Novel real-time PCR protocol for the detection of house cricket (Acheta domesticus) in feed
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2021
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Prikaz svih podataka o dokumentuApstrakt
A new real-time PCR protocol was developed for the detection of house cricket (Acheta domesticus) in feeds. A sequence of 97 bp size within the mitochondrial barcode region (cytochrome oxidase c subunit I - COI) was amplified by the chosen specific primer and probe set. The specificity of the PCR system was confirmed with a broad range of plant and animal species. The limit of detection with pure A. domesticus DNA diluted in aquaculture feed DNA was 1 genome copy, corresponding to an absolute amount of 2.14 pg DNA. In addition, the sensitivity was investigated in dependence on different processing treatments of the insect material. The method reliably detected pasteurized (80 degrees C, 10 min) and oven-dried (60 degrees C, 3 h) A. domesticus in feed mixtures down to 0.1% (w/w). Practical applicability of the proposed protocol was tested on 33 commercial feeds not declaring the presence of house cricket in the list of ingredients, and on two model feeds spiked in-house with A. domestic...us. The method clearly recognized the presence of house cricket in model feeds down to 0.1%, a concentration residing in the range of accidental contamination rather than deliberate addition. It is concluded that the tested system is suitable for practical application, enabling the sensitive and reliable detection of A. domesticus material as a newly allowed component in aquaculture feed.
Ključne reči:
Real-time PCR / Insect / Aquaculture feed / Acheta domesticusIzvor:
Animal Feed Science and Technology, 2021, 280, 115057-Izdavač:
- Amsterdam : Elsevier
Finansiranje / projekti:
- Guest scientist program (Werner-Baltes-Fellowship) of the German Federal Institute for Risk Assessment (BfR)
- Unapređenje i razvoj higijenskih i tehnoloških postupaka u proizvodnji namirnica životinjskog porekla u cilju dobijanja kvalitetnih i bezbednih proizvoda konkurentnih na svetskom tržištu (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-46009)
- EU funded EFSA initiative to promote the training of new risk assessors in the EU
- Federal Ministry of Food and Agriculture (BMEL)
- national project "Allergen-Pro" [281A304A18]
DOI: 10.1016/j.anifeedsci.2021.115057
ISSN: 0377-8401
WoS: 000703431100011
Scopus: 2-s2.0-85114416405
Institucija/grupa
Naučni institut za veterinarstvo SrbijeTY - JOUR AU - Garino, Cristiano AU - Zagon, Jutta AU - Nešić, Ksenija PY - 2021 UR - https://reponivs.nivs.rs/handle/123456789/454 AB - A new real-time PCR protocol was developed for the detection of house cricket (Acheta domesticus) in feeds. A sequence of 97 bp size within the mitochondrial barcode region (cytochrome oxidase c subunit I - COI) was amplified by the chosen specific primer and probe set. The specificity of the PCR system was confirmed with a broad range of plant and animal species. The limit of detection with pure A. domesticus DNA diluted in aquaculture feed DNA was 1 genome copy, corresponding to an absolute amount of 2.14 pg DNA. In addition, the sensitivity was investigated in dependence on different processing treatments of the insect material. The method reliably detected pasteurized (80 degrees C, 10 min) and oven-dried (60 degrees C, 3 h) A. domesticus in feed mixtures down to 0.1% (w/w). Practical applicability of the proposed protocol was tested on 33 commercial feeds not declaring the presence of house cricket in the list of ingredients, and on two model feeds spiked in-house with A. domesticus. The method clearly recognized the presence of house cricket in model feeds down to 0.1%, a concentration residing in the range of accidental contamination rather than deliberate addition. It is concluded that the tested system is suitable for practical application, enabling the sensitive and reliable detection of A. domesticus material as a newly allowed component in aquaculture feed. PB - Amsterdam : Elsevier T2 - Animal Feed Science and Technology T1 - Novel real-time PCR protocol for the detection of house cricket (Acheta domesticus) in feed SP - 115057 VL - 280 DO - 10.1016/j.anifeedsci.2021.115057 UR - conv_572 ER -
@article{ author = "Garino, Cristiano and Zagon, Jutta and Nešić, Ksenija", year = "2021", abstract = "A new real-time PCR protocol was developed for the detection of house cricket (Acheta domesticus) in feeds. A sequence of 97 bp size within the mitochondrial barcode region (cytochrome oxidase c subunit I - COI) was amplified by the chosen specific primer and probe set. The specificity of the PCR system was confirmed with a broad range of plant and animal species. The limit of detection with pure A. domesticus DNA diluted in aquaculture feed DNA was 1 genome copy, corresponding to an absolute amount of 2.14 pg DNA. In addition, the sensitivity was investigated in dependence on different processing treatments of the insect material. The method reliably detected pasteurized (80 degrees C, 10 min) and oven-dried (60 degrees C, 3 h) A. domesticus in feed mixtures down to 0.1% (w/w). Practical applicability of the proposed protocol was tested on 33 commercial feeds not declaring the presence of house cricket in the list of ingredients, and on two model feeds spiked in-house with A. domesticus. The method clearly recognized the presence of house cricket in model feeds down to 0.1%, a concentration residing in the range of accidental contamination rather than deliberate addition. It is concluded that the tested system is suitable for practical application, enabling the sensitive and reliable detection of A. domesticus material as a newly allowed component in aquaculture feed.", publisher = "Amsterdam : Elsevier", journal = "Animal Feed Science and Technology", title = "Novel real-time PCR protocol for the detection of house cricket (Acheta domesticus) in feed", pages = "115057", volume = "280", doi = "10.1016/j.anifeedsci.2021.115057", url = "conv_572" }
Garino, C., Zagon, J.,& Nešić, K.. (2021). Novel real-time PCR protocol for the detection of house cricket (Acheta domesticus) in feed. in Animal Feed Science and Technology Amsterdam : Elsevier., 280, 115057. https://doi.org/10.1016/j.anifeedsci.2021.115057 conv_572
Garino C, Zagon J, Nešić K. Novel real-time PCR protocol for the detection of house cricket (Acheta domesticus) in feed. in Animal Feed Science and Technology. 2021;280:115057. doi:10.1016/j.anifeedsci.2021.115057 conv_572 .
Garino, Cristiano, Zagon, Jutta, Nešić, Ksenija, "Novel real-time PCR protocol for the detection of house cricket (Acheta domesticus) in feed" in Animal Feed Science and Technology, 280 (2021):115057, https://doi.org/10.1016/j.anifeedsci.2021.115057 ., conv_572 .