Authentication of honey based on a DNA method to differentiate Apis mellifera subspecies: Application to Sicilian honey bee ( A. m. siciliana ) and Iberian honey bee ( A. m. iberiensis ) honeys

Honey as a Source of Environmental DNA for the Detection and Monitoring of Honey Bee Pathogens and Parasites

“…DNA extracted from all honey samples was amplified for the Apis mellifera mtDNA targeted region and the obtained fragments were the expected ones [ 44 ]. That means that for all samples, DNA was successfully isolated and PCR amplification was not inhibited by any contaminants eventually left in the extracted DNA aliquots (spectrophotometer analyses confirmed this result: A 260 /A 280 was always >1.6).…”

“…Honey is a complex matrix that is mainly made by sugars but that contains several other components that can be explored to disclose interesting information that could be used for several purposes. One of the components, usually neglected or not considered at all, is the DNA that derives from all organisms that directly or indirectly contributed to its productions or that were part of the production systems or environments where the honey has been produced [ 6 , 7 , 43 , 44 , 45 ]. This eDNA also contains information on pathogens and parasites that can be present in the beehive ecological niche and that can represent a threat for the honey bee health.…”

“…DNA from the honey bees is always contained in the honey they produce [ 44 ]. Therefore, to evaluate the quality of the extracted DNA and the possibility to use it for other tests, we first amplified the extracted nucleic acid with primers that targeted a region of the honey bee mtDNA that can be also used to detect the mitochondrial lineage of the A. mellifera that produced the honey [ 44 ].…”

“…PCR analyses were carried out using the PCR primer pairs reported in Table 1 . To assess the possibility to successfully amplify DNA fragments from the extracted DNA, we first verified if amplification could occur for honey bee DNA using primers designed on the mitochondrial DNA (mtDNA) region [ 44 ]. Primer pairs used in this study that targeted sequences of Paenibacillus larvae (two primer pairs) Melissococcus plutonius (two primer pairs) Nosema apis (one primer pair), N. ceranae (one primer pair), Ascosphaera apis (one primer pair), Lotmaria passim (one primer pair), Acarapis woodi (two primer pairs), and Varroa destructor (two primer pairs) were described by other authors ( Table 1 ).…”

An Alternative, High Throughput Method to Identify Csd Alleles of the Honey Bee

“…Diagnostic methods used in honey analysis for food control imply DNA-based techniques, focusing on the botanical origin of the honey [22,23] and the identification of its entomological origin via molecular markers [24][25][26][27][28]. According to our knowledge, this approach has not been used for tracking the genetic composition of a mating yard thus far.…”