Browsing by Author "Medibees Consortium"
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- Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeysPublication . Honrado, Mónica; Henriques, Dora; Santos, Joana; Yadró Garcia, Carlos A.; Medibees Consortium; Pinto, M. Alice; Amaral, Joana S.As defined by the Codex Alimentarius, honey is the natural sweet substance produced by honeybees from the nectar of plants.1 This natural product is widely appreciated but is also considered one of the foods most prone to adulteration. The increasing demand for monofloral honey and those with protected designation of origin (PDO) has led to increased fraud by mislabeling botanical and geographical origin.2 Verifying the geographical origin of honey is a challenging endeavor. Recently, attention has been paid to the entomological origin, as it aligns with the geographical patterns of honeybee subspecies. The Mediterranean region is a hot spot of Apis mellifera subspecific diversity shaped by thousands of years of evolution. Although contemporary human-mediated movements of queens have impacted the native subspecific distribution, several PDO honeys specify the subspecies that produce those honeys, thus offering a unique avenue for authentication. As part of the European PRIMA project MEDIBEES, we aim to develop a DNA-metabarcoding approach to authenticate honey's entomological origin, focusing on mitochondrial lineages A, M, C, and O. To achieve this, the DNA of 1280 honeybees representing 16 subspecies and the four lineages (A.m. sahariensis, A.m. intermisa, A.m. siciliana, A.m. ruttneri, A.m. iberiensis, A.m. ligustica, A.m. macedonica, A.m. adami, A.m. cecropia, A.m. cypria, A.m. caucasica, A.m. meda, A.m. anatoliaca, A.m. syriaca, A.m. jemenitica, A.m. lamarcki) was extracted, and their whole genomes were sequenced. The MitoZ software was used to assemble the mitochondrial genomes, resulting in 769 mitochondrial genomes successfully assembled. Subsequently, each of these genomes was aligned individually with a reference genome using MEGA software, and mitogenomes not specific to Apis mellifera were discarded. Of these, only the mitogenomes corresponding to the native ancestry were retained, resulting in a final set of 355 mitogenomes in the database. A phylogenetic analysis was conducted with the final 355 mitochondrial sequences, revealing four distinct clusters corresponding to the four maternal lineages. This dataset was used for calculating the fixation index (FST) pairwise values, and a sliding window of 400 bp was used to identify single nucleotide polymorphisms (SNPs) that effectively differentiate (FST>0.98) the four lineages, enabling the identification of promising regions for primer design. This work resulted in the discovery of three promising regions for discriminating the four maternal lineages: one in the COI gene, one in the ND1 gene, and one in the CYTB gene (Fig. 1).
- General beekeeping practices and main stressors identified by beekeepers in the MediterraneanPublication . Nanetti, Antonio; Martín-Hernández, Raquel; Zammit Mangion, Marion; Alice Pinto, M.; Farrugia, Dylan; Zammit Mangion, Rachel; Noureddine, Adjlane; Eissa, Asmaa Anwar; Cilia, Giovanni; Daour, Ahmad Yousef; Garrido, Claudia; Haddad, Nizar; Henriques, Dora; Hosri, Chadi; Muz, Mustafa Necati; Sagastume, Soledad; Galea, Thomas; Medibees ConsortiumBeekeeping is an ecologically and economically important activity in the Mediterranean that is increasingly under threat from a combination of factors. In the MEDIBEES (Monitoring the Mediterranean honey bee subspecies and their resilience to climate change for the improvement of sustainable agro-ecosystems) PRIMA project we aim to identify honey bees that show resilience to stressors associated with climate change. An important first step in our work was to establish baseline data of practices in the region through the administering of a questionnaire on the beekeeping practices. Over 1022 responses were received from beekeepers in Algeria, Egypt, Jordan, Italy, Lebanon, Malta, Spain, Portugal and Turkey. Beekeeping was overwhelmingly male dominated (ratio 9:1) and practiced largely by the middle-aged (mainly between 40-50 years). The majority of beekeepers reported having 10-50 boxes with the exceptions of Algeria and Jordan where the majority reported keeping more than 100 hive boxes. Across the Mediterranean the Langstroth hive box was most commonly used. Despite the perception of migratory beekeeping being frequently practiced, only Lebanon and Turkey reported significant levels of transhumance (>50.0 and 70.0% of beekeepers respectively). 51.2% of beekeepers reported practicing queen rearing with the majority of these (51.4%) reporting re-queening their colonies every 2 years. Interestingly, the majority of all beekeepers agreed with the statement that their native honey bee is endangered where pesticide use, lack of adequate forage and parasite infestations such as the Varroa mite were repeatedly cited as causing the greatest losses to honey bees in general. Climate change and urbanisation were also linked with colony losses by causing habitat loss. Supplementary feeding was reported to be an integral part of beekeeping in this region, with beekeepers reporting using between 0-5kg, closely followed by 6-10 kg of additional sugars.
- MEDIBEES: Monitoring the Mediterranean honey bee subspecies and their resilience to climate change for the improvement of sustainable agro-ecosystemsPublication . Martín-Hernández, Raquel; Nanetti, Antonio; Pinto, M. Alice; Noureddine, Adjlane; Haddad, Nizar; Hosri, Chadi; Muz, Mustafa Necati; Zammit Mangion, Marion; Daour, Ahmad Yousef; Sagastume, Soledad; Cilia, Giovanni; Medibees ConsortiumBeekeeping provides livelihood for hundreds of thousands of beekeepers in the Mediterranean area. This activity relies on a number of different indigenous Apis mellifera subspecies, adapted to the very diverse and harsh conditions of the region. Climate change is expected to increase the stress factors affecting bees, especially in this region, reducing both pollination efficiency and production potential. Unfortunately, our ability to address this problem is limited by the incomplete knowledge of the natural adaptation mechanisms developed by the different subspecies. In order to increase the knowledge base for future selection programs to improve bee populations for environmental changes, the MEDIBEES project is being developed. It includes 9 partners from 8 Mediterranean countries on all three shores of the Mediterranean, covering 10 local A. mellifera subspecies, which represents a remarkable though understudied proportion of the species genetic diversity. The project aims to: a) unravel the differential genetic background of Mediterranean subspecies, b) understand their adaptation to local conditions, and c) characterize their resistance to climate change. To achieve the objectives, colonies belonging to the local subspecies will be studied phenotypically to determine their behavior under environmental conditions, covering survival, sensitivity to pests/pathogens, behavior, physiology and reproduction which will be completed by gene expression and transcriptomic assays. In addition, complete genomes of field and laboratory samples will be sequenced to find genes putatively involved in adaptation and to develop new genetic tools to characterize honey bee populations according to their resistance to environmental stress factors. This effort will encourage the use of local subspecies, to make them more attractive and avoid importing foreign breeds, and will lay the foundation for future selection programs. Besides, the valorization of honey by both promoting its use and developing quality labels, and the evaluation of beekeeping by-products as modifiers of soil fertility and biota are also approached to help the beekeepers improve the sustainability of their farms in an economical and environmental sound manner.
- A sequenciação de nova geração como uma abordagem promissora para a identificação da origem entomológica do melPublication . Honrado, Mónica; Henriques, Dora; Yadró Garcia, Carlos A.; Santos, Joana; Rufino, José; Medibees Consortium; Pinto, M. Alice; Amaral, Joana S.O mel é um alimento muito consumido e apreciado em todo o mundo pelas suas propriedades nutricionais e organoléticas, bem como pelos seus efeitos benéficos para a saúde. No entanto, é também considerado um dos alimentos mais suscetíveis de ser adulterado, quer pela mistura de mel de qualidade inferior, quer pela adição de açúcares, ou pela rotulagem incorreta da origem botânica e/ou geográfica, entre outras possíveis fraudes. Nos últimos anos, tem sido dada uma atenção crescente à origem entomológica do mel, uma vez que esta também está relacionada com a origem geográfica. No âmbito do projeto PRIMA “MEDIBEES” (https://medibees.org/), a sequenciação de nova geração (NGS) será utilizada com vista ao desenvolvimento de ferramentas moleculares que permitam identificar a origem entomológica de amostras de mel provenientes dos 8 países mediterrânicos do consórcio, de forma a diferenciar e valorizar méis produzidos por abelhas autóctones destes países. Com este objetivo, inicialmente procedeu-se à construção da base de dados das sequências de DNA mitocondrial das abelhas de modo a incluir 10 subespécies mediterrânicas das 4 linhagens maternas (A, M, C e O). Para tal, procedeu-se à extração de DNA e à respetiva sequenciação dos genomas completos, na plataforma Illumina Novaseq 6000, de um total de 1095 abelhas destes países. Posteriormente, utilizou-se o programa mitoZ 3.6 para fazer a montagem do genoma mitocondrial de cada uma das amostras, resultando na seleção de 283 sequências mitocondriais com boa montagem. Em seguida, foi utilizado o software MEGA 11, para realizar o alinhamento destas sequências. A informação obtida será posteriormente utilizada para a seleção de regiões com variantes (SNPs) informativos que possam ser usadas para o desenho de primers adequados e desenvolvimento de ferramentas para a identificação de méis produzidos por abelhas de diferentes linhagens mitocondriais e respetivas subespécies.