Biosecurity measures in European beekeeping.

Emerging pathogens of honey bees represent an important threat to the development of the beekeeping sector. The implementation of biosecurity measures in beekeeping (BMBs) plays an essential role in supporting honey bee health within the beekeeping sector. A group of experts, in collaboration with the BPRACTICES (Grant Agreement No. 696231, European Research Area on Sustainable Animal Production Systems [ERA-Net SusAn]) project partners, has provided the definition of BMBs. Thus, BMBs are all those operational activities implemented to control the risk of introduction and spread of specific honey bee disease agents. In this paper, the BMBs in the European beekeeping context are identified for the most relevant honey bee diseases in Europe: varroosis, American foulbrood (AFB), European foulbrood (EFB), nosemosis and aethinosis. Moreover, BMBs were classified in â€Ëœcategories' adapted to consider productivity and the â€ËœOne Health' approach: human health, honey bee health and protection of the environment. The 84 BMBs described by the panel of experts were ranked according to the priority score attributed. The implementation of BMBs represents an essential step forwards to increase the resilience and sustainability of European beekeeping.

L'émergence de nouveaux agents pathogènes chez les abeilles mellifères représente une menace importante pour le développement du secteur apicole. La mise en oeuvre de mesures de biosécurité en apiculture est déterminante pour préserver la santé des abeilles mellifères dans les élevages. En collaboration avec des partenaires du projet BPRACTICES (convention de subvention n° 696231, programme ERA-Net SusAn [European Research Area on Sustainable Animal Production Systems]), un groupe d'experts a élaboré un cadre définissant ces mesures de biosécurité. Sont définies comme mesures de biosécurité en apiculture toutes les activités opérationnelles mises en oeuvre pour contrôler le risque d'introduction et de propagation d'agents pathogènes affectant spécifiquement les abeilles mellifères. Les auteurs décrivent les mesures de biosécurité applicables, dans le contexte apicole européen, aux maladies les plus importantes en Europe : la varroose, la loque américaine, la loque européenne, la nosémose et l'infestation par Aethina tumida. En outre, les mesures de biosécurité en apiculture ont été regroupées en « catégories ¼ afin de prendre en compte la productivité et l'approche « Une seule santé ¼ : santé humaine, santé des abeilles mellifères et protection de l'environnement. Les 84 mesures de biosécurité en apiculture décrites par le groupe d'experts ont été classées en fonction du niveau de priorité qui leur a été attribué. La mise en oeuvre de ces mesures représente une étape cruciale pour accroître la résilience et la durabilité de l'apiculture européenne.

Los agentes patógenos emergentes que afectan a la abeja melífera suponen una importante amenaza para el desarrollo del sector apícola. La aplicación de medidas de seguridad biológica dentro de este sector cumple una función esencial para proteger la salud de las abejas. En colaboración con asociados en el proyecto BPRACTICES (acuerdo de subvención nº 696231, programa ERA-Net SusAn [Espacio Europeo de Investigación - «Sistemas sostenibles de producción animal¼]), un grupo de expertos definió las «medidas de seguridad biológica en apicultura¼ como todas aquellas acciones realizadas para controlar el riesgo de penetración y propagación de agentes patógenos de la abeja melífera. Los autores, situándose en el contexto de la apicultura europea, exponen las medidas de seguridad biológica que ayudan a controlar las principales enfermedades de la abeja melífera en Europa: varroosis, loque americana, loque europea, nosemosis y aethinosis (infestación por el escarabajo de las colmenas). Por otra parte, estas medidas fueron divididas en diferentes «categorías¼ para poder tener en cuenta las cuestiones de productividad y el enfoque de «Una sola salud¼: salud humana, salud de la abeja melífera y protección del medio ambiente. Las 84 medidas de seguridad biológica en apicultura que describió el cuadro de expertos fueron jerarquizadas en función de una puntuación atribuida por su nivel de prioridad. La aplicación de este tipo de medidas representa un crucial paso adelante para conferir más resiliencia a la apicultura europea y hacerla más sostenible.

Good farming practices in apiculture.

Modern European beekeeping is facing numerous challenges due to a variety of factors, mainly related to globalisation, agrochemical pollution and environmental changes. In addition to this, new pathogens threaten the health of European honey bees. In that context, correct colony management should encompass a wider vision, where productivity aspects are linked to a One Health approach in order to protect honey bees, humans and the environment. This paper describes a novel tool to be applied in beekeeping operations: good beekeeping practices (GBPs). The authors ranked a list of GBPs scored against their importance and validated by an international team, including researchers, national animal health authorities and international beekeepers' associations. These activities were carried out in the project 'BPRACTICES', approved within the transnational call of the European Research Area Network on Sustainable Animal Production (ERA-NET SusAn) in the Horizon 2020 Research and Innovation Programme of the European Union. This study, created through an international collaboration, aims to present an innovative and implementable approach, similar to applications already adopted in other livestock production systems.

L'apiculture moderne européenne est confrontée à de nombreuses difficultés dues à divers facteurs, pour la plupart liés à la mondialisation, à la pollution agrochimique et à la modification de l'environnement. À ces facteurs s'ajoute l'émergence de nouveaux agents pathogènes qui menacent la santé des abeilles mellifères d'Europe. Dans ce contexte, une gestion appropriée des colonies d'abeilles devrait reposer sur une vision plus large, dans laquelle les aspects relevant de la productivité sont examinés suivant une approche « Une seule santé ¼ afin de protéger les abeilles mellifères, les humains et l'environnement. Les auteurs décrivent un nouvel outil destiné à l'apiculture : les bonnes pratiques apicoles. Ils ont évalué et classé par ordre d'importance une liste de bonnes pratiques apicoles validées par une équipe internationale composée de chercheurs, d'autorités nationales de la santé animale et d'associations internationales d'apiculteurs. Ces activités ont été conduites dans le cadre du projet « BPRACTICES ¼, proposition retenue suite à l'appel à projets transnationaux du réseau ERA­NET SusAn (European Research Area Network on Sustainable Animal Production) au sein du Programme Horizon 2020 de l'Union européenne pour la recherche et l'innovation. Conçue sous forme de collaboration internationale, cette étude vise à proposer une approche innovante et pratique, similaire aux applications précédemment adoptées dans d'autres systèmes de production animale.

La apicultura europea hace frente a numerosos problemas resultantes de diversos factores, relacionados principalmente con la mundialización, la contaminación agroquímica y los cambios ambientales, a todo lo cual se suman nuevos patógenos que amenazan la salud de las abejas melíferas europeas. En este contexto, una correcta gestión de las colonias debe traer aparejada una visión más global, en la que las cuestiones de productividad se consideren en clave de «Una sola salud¼ con objeto de proteger tanto a las abejas melíferas como a las personas y el medio ambiente. En este artículo se describe una novedosa herramienta aplicable a la actividad apícola: las buenas prácticas de apicultura. Los autores jerarquizaron una serie de buenas prácticas de apicultura seleccionadas, validadas y puntuadas según su importancia por un equipo internacional que incluía a investigadores, autoridades nacionales de sanidad animal y asociaciones internacionales de apicultores. Este trabajo formaba parte del proyecto «BPRACTICES¼, aprobado con ocasión de la convocatoria internacional abierta por la Red del espacio europeo de investigación en sanidad animal sostenible (ERA­NET SusAn), inscrita a su vez en Horizonte 2020, el programa de investigación e innovación de la Unión Europea. El estudio aquí descrito, fruto de la colaboración internacional, tiene por objeto presentar un planteamiento novedoso y viable, parecido a las aplicaciones ya implantadas en otros sistemas de producción animal.

Diagnosis of American foulbrood in honey bees: a synthesis and proposed analytical protocols.

Worldwide, American foulbrood (AFB) is the most devastating bacterial disease of the honey bee (Apis mellifera). Because the distinction between AFB and powdery scale disease is no longer considered valid, the pathogenic agent has recently been reclassified as one species Paenibacillus larvae, eliminating the subspecies designations Paenibacillus larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens. The creamy or dark brown, glue-like larval remains of infected larvae continue to provide the most obvious clinical symptom of AFB, although it is not conclusive. Several sensitive and selective culture media are available for isolation of this spore-forming bacterium, with the type of samples that may be utilized for detection of the organism being further expanded. PCR methods for identification and genotyping of the pathogen have now been extensively developed. Nevertheless, biochemical profiling, bacteriophage sensitivity, immunotechniques and microscopy of suspect bacterial strains are entirely adequate for routine identification purposes.

Histochemical characterization of cell death in honeybee larvae midgut after treatment with Paenibacillus larvae, Amitraz and Oxytetracycline.

A number of techniques were employed to assess cell death induced in honeybee larvae midgut after per os inoculation of bacterium Paenibacillus larvae var. larvae, the causative agent of American foulbrood disease, and separately with acaricide Amitraz and antibiotic Oxytetracycline. In honeybee larvae exposed to Amitraz, which demonstrates both necrosis and apoptosis, cell death was found in 82% of midgut columnar and in 50% of regenerative epithelial cells, 24 h after treatment. Cell death reduced to 36% in the epithelial cells, 48 h after treatment. In Oxytetracycline-treated larvae, cell death was identified in 40% of midgut epithelial cells, 24 h after inoculation and increased to 55% over the next 24 h. In Paenibacillus -infected larvae, all midgut epithelial cells died. Using ApopTag (Oncor) to label the multiple DNA ends generated by DNA fragmentation showed programmed cell death in 49% of columnar midgut cells 24 h after Amitraz application. Cell death was reduced to 9% over the next 24 h. Our data indicate that cell death could be identified and quantified in situ, using TUNEL techniques. This study also shows that the acaricide Amitraz is a trigger for programmed cell death in the midgut epithelial cells of honeybee larvae, unlike Paenibacillus which induces necrosis only. The data show that immunohistochemical methods are useful for studying in situ tissue pathology, and indicate possibilities for monitoring the effects of infective and chemical environmental stressors on cell death in honeybee larvae tissue.

In situ localization of heat-shock and histone proteins in honey-bee (Apis mellifera l.) larvae infected with Paenibacillus larvae.

The immunohistochemical localization of the heat shock proteins (Hsp70 and Hsp90) and histone protein in healthy and Paenibacillus larvae infected honeybee (Apis mellifera L.) larvae has been studied. Hsp70 was found in the nuclei and the cytoplasm of infected midgut, salivary gland cells and haemocytes, but not in uninfected larvae. Hsp90 was localized in both infected and uninfected cells. Exposed histone proteins were localized in the nuclei of dying uninfected cells undergoing programmed cell death. The distribution of histone protein in uninfected cells of midgut, salivary gland, and other tissues was nuclear and indicative of normal programmed cell death at levels between 1 and 5%. After applying histone protein antibodies to P. larvae infected honeybee larvae, the DAB based reaction product was located in the nuclei or immediate surroundings of all larval cells. The Hsp70, Hsp90 and histone protein distribution patterns are discussed in relation to the morphological, cytochemical and immunocytochemical characteristics of programmed cell death and pathological necrosis. Results produced by methyl green-pyronin staining confirm an elevation of RNA levels in normal programmed cell death and a reduced staining for RNA in necrotic infected cells.

Histopathological and histochemical changes in honeybee larvae (Apis mellifera L.) after infection with Bacillus larvae, the causative agent of American foulbrood disease.

Morphological, histochemical and cytochemical changes were examined in honeybee larvae after infection with the bacterium Bacillus larvae. The results indicate cell necrosis in the midgut epithelium accompanied by increasing cell vacuolization and nuclear pyknosis following per os inoculation with B. larvae. Many autolysosomes were positive for acid phosphatase. Non-vacuolar acid phosphatase activity was also found in lysed cell compartments. No such activity was found in regenerative epithelial cells. Degradation of haemocytes, salivary glands and other tissues was also observed. Histochemical analyses after per cutaneous inoculation with B. larvae of three- and five-day-old honeybee larvae show intense non-vacuolar acid phosphatase activity followed by disintegration of infected salivary glands, epithelial cell cytoplasm and haemocytes.

Programmed cell death in the honey-bee (Apis mellifera L.) larvae midgut.

The histochemical and cytochemical localization of acid phosphatase has been used in an attempt to map the sites of cellular lysis and death. Reaction product was found both in the brush border of the midgut epithelium and in the basal membrane. Vacuolar acid phosphatase activity was found in the regenerative epithelial cells. Extra-cisternal reaction product was associated with the endoplasmic reticulum which was dilated in lysed areas of the cytoplasm. Free acid and alkaline phosphatase activity was found in the basal area of the midgut epithelial cells and the former also occurred in the haemocoel. In the tracheoblastic cells only vacuolar acid phosphatase activity was seen. Chromatin aggregates were distributed throughout the nucleus and the nuclear envelope showed some infolding. Certain mature epithelial cells proved positive for anti-histone associated DNA fragmentation indicative of programmed cell death.