Bee viruses: routes of infection in Hymenoptera

Numerous studies have recently reported on the discovery of bee viruses in different arthropod species and their possible transmission routes, vastly increasing our understanding of these viruses and their distribution. Here, we review the current literature on the recent advances in understanding the transmission of viruses, both on the presence of bee viruses in Apis and non-Apis bee species and on the discovery of previously unknown bee viruses. The natural transmission of bee viruses will be discussed among different bee species and other insects. Finally, the research potential of in vivo (host organisms) and in vitro (cell lines) serial passages of bee viruses is discussed, from the perspective of the host-virus landscape changes and potential transmission routes for emerging bee virus infections

Cold case: The disappearance of Egypt bee virus, a fourth distinct master strain of deformed wing virus linked to honeybee mortality in 1970’s Egypt

In 1977, a sample of diseased adult honeybees (Apis mellifera) from Egypt was found to contain large amounts of a previously unknown virus, Egypt bee virus, which was subsequently shown to be serologically related to deformed wing virus (DWV). By sequencing the original isolate, we demonstrate that Egypt bee virus is in fact a fourth unique, major variant of DWV (DWV-D): more closely related to DWV-C than to either DWV-A or DWV-B. DWV-A and DWV-B are the most common DWV variants worldwide due to their close relationship and transmission by Varroa destructor. However, we could not find any trace of DWV-D in several hundred RNA sequencing libraries from a worldwide selection of honeybee, varroa and bumblebee samples. This means that DWV-D has either become extinct, been replaced by other DWV variants better adapted to varroa-mediated transmission, or persists only in a narrow geographic or host range, isolated from common bee and beekeeping trade routes

A SNP assay for assessing diversity in immune genes in the honey bee (Apis mellifera L.)

With 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.We are deeply indebted to Frank Aguiar, Luís Silva, Edgardo Melo, João Martins, João Melo, Manuel Moura, Manuel Viveiros, and Ricardo Sousa from "Direção Regional da Agricultura e Desenvolvimento Rural dos Açores" (Portugal), and to Laura Garreau, Laurent Maugis, Pascale Sauvage and Jacques Kermagoret, from “Association Conservatoire de l’Abeille Noir Bretonne” (France), for sampling the apiaries in São Miguel, Santa Maria, and Ouessant islands. Genotyping was outsourced to the Epigenetics and Genotyping laboratory, Central Unit for Research in Medicine (UCIM), University of Valencia, Spain. Data analyses were performed using computational resources at the Research Centre in Digitalization and Intelligent Robotics (CeDRI), Instituto Politécnico de Bragança. Ana Rita Lopes is supported by a PhD scholarship (SFRH/BD/143627/2019) from the Foundation for Science and Technology (FCT), Portugal. FCT provided financial support by national funds (FCT/MCTES) to CIMO (UIDB/00690/2020).This research was funded through the projects BEEHAPPY (POCI-01-0145- FEDER-029871, FCT and COMPETE/QREN/EU) and BEEHEAL. BEEHEAL was funded by the ARIMNet2 2016 Call by the following agencies: INIA (Spain), MOARD (Israel), ANR (France) and FCT (Portugal). ARIMNet2 (ERA-NET) received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 618127.info:eu-repo/semantics/publishedVersio

Diversity and Global Distribution of Viruses of the Western Honey Bee, Apis mellifera

Funding: This research was permitted and funded by the COLOSS network (https://coloss.org/).Peer reviewedPublisher PD

Projeto BEEHEAL: promover a saúde da abelha para uma agricultura sustentável

O 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).Ao Paulo Ventura pelo acompanhamento técnico feito ao apário no primeiro ano do projeto. O BEEHEAL é financiado por ARIMNet2 (2016) com os financiadores nacionais Instituto Nacional de Investigación y Teccnologia Agraria y alimentaria (INIA – Espanha), Agence Nationale de la recherche (ARN – France), Ministry off Agriculture & Rural Development, (MOARD – Israel) e Fundação para a Ciência e a Tecnologia (FCT – Portugal)info:eu-repo/semantics/publishedVersio

Virus Prevalence in Egg Samples Collected from Naturally Selected and Traditionally Managed Honey Bee Colonies across Europe

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 &lsquo;suppressed in ovo virus infection&rsquo; 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

CYLD Enhances Severe Listeriosis by Impairing IL-6/STAT3-Dependent Fibrin Production

The facultative intracellular bacterium Listeria monocytogenes (Lm) may cause severe infection in humans and livestock. Control of acute listeriosis is primarily dependent on innate immune responses, which are strongly regulated by NF-kappa B, and tissue protective factors including fibrin. However, molecular pathways connecting NF-kappa B and fibrin production are poorly described. Here, we investigated whether the deubiquitinating enzyme CYLD, which is an inhibitor of NF-kappa B-dependent immune responses, regulated these protective host responses in murine listeriosis. Upon high dose systemic infection, all C57BL/6 Cyld(-/-) mice survived, whereas 100% of wildtype mice succumbed due to severe liver pathology with impaired pathogen control and hemorrhage within 6 days. Upon in vitro infection with Lm, CYLD reduced NF-kappa B-dependent production of reactive oxygen species, interleukin (IL)-6 secretion, and control of bacteria in macrophages. Furthermore, Western blot analyses showed that CYLD impaired STAT3-dependent fibrin production in cultivated hepatocytes. Immunoprecipitation experiments revealed that CYLD interacted with STAT3 in the cytoplasm and strongly reduced K63-ubiquitination of STAT3 in IL-6 stimulated hepatocytes. In addition, CYLD diminished IL-6-induced STAT3 activity by reducing nuclear accumulation of phosphorylated STAT3. In vivo, CYLD also reduced hepatic STAT3 K63-ubiquitination and activation, NF-kappa B activation, IL-6 and NOX2 mRNA production as well as fibrin production in murine listeriosis. In vivo neutralization of IL-6 by anti-IL-6 antibody, STAT3 by siRNA, and fibrin by warfarin treatment, respectively, demonstrated that IL-6-induced, STAT3-mediated fibrin production significantly contributed to protection in Cyld(-/-) mice. In addition, in vivo Cyld siRNA treatment increased STAT3 phosphorylation, fibrin production, pathogen control and survival of Lm-infected WT mice illustrating that therapeutic inhibition of CYLD augments the protective NF-kappa B/IL-6/STAT3 pathway and fibrin production

Epidemiology of the Microsporidium Nosema ceranae in Four Mediterranean Countries

Nosema ceranae is a highly prevalent intracellular parasite of honey bees’ midgut worldwide. This Microsporidium was monitored during a long-term study to evaluate the infection at apiary and intra-colony levels in six apiaries in four Mediterranean countries (France, Israel, Portugal, and Spain). Parameters on colony strength, honey production, beekeeping management, and climate were also recorded. Except for São Miguel (Azores, Portugal), all apiaries were positive for N. ceranae, with the lowest prevalence in mainland France and the highest intra-colony infection in Israel. A negative correlation between intra-colony infection and colony strength was observed in Spain and mainland Portugal. In these two apiaries, the queen replacement also influenced the infection levels. The highest colony losses occurred in mainland France and Spain, although they did not correlate with the Nosema infection levels, as parasitism was low in France and high in Spain. These results suggest that both the effects and the level of N. ceranae infection depends on location and beekeeping conditions. Further studies on host-parasite coevolution, and perhaps the interactions with other pathogens and the role of honey bee genetics, could assist in understanding the difference between nosemosis disease and infection, to develop appropriate strategies for its control

Comparison of two alternative store formats using a Malmquist-type index

This paper explores the differences in performance between two groups of retailing stores that operate with different formats. The study uses a Malmquist-type index to distinguish internal inefficiencies from those associated with the group (or format) characteristics. A fundamental characteristic of the new index is to compare groups in a static setting. The study described in this paper combines the use of the Malmquist index with statistical tests. The Malmquist-type index is decomposed into sub-indexes for comparing the efficiency spread between groups and the productivity differences between the best-practice frontiers of the groups. The hypothesis tests are used to verify if the differences between groups captured by the Malmquist-type index and its components are statistically significant. There are several methods based on DEA for comparing the performance of two groups, such as the program efficiency method and the comparison of efficiency distributions using statistical hypothesis tests. The method used in this paper is compared with the existing approaches to highlight its strengths and weaknesses. The applicability of the method is illustrated with a case study that compares the performance of heavy bazaar stores (that sell electrical appliances and consumer electronics) with different formats (megastores versus superstores). The study showed that the overall performance of megastores is better due to the effect of a more productive frontier. However, the efficiency spread is larger in megastores than in superstores meaning that there is scope for efficiency improvements