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Published: 13 May 2025
Detection of Bombyx mori as a Protein Source in Feedingstuffs by Real-Time PCR with a Single-Copy Gene Target. Agriculture, 14(11), 1996
Authors:
Marien, A., Dubois, B., Anselmo, A., Veys, P., Berben, G., Kohl, C., Maljean, J., Guillet, S., Morin, J., & Debode, F.
Abstract:
The silkworm, Bombyx mori, is reared on a large scale, mainly for silk production. The waste from this silk production, like pupae, is underused. As an edible insect, B. mori 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 (B. mori). 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 B. mori 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.Keywords:insect; Bombyx mori; silkworm; lepidoptera; detection; real-time PCR; cadherin; feed
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Published: 13 May 2025
Detection of Acheta domesticus by real-time PCR in food and feed. Journal of Insects as Food and Feed, 10(9), 1645–1660
Authors:
Jilkova, D., Marien, A., Hulin, J., Zdenkova, K., Fumiere, O., Cermakova, E., Berben, G., & Debode, F.
Abstract:
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.
https://doi.org/10.1163/23524588-00001067
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Published: 13 May 2025
Detection of Alphitobius diaperinus by Real-Time Polymerase Chain Reaction With a Single-Copy Gene Target. Frontiers in Veterinary Science, 9
Authors:
Marien, A., Sedefoglu, H., Dubois, B., Maljean, J., Francis, F., Berben, G., Guillet, S., Morin, J., Fumière, O., & Debode, F.
Abstract:
Use of edible insects as an alternative source of proteins in food and feed is increasing. These last years, numerous companies in Europe have started producing insects for food and feed purposes. In the European Union, the use of edible insects for human consumption falls within Regulation (EU) No. 2015/2283 on novel foods. For feed, Commission Regulation (EU) 2017/893 authorizes seven insect species as processed animal proteins for aquaculture. Methods of authentication are required to check the conformity of the products. In this study, we propose a real-time polymerase chain reaction (PCR) method for the specific detection of the lesser mealworm (Alphitobius diaperinus), one of the species included in the shortlist of authorized insects. The selected target is the cadherin gene with a single-copy (per haploid genome) illustrated by our experimental evidence. The PCR test amplified a 134-bp fragment of the cadherin gene. The qualitative method was assessed toward several performance criteria. Specificity was checked against 54 insect species next to other animal and plant species. The sensitivity, efficiency, robustness, and transferability of the PCR assay were also successfully tested. Finally, the applicability of the test was assessed on real-life processed samples (industrial meals) of A. diaperinus. The study also showed that there seems to be a huge confusion on the correct labeling of the marketed mealworms. We did not succeed to get Alphitobius laevigatus samples. They all appeared to belong to the A. diaperinus taxon.
https://doi.org/10.3389/fvets.2022.718806
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Published: 13 May 2025
Chromosome-scale assembly of the yellow mealworm genome - Open Research Europe, 1, 94
Authors:
Eleftheriou, E., Aury, J., Vacherie, B., Istace, B., Belser, C., Noel, B., Moret, Y., Rigaud, T., Berro, F., Gasparian, S., Labadie-Bretheau, K., Lefebvre, T., & Madoui, M.
Abstract:
Background: The yellow mealworm beetle, Tenebrio molitor, is a promising alternative protein source for animal and human nutrition and its farming involves relatively low environmental costs. For these reasons, its industrial scale production started this century. However, to optimize and breed sustainable new T. molitor lines, the access to its genome remains essential.
Methods: By combining Oxford Nanopore and Illumina Hi-C data, we constructed a high-quality chromosome-scale assembly of T. molitor. Then, we combined RNA-seq data and available coleoptera proteomes for gene prediction with GMOVE.
Results: We produced a high-quality genome with a N50 = 21.9Mb with a completeness of 99.5% and predicted 21,435 genes with a median size of 1,780 bp. Gene orthology between T. molitor and Tribolium castaneum showed a highly conserved synteny between the two coleoptera and paralogs search revealed an expansion of histones in the T. molitor genome.
Conclusions: The present genome will greatly help fundamental and applied research such as genetic breeding and will contribute to the sustainable production of the yellow mealworm.
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