Preservation of Honey Bee (Apis mellifera L.) Semen

L’abeille domestique (Apis mellifera Linnaeus) joue un rôle crucial comme pollinisateur dans l’industrie de l’agriculture. Cependant, durant les dernières décennies, une mortalité des colonies d’abeilles a été observée partout à travers le monde. La conservation du sperme d’abeille est un outil efficace pour sauvegarder la diversité génétique. Sa conservation est possible à température pièce, mais la cryoconservation serait une meilleure méthode pour la conservation à long terme. Notre objectif général est de développer une méthode de cryoconservation de la semence d’abeille. L’hypothèse no.1 était que la cryoconservation de la semence d’abeille est plus efficace à long terme que les températures au-dessus de 0 °C. Nous avons évalué l’efficacité, basé sur la viabilité des spermatozoïdes, de deux températures de conservation: -196 °C et 16 °C. Après un an de conservation, la semence congelée avait une meilleure viabilité comparée à 16°C (76% ± 5% vs 0%; p < 0,05). Par la suite, la spermathèque des reines inséminées avec la semence cryoconservée a été évaluée par la migration des spermatozoïdes ainsi que la viabilité des spermatozoïdes. Il y avait beaucoup de variabilités dans nos résultats. Nous n’avons pas été en mesure de vérifier si l’ajout de la centrifugation après la conservation améliore la fertilité des reines après insémination. Toutefois, nos résultats confirment que la cryoconservation est une technique efficace pour conserver la semence d’abeille à long terme.

Enhancing legume ecosystem services through an understanding of plant–pollinator interplay

Legumes are bee-pollinated, but to a different extent. The importance of the plant– pollinator interplay (PPI), in flowering crops such as legumes lies in a combination of the importance of pollination for the production service and breeding strategies, plus the increasing urgency in mitigating the decline of pollinators through the development and implementation of conservation measures. To realize the full potential of the PPI, a multidisciplinary approach is required. This article assembles an international team of genebank managers, geneticists, plant breeders, experts on environmental governance and agro-ecology, and comprises several sections. The contributions in these sections outline both the state of the art of knowledge in the field and the novel aspects under development, and encompass a range of reviews, opinions and perspectives. The first three sections explore the role of PPI in legume breeding strategies. PPI based approaches to crop improvement can make it possible to adapt and re-design breeding strategies to meet both goals of: (1) optimal productivity, based on an efficient use of pollinators, and (2) biodiversity conservation. The next section deals with entomological aspects and focuses on the protection of the “pest control service” and pollinators in legume crops. The final section addresses general approaches to encourage the synergybetweenfoodproductionandpollinationservicesatfarmerfieldlevel.Twobasic approaches are proposed: (a) Farming with Alternative Pollinators and (b) Crop Design System.

Study of the allergenic profile of pollen from Platanus hybrida in the city of Evora, Portugal

Background and Aim: Although grasses and olive are the most relevant allergenic species in the Alentejo region, aggravation of allergic symptoms in the early spring, unrelated with those species pollen seasons, has been reported, particularly in urban environment. Plane trees, hence pollen, are highly abundant in the city of Évora, nonetheless allergen pollen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Platanus hybrida, one of the most representative species in Evora showing pollination prior to the main pollen season in Alentejo. Methods: Pollen from Platanus hybrida and Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Sensitization and cross-reactivity was assessed by solid phase immunoblot. Results: Half of the patient exhibited sensitization to pollen extracts of P. Hybrida. Western blot have shown several immunoreactive bands in the Mr 10-90 kDa range. Immunoreactive bands were also observed in the protein profile according to the pI in the pI range 4.0-6.1. Cross-reactivity of P. hybrida with D. glomerata was found. Although several bands are common to D. glomerata, a band with ~50kDa was observed in P. hybrida but not in D. glometata. Conclusion: These results evidenced allergens found in P. hybrida pollen. Moreover, cross–reactivity between P. hybrida and highly allergenic species such as D. glomerata was found which probably contributes for aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by FEDER through the “Programa Operacional Fatores de Competitividade – COMPETE” (Strategic projects of ICAAM and ICT 2013-2015).

Allergenic profile of Quercus rotundifolia pollen in Alentejo, Portugal

Background and Aim: Grasses and olive are the most relevant allergenic species in the Alentejo region. However, aggravation of allergic symptoms has been reported in the early spring, before grass and olive pollen seasons. Quercus pollen is the most abundant pollen type in the early spring in Alentejo, nonetheless its allergen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Quercus rotundifolia among the most representative species showing pollination in April, prior to the main pollen season in Alentejo. Methods: Pollen from Quercus rotundifolia, Olea europaea and Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Extract from Quercus ilex pollen was kindly offered by Bial. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Sensitization and cross-reactivity was assessed by solid phase immunoblot. Results: Most of the patient evidenced sensitization to pollen extracts of Q. rotundifolia. Protein profile of Q. rotundifolia has shown several bands in the Mr 10-90 kDa, mostly overlapping with Q. ilex. Western blot have shown several immunoreactive bands. Immunoreactive bands were also observed in the protein profile according to the pI in the range 4.0-6.1. Cross-reactivity between Q. rotundifolia with O. europaea and D. glomerata was found. Conclusion: These results evidenced allergens found in Q. rotundifolia pollen. It also shows that protein profile of Q. rotundifolia and Q. ilex are mostly alike suggesting that similarities in allergen profile are expected. Moreover, cross–reactivity between Q. rotundifolia and highly allergenic species such as O. europaea and D. glomerata was found which probably contributes to the aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by FEDER through the “Programa Operacional Fatores de Competitividade – COMPETE” (Strategic projects of ICAAM and ICT 2013-2015). We also aknowledge Bial-Aristegui for supplying pollen and extract samples of Q. ilex.

Perfil em alergénios do pólen de Platanus hybrida

Objetivos: O plátano (Platanus hybrida) é uma árvore frequentemente utilizada em ambiente urbano, com fins ornamentais. Sendo uma árvore de grande porte, produz pólen em grande quantidade. Embora seja responsável por níveis de exposição a pólen elevados no início da primavera, que são coincidentes com queixas da população, o seu potencial alergénico está pouco caracterizado. Este trabalho teve, assim, como objetivo caracterizar o perfil em alergénios do pólen de plátano na cidade de Évora, Alentejo. Métodos: Prepararam-se extratos de amostras de pólen de Platanus hybrida ou Dactylis glomerata utilizando tampão bicarbonato. Os extratos foram liofilizados e conservados a -80ºC. O conteúdo em proteínas foi determinado pelo método de Bradford. O perfil em alergénios foi avaliado por western blot utilizando soros humanos (obtidos mediante consentimento informado de doentes do Hospital do Espírito Santo de Évora – HESE). Resultados: Observou-se teste positivo a P. hybrida em metade dos soros testados. O perfil em proteínas de P. hybrida exibiu diversas bandas imunorreativas com massas moleculares compreendidas entre 10-90 kDa e com pI no intervalo 4,4-7,0. Foram encontradas imunorreativas comuns a Q. rotundifólia e/ou a D. glomerata. Duas bandas identificadas na gama de 50kDa e 60 kDa parecem específicas de P. hybrida. Também se registou reatividade cruzada com D. glomerata. Conclusões: Este trabalho evidencia alguns alergénios encontrados em pólen de P. hybrida. Para além disso mostra ainda a existência de reatividade cruzada com pólen de gramíneas. Estes resultados sugerem que o pólen de plátano, dada a sua grande abundância na cidade de Évora, poderá contribuir para o agravamento a sintomatologia da população que sofre de polinose, em particular no início da primavera. Agradecimentos: Este trabalho foi financiado por fundos do FEDER através do Programa Operacional Fatores de Competitividade – COMPETE”. Um agradecimento especial ao nosso colega, já falecido, Prof. Rui Brandão, pelo estímulo que deu a este trabalho e pela sua dedicação para a implementação e desenvolvimento da Aerobiologia na Universidade de Évora. Temos a honra de dedicar este trabalho à sua memória. Background and Aim: Although grasses and olive are the most relevant allergenic species in the Alentejo region, aggravation of allergic symptoms in the early spring, unrelated with those species pollen seasons, has been reported, particularly in urban environment. Plane trees, hence pollen, are highly abundant in the city of Évora, nonetheless allergen pollen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Platanus hybrida, one of the most representative species in Evora showing pollination prior to the main pollen season in Alentejo. Methods: Pollen from Platanus hybrida, Quercus rotundifolia or Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Results: Protein profile of P. Hybrida has shown several bands in the Mr 10-90 kDa. Western blot have shown several immunoreactive bands. Protein profile according to the pI showed immunoreactive bands in the pI range 4.0-6.1. Cross-reactivity of P. hybrida with Q. rotundifolia and D. glomerata was found. Conclusion: These results evidenced allergens found in P. hybrida pollen. Moreover, cross–reactivity between P. hybrida and highly allergenic species such as D. glomerata was found which probably contributes for aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by “FEDER - Programa Operacional Factores de Competitividade – COMPETE”. A special acknowledgment to our colleague Prof. Rui Brandão, deceased, for his dedication to the present work, to the implantation and development of Aerobiology in the University of Évora. We have the honour of dedicating this work to the memory of Prof. Rui Brandão.

Perfil em alergénios do pólen de Quercus rotundifólia

Background and Aim: Grasses and olive are the most relevant allergenic species in the Alentejo region. However, aggravation of allergic symptoms has been reported in the early spring, before grass and olive pollen seasons. Quercus pollen is the most abundant pollen type in the early spring in Alentejo, nonetheless its allergen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Quercus rotundifolia the most representative species showing pollination in April, prior to the main pollen season in Alentejo. Methods: Pollen from Quercus rotundifolia, Olea europaea and Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Extract from Quercus ilex pollen was kindly offered by Bial. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Results: Protein profile of Q. rotundifolia has shown several bands in the Mr 10-90 kDa, mostly overlapping with Q. ilex. Western blot have shown several immunoreactive bands. Protein profile according to the pI showed immunoreactive bands in the pI range 4.0-6.1. Cross-reactivity of Q. rotundifolia with O. europaea and D. glomerata was found. Conclusion: These results evidenced allergens found in Q. rotundifolia pollen. It also shows that protein profile of Q. rotundifolia and Q. ilex are mostly alike suggesting that similarities in allergen profile are expected. Moreover, cross–reactivity between Q. rotundifolia and highly allergenic species such as O. europaea and D. glomerata was found which probably contributes for aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by “FEDER - Programa Operacional Factores de Competitividade – COMPETE”. A special acknowledgment to our colleague Prof. Rui Brandão, deceased, for his dedication to the present work, to the implantation and development of Aerobiology in the University of Évora. We have the honour of dedicating this work to the memory of Prof. Rui Brandão.