COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European Honey

Honey is a widely consumed food product frequently subjected to adulteration, with the mislabeling of its botanical or geographical origin being one of the most common practices. Determining the entomological origin of honey is particularly challenging but of high relevance for ensuring its authenti...

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Main Authors: Mónica Honrado, Andreia Quaresma, Dora Henriques, M. Alice Pinto, Joana S. Amaral
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Foods
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Online Access:https://www.mdpi.com/2304-8158/14/3/419
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author Mónica Honrado
Andreia Quaresma
Dora Henriques
M. Alice Pinto
Joana S. Amaral
author_facet Mónica Honrado
Andreia Quaresma
Dora Henriques
M. Alice Pinto
Joana S. Amaral
author_sort Mónica Honrado
collection DOAJ
description Honey is a widely consumed food product frequently subjected to adulteration, with the mislabeling of its botanical or geographical origin being one of the most common practices. Determining the entomological origin of honey is particularly challenging but of high relevance for ensuring its authenticity, especially for products with protected designation of origin (PDO) labels. This study presents a novel DNA metabarcoding approach targeting a highly informative 406 bp fragment of the cytochrome c oxidase I (COI) gene to differentiate among the three major mitochondrial lineages (A, M, and C) of honey bees (<i>Apis mellifera</i> L.) native to Europe. The target region was selected based on the calculated fixation index (F<sub>ST</sub>), which is frequently used in Population Genetics as a measure of differentiation between populations. The approach was validated with 11 honey samples of known entomological origin and applied to 44 commercial honeys from 13 countries. The approach demonstrated high sensitivity, accurately identifying the entomological origin of honey, including samples produced by honey bees of varying ancestries, which could not be resolved by previous methods based on real-time PCR coupled with high-resolution melting (PCR-HRM) analysis. The results demonstrate the effectiveness of COI metabarcoding in verifying honey authenticity and highlight the predominance of C-lineage honey bees in the production of commercial honeys from northwestern Europe. This finding suggests a limited presence of the native M-lineage ancestry, underscoring the need for conservation efforts.
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spelling doaj-art-af39efc99a3141f38ffd9cbb677428972025-08-20T02:48:09ZengMDPI AGFoods2304-81582025-01-0114341910.3390/foods14030419COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European HoneyMónica Honrado0Andreia Quaresma1Dora Henriques2M. Alice Pinto3Joana S. Amaral4CIMO, LA SusTEC, Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253 Bragança, PortugalCIMO, LA SusTEC, Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253 Bragança, PortugalCIMO, LA SusTEC, Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253 Bragança, PortugalCIMO, LA SusTEC, Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253 Bragança, PortugalCIMO, LA SusTEC, Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253 Bragança, PortugalHoney is a widely consumed food product frequently subjected to adulteration, with the mislabeling of its botanical or geographical origin being one of the most common practices. Determining the entomological origin of honey is particularly challenging but of high relevance for ensuring its authenticity, especially for products with protected designation of origin (PDO) labels. This study presents a novel DNA metabarcoding approach targeting a highly informative 406 bp fragment of the cytochrome c oxidase I (COI) gene to differentiate among the three major mitochondrial lineages (A, M, and C) of honey bees (<i>Apis mellifera</i> L.) native to Europe. The target region was selected based on the calculated fixation index (F<sub>ST</sub>), which is frequently used in Population Genetics as a measure of differentiation between populations. The approach was validated with 11 honey samples of known entomological origin and applied to 44 commercial honeys from 13 countries. The approach demonstrated high sensitivity, accurately identifying the entomological origin of honey, including samples produced by honey bees of varying ancestries, which could not be resolved by previous methods based on real-time PCR coupled with high-resolution melting (PCR-HRM) analysis. The results demonstrate the effectiveness of COI metabarcoding in verifying honey authenticity and highlight the predominance of C-lineage honey bees in the production of commercial honeys from northwestern Europe. This finding suggests a limited presence of the native M-lineage ancestry, underscoring the need for conservation efforts.https://www.mdpi.com/2304-8158/14/3/419entomologic origin<i>Apis mellifera</i>mtDNADNA metabarcodinghoneyCOI gene
spellingShingle Mónica Honrado
Andreia Quaresma
Dora Henriques
M. Alice Pinto
Joana S. Amaral
COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European Honey
Foods
entomologic origin
<i>Apis mellifera</i>
mtDNA
DNA metabarcoding
honey
COI gene
title COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European Honey
title_full COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European Honey
title_fullStr COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European Honey
title_full_unstemmed COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European Honey
title_short COI Metabarcoding as a Novel Approach for Assessing the Honey Bee Source of European Honey
title_sort coi metabarcoding as a novel approach for assessing the honey bee source of european honey
topic entomologic origin
<i>Apis mellifera</i>
mtDNA
DNA metabarcoding
honey
COI gene
url https://www.mdpi.com/2304-8158/14/3/419
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