Molecular approach for insect detection in feed and food: the case of Gryllodes sigillatus
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ORIGINAL PAPER
Molecular approach for insect detection in feed and food: the case of Gryllodes sigillatus Enrico Daniso1 · Francesca Tulli1 · Gloriana Cardinaletti1 · Roberto Cerri1 · Emilio Tibaldi1 Received: 1 April 2020 / Revised: 16 July 2020 / Accepted: 18 July 2020 © The Author(s) 2020
Abstract The production of insects on an industrial scale has attracted the attention of the research and agricultural industry as novel protein sources. To detect the presence of Gryllodes sigillatus (GS) in feed and food, a real-time PCR method based on the mitochondrial cytochrome b (CYB) gene is proposed by this study. Forty DNA samples of animal and plant origin were used to confirm the specificity of the qPCR system. The detection method’s performance was evaluated on different processed GS matrices including native GS (UnGS) and different commercial products: crunchy roasted samples (RoGS), insect meal mixtures (ACGS) and energetic snacks containing GS (GSS). Data on sequencing were aligned with the reference gene to confirm the PCR products. The regression curve (y = −3.394 x + 42.521; R2 = 0.994, d.f. 14) between Ct values and Log DNA concentrations of Gryllodes sigillatus resulted in an efficiency of 96.4%. The severity of the technological processing treatments and the matrix structure affected the intensity of the PCR signal with the same amount of insect DNA as observed by different y-intercepts of the three-regression lines for RoGS, ACGS, and GSS. The real-time PCR method resulted in robust and sensitive outcomes able to detect low amounts of GS DNA (5 g/100 g) in a complex matrix, making it suitable for detecting the presence or absence of labeled Gryllodes sigillatus material both in feed and food. Keywords Gryllodes sigillatus · Detection · Edible insects · Real-time PCR · Processed food · Feed
Highlights • The method allows for the identification of Gryllodes
sigillatus in a complex matrix.
• Primers exhibited high sensitivity and accuracy. • PCR discrimination among insect species. • The qPCR system was successfully tested in complex
matrices.
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00217-020-03573-1) contains supplementary material, which is available to authorized users. * Francesca Tulli [email protected] 1
Department of Agricultural, Food, Environment and Animal Science, University of Udine, Via Sondrio 2 A, 33100 Udine, Italy
Introduction Global welfare standards and the increasing world population [1] require/necessitate/demand/call for the consideration of protein sources derived from sustainable production systems capable of efficiently converting biomass [2–5]. The novel proteins from unconventional sources should be safe, nutritious, flexible and reliable as well as accepted by consumers. Insects may represent a more efficient way of producing animal protein and seem to meet this goal due to the use of cost-effective raw materials and environmental sustainability along with high nutritional value [6, 7]. Recently, the
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