Effect of api-bioxal® and apiherb® treatments against Nosema ceranae infection in Apis mellifera investigated by two qPCR methods

Nosema ceranae is a worldwide distributed midgut parasite of western honey bees, leading to dwindling colonies and their collapse. As a treatment, only fumagillin is available, causing issues like resistance and hampered bee physiology. This study aimed to evaluate ApiHerb® and Api-Bioxal® as treatments against N. ceranae. The efficacy was tested using two qPCR methods based on the 16S rRNA and Hsp70 genes. In addition, these methods were compared for their aptitude for the quantification of the infection. For this, 19 colonies were selected based on the presence of N. ceranae infections. The colonies were divided into three groups: treated with ApiHerb, Api-Bioxal with previous queen caging and an untreated control. All colonies were sampled pre- and post-treatment. The bees were analyzed individually and in duplicate with both qPCR methods. All bees in the pre-treatment tested positive for N. ceranae. Both treatments reduced the abundance of N. ceranae, but ApiHerb also decreased the prevalence of infected bees. Analysis with the 16S rRNA method resulted in several orders of magnitude more copies than analysis with the Hsp70 method. We conclude that both products are suitable candidates for N. ceranae treatment. From our analysis, the qPCR method based on the Hsp70 gene results as more apt for the exact quantification of N. ceranae as is needed for the development of veterinary medicinal products

Microbiome network in the pelagic and benthic offshore systems of the northern Adriatic Sea (Mediterranean Sea)

Because of their recognized global importance, there is now the urgent need to map diversity and distribution patterns of marine microbial communities. Even if available studies provided some advances in the understanding the biogeographical patterns of marine microbiomes at the global scale, their degree of plasticity at the local scale it is still underexplored, and functional implications still need to be dissected. In this scenario here we provide a synoptical study on the microbiomes of the water column and surface sediments from 19 sites in a 130 km2 area located 13.5 km afar from the coast in the North-Western Adriatic Sea (Italy), providing the finest-scale mapping of marine microbiomes in the Mediterranean Sea. Pelagic and benthic microbiomes in the study area showed sector specific-patterns and distinct assemblage structures, corresponding to specific variations in the microbiome network structure. While maintaining a balanced structure in terms of potential ecosystem services (e.g., hydrocarbon degradation and nutrient cycling), sector-specific patterns of over-abundant modules-and taxa-were defined, with the South sector (the closest to the coast) characterized by microbial groups of terrestrial origins, both in the pelagic and the benthic realms. By the granular assessment of the marine microbiome changes at the local scale, we have been able to describe, to our knowledge at the first time, the integration of terrestrial microorganisms in the marine microbiome networks, as a possible natural process characterizing eutrophic coastal area. This raises the question about the biological threshold for terrestrial microorganisms to be admitted in the marine microbiome networks, without altering the ecological balance

Conservation status of Apis mellifera ruttneri inferred from whole genome sequences

Apis mellifera ruttneri is the native honey bee subspecies from the Maltese islands (Malta, Comino and Gozo). This African-lineage subspecies is adapted to the harsh environmental conditions and to the limited forage season of these islands. For many years, it was considered highly hybridized, due to the large and sustained importation of foreign subspecies, especially A. m. ligustica, perceived by many beekeepers as superior or for commercial reasons. Recent studies based on morphometry and mitochondrial DNA have suggested the occurrence of A. m. ruttneri, particularly in the main island. Conservation initiatives and a breeding program have been developed, but to date its conservation status is not fully known. As part of the MEDIBEES project, here, over 50 A. m. ruttneri georeferenced colonies collected from across Malta, as well as 50 several reference subspecies (A. m. ligustica, A. m. siciliana, A. m. iberiensis, A. m. sahariensis, and A. m. intermissa) were whole genome sequenced. The population structure and admixture were assessed from genome-wide single nucleotide polymorphisms using model and distance based-methods. The results show varying levels of admixture proportions of A. m. ruttneri with C-lineage honey bees across Malta but also a shared ancestry with the honey bees of north Africa, consistent with the putative natural colonization from Africa in ancient times.info:eu-repo/semantics/publishedVersio

MEDIBEES: Monitoring the Mediterranean honey bee subspecies and their resilience to climate change for the improvement of sustainable agro-ecosystems

Beekeeping provides livelihood for hundreds of thousands of beekeepers in the Mediterranean area. This activity relies on a number of different indigenous Apis mellifera subspecies, adapted to the very diverse and harsh conditions of the region. Climate change is expected to increase the stress factors affecting bees, especially in this region, reducing both pollination efficiency and production potential. Unfortunately, our ability to address this problem is limited by the incomplete knowledge of the natural adaptation mechanisms developed by the different subspecies. In order to increase the knowledge base for future selection programs to improve bee populations for environmental changes, the MEDIBEES project is being developed. It includes 9 partners from 8 Mediterranean countries on all three shores of the Mediterranean, covering 10 local A. mellifera subspecies, which represents a remarkable though understudied proportion of the species genetic diversity. The project aims to: a) unravel the differential genetic background of Mediterranean subspecies, b) understand their adaptation to local conditions, and c) characterize their resistance to climate change. To achieve the objectives, colonies belonging to the local subspecies will be studied phenotypically to determine their behavior under environmental conditions, covering survival, sensitivity to pests/pathogens, behavior, physiology and reproduction which will be completed by gene expression and transcriptomic assays. In addition, complete genomes of field and laboratory samples will be sequenced to find genes putatively involved in adaptation and to develop new genetic tools to characterize honey bee populations according to their resistance to environmental stress factors. This effort will encourage the use of local subspecies, to make them more attractive and avoid importing foreign breeds, and will lay the foundation for future selection programs. Besides, the valorization of honey by both promoting its use and developing quality labels, and the evaluation of beekeeping by-products as modifiers of soil fertility and biota are also approached to help the beekeepers improve the sustainability of their farms in an economical and environmental sound manner.Medibees part of the PRIMA programme supported by the European Unioninfo:eu-repo/semantics/publishedVersio

Beekeeping in the Mediterranean: a medibees survey on practices, trends and challenges

MEDIBEES is a PRIMA funded project that aims to identify honeybee colonies and subspecies that are most resilient to climate change and common pathogens throughout the Mediterranean region. Honeybees are critical to sustainable agricultural systems and the project aims to contribute to sustainability in the context of the increased desertification of the region due to climate change. As part of this project an intensive investigation of beekeeping was carried out within the consortium, with the objectives of 1) understanding the main practices in place, 2) identifying trends and challenges faced by beekeepers. This would help guide the consortium in framing research questions that are rooted in the requirements of the stakeholders. A questionnaire was therefore developed and circulated among beekeepers from Algeria, Egypt, Italy, Jordan, Lebanon, Malta, Portugal, Spain and Turkey in late June 2021. To date, over 1300 detailed responses have been collected. This study has yielded an important first profile of the nature of the apicultural industry for many of these participating countries. In this presentation, a description of the socio-demographics and the diverse management practice in place among beekeepers across the Mediterranean will be presented. In addition, important first-time data on the distribution of hives, the main sources of losses including current biotic and abiotic threats, the characteristics and ecological trends of both native and introduced honey bee subspecies in the different geographical regions will be described as experienced by the major stakeholders ie beekeepers. Finally, notably and overwhelmingly, beekeepers from the Mediterranean region list changes in weather patterns as the major factor affecting colony losses and profitability. This was followed closely by indiscriminant use of agricultural insecticides and urbanisation.info:eu-repo/semantics/publishedVersio

Main practices, challenges and expected solution identified in the Mediterranean beekeeping industry

Globally honeybees are threatened by a plethora of biotic and abiotic stressors. One factor of great concern is climate change due to its direct impact on wild and agricultural ecosystems. Indeed beekeeping, with its symbiotic interdependence with vegetation, is an industry that will be strongly affected by climate change. Although the Mediterranean is a region that is highly vulnerable to climate change, no study has comprehensively analysed the possible effects on honeybees. The MEDIBEES project aims to remedy this situation by studying the honeybee subspecies native to the Mediterranean region and their resilience to climate change, in an attempt to safeguard the beekeeping sector. In this framework, the following approaches were adopted to determine the common Practices, Challenges and Problems in the region and to identify scientific gaps and solutions expected by beekeepers: • Review of the effects of biotic and abiotic stressors on honeybee colonies as well as legislation in the MEDIBEES consortium to identify common threats. • Study of current challenges, priority areas considered by beekeepers as important to survive as an industry, beekeeping practices and existing resources in each country to determine common trends and challenges across the Mediterranean. • Analysis of the responses to a questionnaire distributed among 1160 beekeepers to obtain information on stakeholder practices, challenges faced and the main problems encountered. In this way, a profile of beekeeping activity and beekeepers in the region was obtained. The main challenges and problems identified were: a) changes in weather patterns that could be affect colony dynamics, production losses and increasing mortality; b) of incidence of pathogens in the area (Varroa destructor, Nosema ceranae, SBV and DWV) and the lack of training programmes to control them; c) agricultural insecticides and acaricide residues. As main areas of interest, beekeepers pointed out the possible existence of adapted ecotypes, the need for training to distinguish between hybrids and purebreds, and the study of the effect that climate change will have on phenology and plant availability and on changes in the distribution areas of pathogens in the area. This project is part of the PRIMA programme supported by the European Union.info:eu-repo/semantics/publishedVersio

Treatments of honey bee colonies affected by Varroa destructor in organic apiculture

The organic honey bee productions are provided for in the EU Regulation 1804/99. Notwithstanding the beekeeping industry deserves a considerable economic importance to the whole organic market of the EU, limited action is paid in favour of the organic beekeeping. A noticeable exception to this are the huge research efforts on the disease control, which is mainly due to the lack of useful and not contaminating remedies against severe pests hitting the honey bees in the whole EU. To avoid the honey contamination with treatment residues is paramount and fosters researche on soft chemicals like oxalic acid, which is of utmost impor-tance not only in organic, but also in conventional contexts. The Authors are presenting here a synthetic review on the treatments against the varroosis with OA and on their research to optimise the use of the substance in different Italian environments

General beekeeping practices and main stressors identified by beekeepers in the Mediterranean

Beekeeping is an ecologically and economically important activity in the Mediterranean that is increasingly under threat from a combination of factors. In the MEDIBEES (Monitoring the Mediterranean honey bee subspecies and their resilience to climate change for the improvement of sustainable agro-ecosystems) PRIMA project we aim to identify honey bees that show resilience to stressors associated with climate change. An important first step in our work was to establish baseline data of practices in the region through the administering of a questionnaire on the beekeeping practices. Over 1022 responses were received from beekeepers in Algeria, Egypt, Jordan, Italy, Lebanon, Malta, Spain, Portugal and Turkey. Beekeeping was overwhelmingly male dominated (ratio 9:1) and practiced largely by the middle-aged (mainly between 40-50 years). The majority of beekeepers reported having 10-50 boxes with the exceptions of Algeria and Jordan where the majority reported keeping more than 100 hive boxes. Across the Mediterranean the Langstroth hive box was most commonly used. Despite the perception of migratory beekeeping being frequently practiced, only Lebanon and Turkey reported significant levels of transhumance (>50.0 and 70.0% of beekeepers respectively). 51.2% of beekeepers reported practicing queen rearing with the majority of these (51.4%) reporting re-queening their colonies every 2 years. Interestingly, the majority of all beekeepers agreed with the statement that their native honey bee is endangered where pesticide use, lack of adequate forage and parasite infestations such as the Varroa mite were repeatedly cited as causing the greatest losses to honey bees in general. Climate change and urbanisation were also linked with colony losses by causing habitat loss. Supplementary feeding was reported to be an integral part of beekeeping in this region, with beekeepers reporting using between 0-5kg, closely followed by 6-10 kg of additional sugars.MEDIBEES part of the PRIMA programme supported by the European Unioninfo:eu-repo/semantics/publishedVersio

Identification of genetic variants associated with Huntington's disease progression: a genome-wide association study

Background Huntington's disease is caused by a CAG repeat expansion in the huntingtin gene, HTT. Age at onset has been used as a quantitative phenotype in genetic analysis looking for Huntington's disease modifiers, but is hard to define and not always available. Therefore, we aimed to generate a novel measure of disease progression and to identify genetic markers associated with this progression measure. Methods We generated a progression score on the basis of principal component analysis of prospectively acquired longitudinal changes in motor, cognitive, and imaging measures in the 218 indivduals in the TRACK-HD cohort of Huntington's disease gene mutation carriers (data collected 2008–11). We generated a parallel progression score using data from 1773 previously genotyped participants from the European Huntington's Disease Network REGISTRY study of Huntington's disease mutation carriers (data collected 2003–13). We did a genome-wide association analyses in terms of progression for 216 TRACK-HD participants and 1773 REGISTRY participants, then a meta-analysis of these results was undertaken. Findings Longitudinal motor, cognitive, and imaging scores were correlated with each other in TRACK-HD participants, justifying use of a single, cross-domain measure of disease progression in both studies. The TRACK-HD and REGISTRY progression measures were correlated with each other (r=0·674), and with age at onset (TRACK-HD, r=0·315; REGISTRY, r=0·234). The meta-analysis of progression in TRACK-HD and REGISTRY gave a genome-wide significant signal (p=1·12 × 10−10) on chromosome 5 spanning three genes: MSH3, DHFR, and MTRNR2L2. The genes in this locus were associated with progression in TRACK-HD (MSH3 p=2·94 × 10−8 DHFR p=8·37 × 10−7 MTRNR2L2 p=2·15 × 10−9) and to a lesser extent in REGISTRY (MSH3 p=9·36 × 10−4 DHFR p=8·45 × 10−4 MTRNR2L2 p=1·20 × 10−3). The lead single nucleotide polymorphism (SNP) in TRACK-HD (rs557874766) was genome-wide significant in the meta-analysis (p=1·58 × 10−8), and encodes an aminoacid change (Pro67Ala) in MSH3. In TRACK-HD, each copy of the minor allele at this SNP was associated with a 0·4 units per year (95% CI 0·16–0·66) reduction in the rate of change of the Unified Huntington's Disease Rating Scale (UHDRS) Total Motor Score, and a reduction of 0·12 units per year (95% CI 0·06–0·18) in the rate of change of UHDRS Total Functional Capacity score. These associations remained significant after adjusting for age of onset. Interpretation The multidomain progression measure in TRACK-HD was associated with a functional variant that was genome-wide significant in our meta-analysis. The association in only 216 participants implies that the progression measure is a sensitive reflection of disease burden, that the effect size at this locus is large, or both. Knockout of Msh3 reduces somatic expansion in Huntington's disease mouse models, suggesting this mechanism as an area for future therapeutic investigation

Polymorphism of <i>16s rRNA</i> Gene: Any Effect on the Biomolecular Quantitation of the Honey Bee (<i>Apis mellifera</i> L., 1758) Pathogen <i>Nosema ceranae</i>?

The microsporidian Nosema ceranae is a severe threat to the western honey bee Apis mellifera, as it is responsible for nosemosis type C, which leads the colonies to dwindle and collapse. Infection quantification is essential to clinical and research aims. Assessment is made often with molecular assays based on rRNA genes, which are present in the N. ceranae genome as multiple and polymorphic copies. This study aims to compare two different methods of Real-Time PCR (qPCR), respectively relying on the 16S rRNA and Hsp70 genes, the first of which is described as a multiple and polymorphic gene. Young worker bees, hatched in the laboratory and artificially inoculated with N. ceranae spores, were incubated at 33 °C and subject to different treatment regimens. Samples were taken post-infection and analyzed with both qPCR methods. Compared to Hsp70, the 16S rRNA method systematically detected higher abundance. Straightforward conversion between the two methods is made impossible by erratic 16s rRNA/Hsp70 ratios. The 16s rRNA polymorphism showed an increase around the inoculated dose, where a higher prevalence of ungerminated spores was expected due to the treatment effects. The possible genetic background of that irregular distribution is discussed in detail. The polymorphic nature of 16S rRNA showed to be a limit in the infection quantification. More reliably, the N. ceranae abundance can be assessed in honey bee samples with methods based on the single-copy gene Hsp70