CIMO - Resumos em Proceedings Não Indexados à WoS/Scopus
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- Resposta do cânhamo (Canabis sativa) à densidade de sementeira e fertilização azotadaPublication . Eje, Röndahl; Arrobas, Margarida; Queijo, Luís; Rocha, João; Rodrigues, Manuel ÂngeloO cânhamo já foi uma cultura muito importante. Durante a época dos Descobrimentos os navios utilizavam até cem toneladas de cordas e têxteis para as velas feitas de cânhamo. Recentemente foram identificados cerca de vinte e cinco mil produtos manufaturados com cânhamo. Os objetivos deste trabalho foram estudar a resposta da densidade da plantação e da fertilização azotada na produtividade de fibra e de semente na cultura do cânhamo. Os ensaios foram conduzidos com irrigação em dois locais no nordeste de Portugal, em Bragança (na Terra Fria) e em Mirandela (na Terra Quente). A cultura foi semeada em três densidades (25, 50 e 100 kg ha-1) e adubada com três doses diferentes de azoto (50, 100 e 200 kg ha-1). Foi também incluída uma modalidade testemunha, sem fertilização azotada, e três repetições de cada tratamento. Os resultados mostraram que ambos os fatores em estudo, densidade de plantação e adubação azotada, tiveram influência na produtividade de uma forma não linear. A maior produção de fibra neste ensaio foi obtida com a cultura semeada na densidade de 25 e 50 kg ha-1 de semente e com uma adubação de 100 kg N ha-1. Com uma adubação superior a 100 kg N ha-1 verifica-se uma diminuição na produtividade que poderá ser justificada por desequilíbrios nutricionais associados ao excesso de azoto. A maior produção de semente neste ensaio foi obtida nos tratamentos com a menor densidade de plantação e nas três menores dosagens de azoto.
- Old and new climate change adaptation strategies by Montesinho mountain pastoralists, PortugalPublication . Aleixo-Pais, Isa G.; Castro, José; Frazão-Moreira, Amélia; Castro, João Paulo; Castro, MarinaMountain pastoralists are among the communities most affected by climate change, rendering empowerment of mountain communities and supportive policies essential.
- Old and new climate change adaptation strategies by Montesinho mountain pastoralists, PortugalPublication . Aleixo-Pais, Isa G.; Castro, José; Frazão-Moreira, Amélia; Castro, João PauloMountain pastoralists are among the communities most affected by climate change, rendering empowerment of mountain communities and supportive policies essential.
- Lake Sinai virus in Azorean Honey bees: Understanding the impact of Varroa destructor on prevalence, loads, and strain distributionPublication . Lopes, Ana; Low, Matthew; Martín-Hernández, Raquel; Miranda, Joachim; Pinto, M. AliceLake Sinai virus (LSV) is a pathogen affecting honey bees worldwide. It was first discovered in 2009 in Lake Sinai, USA, and manifests as a multi-strain virus. The Azores archipelago comprises islands with and without Varroa destructor, offering a unique setting for studying viral epidemiology. Building upon insights from the heterogeneous distribution of Deformed wing virus (DWV) strains, which was modulated by varroa’s invasion, this study sought to evaluate prevalence, load, and diversity of LSV in the Azores, and explore potential changes in the viral landscape attributable to the mite’s presence. In July/August of 2014/15 and 2020, 494 colonies were sampled across the archipelago. These were screened for LSV using RT-qPCR with a primer pair that allowed detection of at least four strains (LSV-1,-2,-3, and -4). Positive samples were further examined by highthroughput sequencing (HTS). The impact of varroa on prevalence and loads was evaluated using general linear mixed models in the framework of Bayesian analysis. LSV was detected on all islands (prevalence range: 7.7%–89.9%), with varroa’s presence significantly increasing prevalence (mean increase: 19.5±9.5, Probability of increase=97.6%). HTS identified two known strains (LSV-2, LSV-3) and one novel strain (LSV-9). Phylogenetic reconstruction revealed a strong geographic structure in which LSV-2 was found dominating all varroa-infested islands (Flores, Faial, and Pico) and one varroa-free island (São Jorge). LSV-3 and LSV-9 dominated varroa-free islands, Graciosa and Terceira, and São Miguel and Santa Maria, respectively. Viral loads varied greatly among islands, from 4.77 log10 copies/bee to 8.71 log10 copies/bee. Varroa’s presence affected LSV-2 loads, with a mean increase of 2.5±0.7 log10 copies/bee (Pr=100%). Our Żndings highlight mitedriven evolutionary changes in LSV in the Azores and identify a novel strain dominating the easternmost islands, coinciding with the unique refuge of DWV-C existing in this part of the archipelago.
- Phenology of the Microsporidia Nosema ceranae in four mediterranean countriesPublication . Jabal-Uriel, Clara; Bonjour-Dalmon, A.; Chejanovsky, Nor; Higes, Mariano; Le Conte, Yves; Lopes, Ana; Pinto, M. Alice; Reyes-Carreño, Maritza; Soroker, Victoria; Martín-Hernández, RaquelThe presence and prevalence of the Microsporidia Nosema ceranae was assessed as part of the BEEHEAL project during a 2-year study period (2018-2019). Monitoring took place in six apiaries located in four Mediterranean continental apiaries: France, Israel, Portugal and Spain, and two insular apiaries located on Ouessant (France) and on São Miguel (Azores archipelago, Portugal). Besides honeybee sample collection every two months, parameters on colony strength, honey production, beekeeping management and climate were recorded. The pathogen was not detected on São Miguel Island during the study. Infection at the apiary level showed that most of the colonies were positive to N. ceranae except in mainland France, where the percentage of infected colonies was the lowest and it fluctuated, with an increase in August in both years. The percentage of infected honeybees per colony varied among apiaries, with Israel having the highest mean levels (32.39%). Colonies in Spain and Portugal showed a similar pattern with the highest levels of infection in April 2018. Ouessant Island had a high percentage of infected honeybees in 2018 and it decreased on 2019. Temperature was not correlated with N. ceranae infection, although high temperatures combined with high relative humidity were associated to high infection levels in Israel. Intra-colony percentage of infection was positively correlated with honeybee population when all apiaries were analysed together. Honey production was not related with N. ceranae infection. Regarding queen replacement, infection levels in spring 2019 in mainland Portugal and Spain decreased in those colonies that had replaced the queen the previous summer (August 2018). Colony mortality varied amo ng apiaries, with mainland France and Spain showing the highest colony losses (47.6% and 39.1%, respectively). However, this parameter was not correlated with the levels of infection. These results allowed to compare the N. ceranae phenology among different countries showing that the infection is dependent on many factors. This information could assist in the development of beekeeping management strategies.
- Interactions between prevalent pathogens of honeybees (Apis mellifera)Publication . Jabal-Uriel, Clara; Bonjour-Dalmon, Anne; Chejanovsky, Nor; Higes, Mariano; Le Conte, Yves; Lopes, Ana; Pinto, M. Alice; Reyes-Carreño, Maritza; Soroker, Victoria; Martín-Hernández, RaquelThe aim of the BEEHEAL project was to determine the phenology and interactions of the microsporidia Nosema ceranae and 5 highly prevalent and pathogenic honeybee viruses (DWV, BQCV, ABPV, IAPV and CBPV) in four Mediterranean countries: France, Israel, Portugal and Spain. The prevalence and phenology of these pathogens was monitored in four continental apiaries (one in each country) and three insular apiaries located on Ouessant Island (France) and on São Miguel and Santa Maria Islands (Portugal). Colonies were sampled during a 2-year study period, and honeybees were collected and analysed once every two months. Microsporidia detection was performed by conventional triplexPCR whereas viral loads were determined by RT-qPCR of the positive samples. Israel had the highest percentage of honeybees infected by Ǥ ” , followed by Spain, continental Portugal, and Ouessant, whereas the levels were the lowest in France mainland and it was not detected on the Portuguese Islands. The countries with the higher prevalence of viruses were France and Israel whereas the apiaries in Portugal and Spain had a lower percentage of colonies and individual honeybees infected by viruses. Moreover, CBPV was not detected in the latter countries. The viral load of colonies and honeybees coinfected with twoor more viruseswas positively correlated. Data analysedfrom all apiariestogether showed a negative correlation between the percentage of infected N. ceranae honeybees and the viral load. However, there was no correlation at the apiary level between N. ceranae and the viral load when analysed separately. Cross-tabulations showed a correlation in coinfection between N. ceranae with any of the three viruses: BQCV, IAPV and CBPV. In fact, honeybees coinfected by the microsporidia and one of these viruses had a higher viral load than honeybees infected only with the virus. This study showed that pathogen interactions differs greatly among countries and serves as a starting point for the development of epidemiological studies to unravel pathogen coinfections.
- A survey of important honey bee viruses in the Azores archipelagoPublication . Lopes, Ana; Miranda, Joachim; Martín-Hernández, Raquel; Pinto, M. AliceThe Azores archipelago is an interesting place to study honey bee viruses and their associations with Varroa destructor, given that there are islands with (Pico, Flores, and Faial) and without the mite. With the goal of establishing a baseline for future viruses studies, here we sampled 494 colonies distributed across eight islands, in the summers of 2014/2015 and 2020. These samples were screened for the most important honey bee viruses (BQCV, CBPV, SBV, BeeMLV, and the LSV-, DWV-, and AKI-complexes) and the viral loads were determined by RT-qPCR for the positive samples. Of the seven viruses, only BeeMLV and the AKI-complex were not detected. The BQCV was found on all the islands with a high prevalence (>79%) with a wide range of viral loads. São Miguel showed the lowest median BQCV loads (3.18x10͖copies/µL) and Terceira the highest (6.19x10͗copies/µL). In the 2020 sampling,the BQCV loads showed a significant increase on São Jorge and Santa Maria. The LSV was also found on all the islands. Notably, the LSV prevalence increased signiƤcantly in 2020, but not the viral loads. Only Faial and Pico (both with varroa) had SBV, but there was no statistical difference in viral loads between the two islands. CBPV was detected on Pico, São Miguel, Graciosa, Terceira, and Faial, but only in a few colonies (5.3%), although with high viral loads. DWV was never found on São Jorge and Terceira in either sampling period. With the exception of Faial, DWV prevalence was higher on the islands with varroa (Pico and Flores) than on the varroa-free islands. The viral loads were not statistically different between islands with and without varroa for any of the viruses, suggesting that varroa is not the only factor contributing to infection. This study suggests that, in addition to varroa- and varroa-free honey bee populations, some of the islands seem to harbor populations that are also free of some important viruses like DWV, CBPV, SBV, BeeMLV and AKI-complex.
- Large Scale Mitochondrial Evidence of C-lineage Dominance in European Honey BeesPublication . Li, Fernanda; Lopes, Ana; Costa, Maíra; Henriques, Dora; Quaresma, Andreia; Yadró García, Carlos A.; Albo, Alexandre; Čereškienė, Laima Blažytė; Brodschneider, Robert; Brusbardis, Valters; Carreck, Norman L.; Charistos, Leonidas; Chlebo, Robert; Coffey, Mary F.; Dahle, Bjørn; Danneels, Ellen; Dobrescu, Constantin; Dupleix-Marchal, Anna; Filipi, Janja; Gajda, Anna; Gratzer, Kristina; Groeneveld, Linn Fenna; Hatjina, Fani; Johannesen, Jes; Kolasa, Michal; Körmendy-Rácz, János; Kovačić, Marin; Kristiansen, Preben; Martikkala, Maritta; McCormack, Grace P.; Martín-Hernández, Raquel; Pavlov, Borce; Poirot, Benjamin; Pietropaoli, Marco; Radev, Zheko; Raudmets, Aivar; René-Douarre, Vincent; Roessink, Ivo; Škerl, Maja Ivana Smodiš; Soland, Gabriele; Titera, Dalibor; Steen, Jozef van der; Varnava, Andri; Vejsnæs, Flemming; Fedoriak, Mariia M.; Zarochentseva, Oksana; Webster, Matthew T.; Graaf, Dirk C.; Pinto, M. AliceIn Europe, distribution of the several endemic honey bee (Apis mellifera) subspecies has suffered a considerable shift in the last century. In particular, beekeepers tend to favour subspecies of Eastern European ancestry (C-lineage), such as the Italian honey bee (A. m. ligustica), due to their perceived docility and high honey production. As a result, large scale migratory beekeeping and trade of C-lineage queens have exposed the native European honey bees to introgression and replacement, jeopardizing their genetic integrity and locally adapted traits. The maternally-inherited and highly polymorphic mitochondrial intergenic region tRNAleu-cox2 is routinely used for the assessment of honey bee diversity and introgression at large geographical scales. In this study, we conducted a survey on tRNAleu-cox2 variation in more than 1300 colonies from 33 European countries to assess current status of mitochondrial diversity patterns in Europe. Total genomic DNA was extracted from the bee thorax followed by PCR amplification of the tRNAleu-cox2 region and sequencing. The haplotypes were identified using alignments in MEGA 11. A clear dominance of C-lineage haplotypes was found (65%). The most prevalent C-lineage haplotype was C2 (54%) followed by C3 (23%) and C1(22%). Contrary to this trend, in Portugal, Spain and Ireland, less than 10% of the colonies exhibit the C-haplotype. Furthermore, the analysed apiaries in isolated protected areas (n=7) revealed a high proportion of colonies of western European (M-lineage) ancestry (80%) with M4 as the most frequent haplotype. African haplotypes (A lineage) were also found, albeit at lower frequencies, and were mainly concentrated in the Iberian Peninsula (9%). Intensive queen breeding and migratory beekeeping is homogenizing the gene pool of European bee populations. This survey of honey bee maternal diversity across Europe highlights the alarming dominance of C-lineage haplotypes and underscores the importance of conservation apiaries, as they have effectively preserved the autochthonous M-lineage subspecies in different countries. This work was conducted in the framework of the project Better-B, funded by the European Union, the Swiss State Secretariat for Education, Research, and Innovation, and UK Research and Innovation under the UK government's Horizon Europe funding guarantee (grant number 10068544).
- Incense honey (Pittosporum undulatum) in the Azores: Botanical authentication by using real-time PCR approachPublication . Lopes, Ana; Moura, Monica B.M.V.; Grazina, Liliana; Costa, Joana; Amaral, Joana S.; Pinto, M. Alice; Mafra, IsabelHoney is a widely consumed food and much appreciated for its nutritional, organoleptic, and health properties. The honey produced in the Azores archipelago can be categorised as incense honey or multifloral honey. Incense honey classification should account with >30% from Pittosporum undulatum pollen grains and it is generally perceived as a unique and high-quality product and, consequently, susceptible to be adulterated through incorrect labelling or admixing with low-cost and low-quality honeys. Therefore, assessing the authenticity of such highly appreciated honey is a key issue for its valorisation. In this work, a real-time PCR method targeting the ITS region was proposed for the first time to detect P. undulatum species. To that end, the DNA extracted from the leaves of P. undulatum from other endemic species (e.g. Eucaliptus spp., Acacia spp., Trifolium spp., Castanea sativa Mill., Hydrangea macrophylla, Rhododendro indicum, Hedychium gardnerianum, Pericallis malvifolia) of the Azores and mainland Portugal was used to test the cross-reactivity of the ITS primers by qualitative PCR, revealing full specificity for P. undulatum. Posteriorly, a realtime PCR approach was proposed, exhibiting high analytical performance (PCR efficiency= 97.4 ± 6.4%, R͖= 0.991 ± 0.003) and a limit of quantification of 0.01 pg of incense DNA. The method was successfully applied to the 22 honey samples, from which incense was detected in all 9 monofloral incense honeys and in 5 out of 10 multifloral samples from the Azores. Generally, the quantitative results for incense DNA were in good agreement with the melissopalynological data, showing that all samples, except two, were according to their labelled statements. Herein, a new, simple, cost-effective and reliable molecular approach was proposed to authenticate and valorise the Azores honey.
- Bioinformatics pipeline to evaluate patterns of diversity in detoxification genes in Western Honey BeePublication . Barbosa, Daniela; Li, Fernanda; Bashir, Sana; Lopes, Ana; Yadró García, Carlos A.; Quaresma, Andreia; Rufino, José; Rosa-Fontana, Annelise; Verbinnen, Gilles; Graaf, Dirk C.; Smet, Lina de; Pinto, M. Alice; Henriques, DoraThe Western honey bee, Apis mellifera, displays significant genetic diversity in detoxification genes, which is pivotal for environmental adaptation and resilience. Herein, we developed a bioinformatics pipeline to investigate patterns of diversity in these genes, focusing on single nucleotide polymorphisms (SNPs) across A. mellifera populations, with variant annotation performed using both snpEff and the Variant Effect Predictor (VEP). Our pipeline integrates GATK, VCFtools, PLINK, bcftools, snpEff, and VEP to process genomic data systematically. Regions of interest were defined in a BED file for variant filtering. Using GATK, SNPs were extracted from a VCF file and conversion to PLINK format for population genetics analyses. Variants were filtered by minor allele frequency (MAF) and population differentiation (FST index) to identify SNPs with considerable. Variants were annotated with snpEff and VEP to predict functional impacts, enabling a comparative analysis of their annotation consistency and depth. Custom scripts were developed to map SNPs to detoxification genes, quantify SNP density, and integrated gene descriptions and lineage data. The resulting data were visualized using a combination of and generate different graphs using ggplot2 and chromoMap for chromossomal maps. Quality control steps were applied through the pipeline ensuring data reliability. Our findings reveal distinct SNP patterns in detoxification genes, highlighting candidate SNPs associated with A. mellifera subspecies-specific adaptations. The comparison of snpEff and VEP annotations provides insights into their strengths and limitations, which can help optimize software selection for genomic studies. This pipeline offers a reproducible framework for studying genetic diversity in A. mellifera that is adaptable to other species, advancing conservation and evolutionary genomics.