:277–285.
Mandal SD, Chhakchhuak L, Gurusubramanian G, Kumar NS. 2014. Mitochondrial markers for identification and phylogenetic studies in insects–A Review. DNA Barcodes. 2:1–9.
Ng WK, Liew FL, Ang LP, Wong KW. 2001. Potential of mealworm (Tenebrio molitor) as an alternative protein source in practical diets for African catfish, Clarias gariepinus. Aquaculture Res. 32:273–280.
Pons J. 2006. DNA‐based identification of preys from non‐destructive, total DNA extractions of predators using arthropod universal primers. Mol Ecol Notes. 6:623–626.
Pons J, Barraclough TG, Gomez-Zurita J, Cardoso A, Duran DP, Hazell S, Kamoun S, Sumlin WD, Vogler AP. 2006. Sequence-based species delimitation for the DNA taxonomy of undescribed insects. Syst Biol. 55:595–609.
Porter TM, Gibson JF, Shokralla S, Baird DJ, Golding GB, Hajibabaei M. 2014. Rapid and accurate taxonomic classification of insect (class Insecta) cytochrome c oxidase subunit 1 (COI) DNA barcode sequences using a naïve Bayesian classifier. Mol Ecol Resour. 14:929–942.
Ramos-Elorduy J, González EA, Hernández AR, Pino JM. 2002. Use of Tenebrio molitor (Coleoptera:tenebrionidae) to recycle organic wastes and as feed for broiler chickens. J Econ Entomol. 95:214–220.
Roger AJ, Sandblom O, Doolittle WF, Philippe H. 1999. An evaluation of elongation factor 1 alpha as a phylogenetic marker for eukaryotes. Mol Biol Evol. 16:218–233.
Rumpold BA, Schlüter OK. 2013. Nutritional composition and safety aspects of edible insects. Mol Nutr Food Res. 57:802–823.
Sheppard SK, Bell J, K D S, Fenlon J, Skervin D, Symondson WOC. 2005. Detection of secondary predation by PCR analyses of the gut contents of invertebrate generalist predators. Mol Ecol. 14:4461–4468.
Siemianowska E, Kosewska A, Aljewicz M, Skibniewska KA, Polak-Juszczak L, Jarocki A, Jedras M. 2013. Larvae of mealworm (Tenebrio molitor L.) as European novel food. Agric Sci. 4:287.
Simon S, Schierwater B, Hadrys H. 2010. On the value of Elongation factor-1α for reconstructing pterygote insect phylogeny. Mol Phylogenet Evol. 54:651–656.
Virgilio M, Backeljau T, Nevado B, De Meyer M. 2010. Comparative performances of DNA barcoding across insect orders. BMC Bioinformatics. 11:1.
Wells JD, Škaro V. 2014. Application of DNA-Based Methods in Forensic Entomology. Forensic DNA Applications:an Interdisciplinary Perspective 353. ISBN:978-1-4665-8022-0. New York:CRC Press, Taylor and Francis group.
Yu DW, Ji Y, Emerson BC, Wang X, Ye C, Yang C, Ding Z. 2012. Biodiversity soup:metabarcoding of arthropods for rapid biodiversity assessment and biomonitoring. Methods Ecol Evol. 3:613–623.