Browsing by Author "Dalmon, Anne"
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- BeeHeal: promoting bee health for sustainable agriculturePublication . Martín-Hernández, Raquel; Lopes, Ana; Chevjanovsky, Nor; Dalmon, Anne; Higes, Mariano; Le Conte, Yves; Pinto, M. Alice; Reyes-Carreño, Maritza; Soroker, VictoriaDuring the last years there is an alarming increase in the collapse of honeybee colonies (Apis mellifera) where bee parasites and pathogens like Varroa destructor mites, the microsporidia Nosema ssp. and viruses have played the leading role. Colony decline might compromise not only food security but also present and future income to the growers. Hence, four institutions from Mediterranean area have joined forces to put toghether a project entitled, “BEEHEAL: Promoting bee health for sustainable agriculture”. BEEHEAL is a collaborative research between Centro de Investigación Apícola y Agroambiental de Marchamalo - CAR (Spain), Centre de recherche Provence-Alpes-Côte d’Azur Unité: Abeilles et Environnement - INRA (France), Agricultural Research Organization, The Volcani Center - ARO (Israel) and Mountain Research Center (CMO), Polytechnic Institute of Bragança (Portugal). The aim of this project is to determine the phenology and interaction of the microsporidia Nosema ceranae and viruses including acute bee paralysis virus (ABPV), Israeli acute paralysis virus (IAPV), Black queen cell virus (BQCV) Chronic bee paralysis virus (CBPV) and Deformed wing virus (DWV), in Spain, France, Portugal and Israel. The findings of this project, which involves an active and unique cooperation among partners representing Mediterranean countries which encompasses a wide range of environmental and beekeeping management conditions, will contribute to ameliorate the damage caused by the expansion of N. ceranae through a rational implementation of existing treatments to avoid emergence of synergistic pathogens that accelerate colony collapse compromising food security. This project started at 2017 and it will end in 2020. BEEHEAL is funded through the ARIMNet2 (2016) Call by the following funding agencies: INIA (Spain), ANR (France), MOARD (Israel), and FCT (Portugal). This presentation will detail the tasks that are ongoing in the BEEHEAL project.
- Beeheal: standardization of laboratory methods for sample processing, nucleic acids extraction and PCR for microsporidia and viruses analysisPublication . Martín-Hernández, Raquel; Benito, M.; Chejanovsky, Nor; Le Conte, Yves; Dalmon, Anne; Higes, Mariano; Pinto, M. Alice; Reyes-Carreño, Maritza; Soroker, VictoriaBEEHEAL is a project designed to determine the phenology and interaction of Nosema ceranae and viruses in four Mediterranean countries: Spain, France, Portugal and Israel, including some territories where Varroa destructor is not present (Azores and Ouessant islands). This will allow us to study and compare the interactions between pathogens in a wide range of hosts, beekeeping and climatic conditions. The honey bee samples collected along the year in the different countries will be analysed for pathogens in three laboratories. This requires a standardization of methods to compare the results in order to assign the effect of every variable in a reliable way. To that end, the participating laboratories have been working together to establish the sampling methodology, the conservation of the samples, the nucleic acids extraction and the PCR analysis. We analyzed the sample processing for nucleic acid extraction on TE buffer (with or without Proteinase K), CTAB buffer or commercial kits (Qiagen). The maceration of bees (either individually or in composite samples) in TE buffer and posterior incubation at 96ºC for 20 minutes showed a good sensibility level and good value for N. ceranae DNA extraction. This method also allowed the conservation of RNA at -80ºC for a month in the TE solution for later RNA extraction. A joint protocol for sample processing, DNA and RNA extraction and PCR analysis has been developed but adjusted to the particular conditions and equipment of each laboratory. The standardization of methods to be implemented by each participating laboratory will avoid the biases on conclusions based on the diverse methods applied.
- Mitochondrial and nuclear diversity of colonies of varying origins: contrasting patterns inferred from the intergenic tRNAleu-cox2 region and immune SNPsPublication . Henriques, Dora; Lopes, Ana; Chejanovsky, Nor; Dalmon, Anne; Higes, Mariano; Jabal-Uriel, Clara; Le Conte, Yves; Reyes-Carreño, Maritza; Soroker, Victoria; Martín-Hernández, Raquel; Pinto, M. AliceIn this study, we gathered sequence data from the tRNAleu-cox2 intergenic mitochondrial (mtDNA) region concurrently with single nucleotide polymorphism (SNP) data from 91 loci of nuclear DNA (ncDNA). The data was obtained from 156 colonies sampled in six apiaries from four countries. The full dataset was analysed and discussed for genetic patterns with a focus on cytonuclear diversity and admixture levels.
- Nosema ceranae in Apis mellifera: a 12 years postdetection perspectivePublication . Martín-Hernández, Raquel; Bartolomé, Carolina; Chejanovsky, Nor; Le Conte, Yves; Dalmon, Anne; Dussaubat, Claudia; García-Palencia, Pilar; Meana, Aranzazu; Pinto, M. Alice; Soroker, Victoria; Higes, MarianoNosema ceranae is a hot topic in honey bee health as reflected by numerous papers published every year. This review presents an update of the knowledge generated in the last 12 years in the field of N. ceranae research, addressing the routes of transmission, population structure and genetic diversity. This includes description of how the infection modifies the honey bee’s metabolism, the immune response and other vital functions. The effects on individual honey bees will have a direct impact on the colony by leading to losses in the adult’s population. The absence of clear clinical signs could keep the infection unnoticed by the beekeeper for long periods. The influence of the environmental conditions, beekeeping practices, bee genetics and the interaction with pesticides and other pathogens will have a direct influence on the prognosis of the disease. This review is approached from the point of view of the Mediterranean countries where the professional beekeeping has a high representation and where this pathogen is reported as an important threat.
- Projeto BEEHEAL: promover a saúde da abelha para uma agricultura sustentávelPublication . Lopes, Ana; Pinto, M. Alice; Chevjanovsky, Nor; Soroker, Victoria; Le Conte, Yves; Dalmon, Anne; Reyes-Carreño, Maritza; Higes, Mariano; Martín-Hernández, RaquelO BEEHEAL, com o título original “Promoting bee health for sustainable agriculture”, é um projeto internacional colaborativo aprovado no âmbito da Ação ERA-Net ARIMNet2 (Coordination of Agricultural Research in the Mediterranean). O projeto é coordenado por Raquel Martín- Hernández, investigadora do “Centro de Investigación Apícola y Agroambiental de Marchamalo” (CAR), Espanha. Para além deste centro de investigação, representado por Raquel Martín-Hernández e Mariano Higes, o consórcio inclui mais três instituições, nomeadamente: o Centro de Investigação de Montanha (CIMO) do Instituto Politécnico de Bragança, representado por M. Alice Pinto e Ana Rita Lopes, o “Centre de Recherche Provence-Alpes-Côte d’Azur Unité: Abeilles et Environnement do “Institut National de la Recherche Agronomique” (INRA), França, representado por Yves Le conte, Anne Dalmon e Maritza Maritza Reyes-Carreno, e o “Volcani Center” da “Agricultural Research Organization” (ARO), Israel, representado por Nor Chevjanovsky e Victoria Soroker. As populações de abelha melífera (Apis mellifera L.) têm vindo a sofrer perdas acentuadas em todo o mundo. Estas perdas estão relacionado com vários factores, que podem atuar sozinhos ou em combinação, incluindo (i) propagação de parasitas e agentes patogénicos exóticos , como por exemplo o ácaro ectoparasita Varroa destructor, o qual serve de vetor de transmissão de vários vírus, e o fungo microsporídeo Nosema ceranae, (ii) exposição das colónias a agro-químicos, (iii) má nutrição, (iv) alterações climáticas, entre outros (vanEngelsdorp & Meixner, 2010; Potts et al., 2010).
- A SNP assay for assessing diversity in immune genes in the honey bee (Apis mellifera L.)Publication . Henriques, Dora; Lopes, Ana; Chejanovsky, Nor; Dalmon, Anne; Higes, Mariano; Jabal-Uriel, Clara; Le Conte, Yves; Reyes-Carreño, Maritza; Soroker, Victoria; Martín-Hernández, Raquel; Pinto, M. AliceWith a growing number of parasites and pathogens experiencing large-scale range expansions, monitoring diversity in immune genes of host populations has never been so important because it can inform on the adaptive potential to resist the invaders. Population surveys of immune genes are becoming common in many organisms, yet they are missing in the honey bee (Apis mellifera L.), a key managed pollinator species that has been severely affected by biological invasions. To fill the gap, here we identified single nucleotide polymorphisms (SNPs) in a wide range of honey bee immune genes and developed a medium-density assay targeting a subset of these genes. Using a discovery panel of 123 whole-genomes, representing seven A. mellifera subspecies and three evolutionary lineages, 180 immune genes were scanned for SNPs in exons, introns (< 4 bp from exons), 3’ and 5´UTR, and < 1 kb upstream of the transcription start site. After application of multiple filtering criteria and validation, the final medium-density assay combines 91 quality-proved functional SNPs marking 89 innate immune genes and these can be readily typed using the high-sample-throughput iPLEX MassARRAY system. This medium-density-SNP assay was applied to 156 samples from four countries and the admixture analysis clustered the samples according to their lineage and subspecies, suggesting that honey bee ancestry can be delineated from functional variation. In addition to allowing analysis of immunogenetic variation, this newly-developed SNP assay can be used for inferring genetic structure and admixture in the honey bee.
- A SNP assay for assessing diversity in immune genes in the honey bee (Apis mellifera L.)Publication . Henriques, Dora; Lopes, Ana; Chejanovsky, Nor; Dalmon, Anne; Higes, Mariano; Jabal-Uriel, Clara; Le Conte, Yves; Reyes-Carreño, Maritza; Soroker, Victoria; Martín-Hernández, Raquel; Pinto, M. AliceWith a growing number of parasites and pathogens experiencing large-scale range expansions, monitoring diversity in immune genes of host populations has never been so important because it can inform on the adaptive potential to resist the invaders. Population surveys of immune genes are becoming common in many organisms, yet they are missing in the honey bee (Apis mellifera L.), a key-stone species that has been severely affected by biological invasions. To fill the gap, here we identified single nucleotide polymorphisms (SNPs) in honey bee immune genes and then developed an assay to be genotyped using the high-sample-throughput iPLEX MassARRAY system that can be readily used for population surveys. The assay was constructed using SNPs detected in whole-genome scans of 123 individuals originating from a wide geographical area, representing seven A. mellifera subspecies and three evolutionary lineages (M-Western European, C- Eastern European, A- African). In this dataset, a total of 35,782 SNPs distributed through 180 genes were found. Only polymorphic SNPs (MAF>0.05) SNPs located in putatively functional regions were retained. Other filtering criteria inked to the MassARRAY® MALDI-TOF genotyping system were used and an assay with 107 SNPs was obtained. A total of 16 SNPs were discarded either due to inconsistent calls and/or misidentification of heterozygous positions. The final assay contains 91 quality-proved functional SNPs covering 89 innate immune genes. The 89 genes belong to several families and pathways, such as IMD, JAK-STAT, Toll and RNAi. This gene set also includes genes that have been found to be expressed in honey bees infected with the viruses SINV, IAPV, DWV and Nosema spp. This medium-density-SNP assay was applied to 156 samples from four countries (Portugal, Spain, France and Israel) and the admixture analysis clustered the samples according to their lineage and subspecies, suggesting that the immune SNPs can be also used to reconstruct population structure. This newly-developed SNP assay can be applied to monitoring diversity in immune genes, identifying the genetic basis of disease susceptibility, and even inferring genetic structure.
- Virus prevalence in egg samples collected from naturally selected and traditionally managed honey bee colonies across europePublication . Bouuaert, David Claeys; De Smet, Lina; Brunain, Marleen; Dahle, Bjørn; Blacquiere, Tjeerd; Dalmon, Anne; Dezmirean, Daniel; Elen, Dylan; Filipi, Janja; Giurgiu, Alexandru; Gregorc, Aleš; Kefuss, John; Locke, Barbara; Miranda, Joachim; Oddie, Melissa; Panziera, Delphine; Parejo, Melanie; Pinto, Maria Alice; Graaf, Dirk C.Monitoring virus infections can be an important selection tool in honey bee breeding. A recent study pointed towards an association between the virus-free status of eggs and an increased virus resistance to deformed wing virus (DWV) at the colony level. In this study, eggs from both naturally surviving and traditionally managed colonies from across Europe were screened for the prevalence of different viruses. Screenings were performed using the phenotyping protocol of the ‘suppressed in ovo virus infection’ trait but with qPCR instead of end-point PCR and a primer set that covers all DWV genotypes. Of the 213 screened samples, 109 were infected with DWV, 54 were infected with black queen cell virus (BQCV), 3 were infected with the sacbrood virus, and 2 were infected with the acute bee paralyses virus. It was demonstrated that incidences of the vertical transmission of DWV were more frequent in naturally surviving than in traditionally managed colonies, although the virus loads in the eggs remained the same. When comparing virus infections with queen age, older queens showed significantly lower infection loads of DWV in both traditionally managed and naturally surviving colonies, as well as reduced DWV infection frequencies in traditionally managed colonies. We determined that the detection frequencies of DWV and BQCV in honey bee eggs were lower in samples obtained in the spring than in those collected in the summer, indicating that vertical transmission may be lower in spring. Together, these patterns in vertical transmission show that honey bee queens have the potential to reduce the degree of vertical transmission over time.