Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target

The silkworm, <i>Bombyx mori</i>, is reared on a large scale, mainly for silk production. The waste from this silk production, like pupae, is underused. As an edible insect, <i>B. mori</i> is a good source of protein in human food and animal feed. In recent years, European le...

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Main Authors: Aline Marien, Benjamin Dubois, Abigaël Anselmo, Pascal Veys, Gilbert Berben, Cloé Kohl, Julien Maljean, Stéphanie Guillet, Jean-François Morin, Frédéric Debode
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Agriculture
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Online Access:https://www.mdpi.com/2077-0472/14/11/1996
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author Aline Marien
Benjamin Dubois
Abigaël Anselmo
Pascal Veys
Gilbert Berben
Cloé Kohl
Julien Maljean
Stéphanie Guillet
Jean-François Morin
Frédéric Debode
author_facet Aline Marien
Benjamin Dubois
Abigaël Anselmo
Pascal Veys
Gilbert Berben
Cloé Kohl
Julien Maljean
Stéphanie Guillet
Jean-François Morin
Frédéric Debode
author_sort Aline Marien
collection DOAJ
description The silkworm, <i>Bombyx mori</i>, is reared on a large scale, mainly for silk production. The waste from this silk production, like pupae, is underused. As an edible insect, <i>B. mori</i> is a good source of protein in human food and animal feed. In recent years, European legislation on the use of insects has evolved and a multitude of European companies have initiated the rearing of insects specifically for food and feed applications. Regarding animal feed, Commission Regulations (EU) 2021/1372 and 2021/1925 authorize eight insect species, including silkworm, as processed animal proteins for use in fish, pig, and poultry feed. The incorporation of edible insects into the human diet falls within Regulation (EU) No. 2015/2283 concerning novel foods. Implementation of authentication methods is imperative to ensure the conformity of the products. In the present study, we propose a specific real-time PCR method for the detection of silkworm (<i>B. mori</i>). The developed PCR test amplifies a 98 bp fragment of the cadherin gene. This gene is present in a single-copy per haploid genome, as demonstrated by experimental evidence. The qualitative method was successfully evaluated on the performance criteria of specificity, sensitivity, efficiency, robustness, and transferability. The applicability of the test was assessed on samples of <i>B. mori</i> from industry. Light microscopy and DNA metabarcoding approaches were used as a complement to genomic analysis as a means of providing authentication of the samples.
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spelling doaj-art-fd40ff41bf534de69a412724cec8d5c02025-08-20T02:26:51ZengMDPI AGAgriculture2077-04722024-11-011411199610.3390/agriculture14111996Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene TargetAline Marien0Benjamin Dubois1Abigaël Anselmo2Pascal Veys3Gilbert Berben4Cloé Kohl5Julien Maljean6Stéphanie Guillet7Jean-François Morin8Frédéric Debode9Quality and Authentication of Agricultural Products Unit, Knowledge and Valorization of agricultural Products Department, Walloon Agricultural Research Center (CRA-W), 5030 Gembloux, BelgiumBioengineering Unit, Life Sciences Department, Walloon Agricultural Research Center (CRA-W), 5030 Gembloux, BelgiumQuality and Authentication of Agricultural Products Unit, Knowledge and Valorization of agricultural Products Department, Walloon Agricultural Research Center (CRA-W), 5030 Gembloux, BelgiumQuality and Authentication of Agricultural Products Unit, Knowledge and Valorization of agricultural Products Department, Walloon Agricultural Research Center (CRA-W), 5030 Gembloux, BelgiumQuality and Authentication of Agricultural Products Unit, Knowledge and Valorization of agricultural Products Department, Walloon Agricultural Research Center (CRA-W), 5030 Gembloux, BelgiumHaute Ecole de la Province de Liège (HEPL), 4000 Liège, BelgiumQuality and Authentication of Agricultural Products Unit, Knowledge and Valorization of agricultural Products Department, Walloon Agricultural Research Center (CRA-W), 5030 Gembloux, BelgiumEurofins Biologie Moléculaire France, Eurofins, 44323 Nantes, FranceEurofins Biologie Moléculaire France, Eurofins, 44323 Nantes, FranceBioengineering Unit, Life Sciences Department, Walloon Agricultural Research Center (CRA-W), 5030 Gembloux, BelgiumThe silkworm, <i>Bombyx mori</i>, is reared on a large scale, mainly for silk production. The waste from this silk production, like pupae, is underused. As an edible insect, <i>B. mori</i> is a good source of protein in human food and animal feed. In recent years, European legislation on the use of insects has evolved and a multitude of European companies have initiated the rearing of insects specifically for food and feed applications. Regarding animal feed, Commission Regulations (EU) 2021/1372 and 2021/1925 authorize eight insect species, including silkworm, as processed animal proteins for use in fish, pig, and poultry feed. The incorporation of edible insects into the human diet falls within Regulation (EU) No. 2015/2283 concerning novel foods. Implementation of authentication methods is imperative to ensure the conformity of the products. In the present study, we propose a specific real-time PCR method for the detection of silkworm (<i>B. mori</i>). The developed PCR test amplifies a 98 bp fragment of the cadherin gene. This gene is present in a single-copy per haploid genome, as demonstrated by experimental evidence. The qualitative method was successfully evaluated on the performance criteria of specificity, sensitivity, efficiency, robustness, and transferability. The applicability of the test was assessed on samples of <i>B. mori</i> from industry. Light microscopy and DNA metabarcoding approaches were used as a complement to genomic analysis as a means of providing authentication of the samples.https://www.mdpi.com/2077-0472/14/11/1996insect<i>Bombyx mori</i>silkwormlepidopteradetectionreal-time PCR
spellingShingle Aline Marien
Benjamin Dubois
Abigaël Anselmo
Pascal Veys
Gilbert Berben
Cloé Kohl
Julien Maljean
Stéphanie Guillet
Jean-François Morin
Frédéric Debode
Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target
Agriculture
insect
<i>Bombyx mori</i>
silkworm
lepidoptera
detection
real-time PCR
title Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target
title_full Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target
title_fullStr Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target
title_full_unstemmed Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target
title_short Detection of <i>Bombyx mori</i> as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target
title_sort detection of i bombyx mori i as a protein source in feedingstuffs by real time pcr with a single copy gene target
topic insect
<i>Bombyx mori</i>
silkworm
lepidoptera
detection
real-time PCR
url https://www.mdpi.com/2077-0472/14/11/1996
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