Novel diagnostic tools for Asian (Apis cerana) and European (Apis mellifera) honey authentication

Abstract

Honey can be produced by different species of honeybees, with two being of economic importance due to their use in apiculture, namely Apis mellifera (known as European honeybee) and Apis cerana (known as Asian honeybee). Due to the decline of the wild populations of the Asian honeybee, this honey generally attains much higher market value, being prone to adulteration. This work aims at proposing new tools, based on the use of molecular markers, for the entomological authentication of honey. To this end, new species-specific primers were designed targeting the tRNA-cox2 intergenic region and allowing the detection of A. cerana DNA by qualitative polymerase chain reaction (PCR). Additionally, a novel real-time PCR method with high resolution melting analysis was developed to target the 16S rRNA gene of both bee species, allowing their discrimination in different clusters. The proposed methodologies were further applied with success in the authentication of Asian and European honey samples by the identification of honeybee DNA, demonstrating the usefulness of these simple and cost-effective new approaches.

Klasifikasi Madu Berdasarkan Jenis Lebah (Apis dorsata versus Apis mellifera) Menggunakan Spektroskopi Ultraviolet dan Kemometrika

“…Selain itu, di Indonesia madu monofloral juga dikelompokkan lagi ke dalam tiga jenis bergantung pada asal lebah yang menghasilkan sekresi madu (asal entomologi), yaitu madu hutan monolforal yang dihasilkan oleh lebah Apis dorsata (MMAD), madu ternak monofloral yang dihasilkan oleh lebah Apis mellifera (MMAM) (yang dikenal juga sebagai lebah Eropa) dan madu ternak monofloral yang dihasilkan oleh lebah Apis cerana (MMAC) (yang dikenal juga sebagai lebah Asia). Secara umum, madu MMAD dan MMAC harganya bisa 310 kali lipat dibandingkan dengan harga madu MMAM (Soares et al 2018;Zhang et al 2019). Perbedaan harga ini terkait dengan keterbatasan jumlah produksi, ketersediaan, dan kesulitan panen madu MMAD dan MMAC.…”

“…yang dioplos dengan pemanis dari molase beras (Sobrino-Gregorio et al 2019). Metode PCR juga telah digunakan untuk membedakan antara madu Apis cerana dan madu Apis mellifera (Soares et al 2018). Metode modern tersebut secara umum memiliki beberapa kelebihan, yaitu persiapan sampel yang minimal, pengukuran yang relatif lebih cepat, dan akurasi pengukuran yang dapat diandalkan.…”

Vibrational Spectroscopy Coupled to a Multivariate Analysis Tiered Approach for Argentinean Honey Provenance Confirmation

“…This technique has been tested also for geographical discrimination purposes, but its application suffers from methodological shortages and limitations [ 3 , 5 ]. Consequently, novel alternative approaches have been proposed, including those based on mass spectrometry, vibrational spectroscopy and molecular biology [ 6 , 7 , 8 ]. The targeted quantification of specific compounds indicative for certain properties and/or origin would represent the most straightforward approach for food authentication; the comparison of the measured parameter with a control limit would empower the direct assessment of the product compliance and might also be used for forensic purposes [ 9 ].…”

Comparison of DNA extraction methods for entomological origin identification of honey using simple additive weighting method

“…() have also successfully used the mitochondrial large subunit ribosomal RNA (16S rRNA) gene, while Soares et al . () used mitochondrial tRNAleu‐cox2 (cytochrome c oxidase subunit II) intergenic region and the 16S rRNA gene for bee identification in honey samples. The 16S rRNA gene is one of the commonly used mitochondrial DNA (mtDNA) in bees' phylogenetic relationship studies (Cameron et al ., ; Raffiudin & Crozier, ; Rasmussen & Cameron, ).…”

“…Prosser & Hebert (2017) have identified bee source of Apis mellifera, a common European honey bee and also the stingless bee of Melipona beecheii in honey using the COI gene. Kek et al (2017b) have also successfully used the mitochondrial large subunit ribosomal RNA (16S rRNA) gene, while Soares et al (2018) used mitochondrial tRNAleu-cox2 (cytochrome c oxidase subunit II) intergenic region and the 16S rRNA gene for bee identification in honey samples. The 16S rRNA gene is one of the commonly used mitochondrial DNA (mtDNA) in bees' phylogenetic relationship studies (Cameron et al, 1992;Raffiudin & Crozier, 2007;Rasmussen & Cameron, 2010).…”