Sacbrood Virus Research Articles

Colony-Level Effects of Amygdalin on Honeybees and Their Microbes.

Simple SummaryNectar compounds have the potential to affect microbial communities and pollinator immunity. Here, we investigated how the almond compound, amygdalin, influences the microbial community of the western honeybee. Using RNA sequencing technology to count microbial reads and bee gene transcripts, we show relatively no large change of bacterial counts, fungal counts or bee transcripts due to amygdalin treatment at the colony level. Larger fluctuations, perhaps due to amygdalin, were observed for pathogenic viruses and the pathogen Lotmaria passim; however, these changes could have been seasonal. Overall, amygdalin consumption at field-relevant, colony-levels may not have a large impact on bee symbionts or immune gene expression.Amygdalin, a cyanogenic glycoside, is found in the nectar and pollen of almond trees, as well as in a variety of other crops, such as cherries, nectarines, apples and others. It is inevitable that western honeybees (Apis mellifera) consistently consume amygdalin during almond pollination season because almond crops are almost exclusively pollinated by honeybees. This study tests the effects of a field-relevant concentration of amygdalin on honeybee microbes and the activities of key honeybee genes. We executed a two-month field trial providing sucrose solutions with or without amygdalin ad libitum to free-flying honeybee colonies. We collected adult worker bees at four time points and used RNA sequencing technology and our HoloBee database to assess global changes in microbes and honeybee transcripts. Our hypothesis was that amygdalin will negatively affect bee microbes and possibly immune gene regulation. Using a log2 fold-change cutoff at two and intraday comparisons, we show no large change of bacterial counts, fungal counts or key bee immune gene transcripts, due to amygdalin treatment in relation to the control. However, relatively large titer decreases in the amygdalin treatment relative to the control were found for several viruses. Chronic bee paralysis virus levels had a sharp decrease (−14.4) with titers then remaining less than the control, Black queen cell virus titers were lower at three time points (<−2) and Deformed wing virus titers were lower at two time points (<−6) in amygdalin-fed compared to sucrose-fed colonies. Titers of Lotmaria passim were lower in the treatment group at three of the four dates (<−4). In contrast, Sacbrood virus had two dates with relative increases in its titers (>2). Overall, viral titers appeared to fluctuate more so than bacteria, as observed by highly inconstant patterns between treatment and control and throughout the season. Our results suggest that amygdalin consumption may reduce several honeybee viruses without affecting other microbes or colony-level expression of immune genes.

Viral impacts on honey bee populations: A review.

Honey bee is vital for pollination and ecological services, boosting crops productivity in terms of quality and quantity and production of colony products: wax, royal jelly, bee venom, honey, pollen and propolis. Honey bees are most important plant pollinators and almost one third of diet depends on bee’s pollination, worth billions of dollars. Hence the role that honey bees have in environment and their economic importance in food production, their health is of dominant significance. Honey bees can be infected by various pathogens like: viruses, bacteria, fungi, or infested by parasitic mites. At least more than 20 viruses have been identified to infect honey bees worldwide, generally from Dicistroviridae as well as Iflaviridae families, like ABPV (Acute Bee Paralysis Virus), BQCV (Black Queen Cell Virus), KBV (Kashmir Bee Virus), SBV (Sacbrood Virus), CBPV (Chronic bee paralysis virus), SBPV (Slow Bee Paralysis Virus) along with IAPV (Israeli acute paralysis virus), and DWV (Deformed Wing Virus) are prominent and cause infections harmful for honey bee colonies health. This issue about honey bee viruses demonstrates remarkably how diverse this field is, and considerable work has to be done to get a comprehensive interpretation of the bee virology.

Identification of Immune Response to Sacbrood Virus Infection in Apis cerana Under Natural Condition.

Chinese sacbrood virus (CSBV) is a serious threat to eastern honeybees (Apis cerana), especially larvae. However, the pathological mechanism of this deadly disease remains unclear. Here, we employed mRNA and small RNA (sRNA) transcriptome approach to investigate the microRNAs (miRNAs) and small interfering RNAs (siRNAs) expression changes of A. cerana larvae infected with CSBV under natural condition. We found that serine proteases involved in immune response were down-regulated, while the expression of siRNAs targeted to serine proteases were up-regulated. In addition, CSBV infection also affected the expression of larvae cuticle proteins such as larval cuticle proteins A1A and A3A, resulting in increased susceptibility to CSBV infection. Together, our results provide insights into sRNAs that they are likely to be involved in regulating honeybee immune response.

Egg transcriptome profile responds to maternal virus infection in honey bees, Apis mellifera.

Trans-generational disease effects include vertical pathogen transmission but also immune priming to enhance offspring immunity. Accordingly, the survival consequences of maternal virus infection can vary and its molecular consequences during early development are poorly understood. The honey bee queen is long-lived and represents the central hub for vertical virus transmission as the sole reproductive individual in her colony. Even though virus symptoms in queens are mild, viral infection may have severe consequences for the offspring. Thus, transcriptome patterns during early developmental are predicted to respond to maternal virus infection. To test this hypothesis, gene expression patterns were compared among pooled honey bee eggs laid by queens that were either infected with Deformed wing virus (DWV1), Sacbrood virus (SBV2), both viruses (DWV and SBV), or no virus. Whole transcriptome analyses revealed significant expression differences of a few genes, some of which have hitherto no known function. Despite the paucity of single gene effects, functional enrichment analyses revealed numerous biological processes in the embryos to be affected by virus infection. Effects on several regulatory pathways were consistent with maternal responses to virus infection and correlated with responses to DWV and SBV in honey bee larvae and pupae. Overall, effects on egg transcriptome patterns were specific to each virus and the results of dual-infection samples suggested synergistic effects of DWV and SBV. We interpret our results as consequences of maternal infections. Thus, this first study to document and characterize virus-associated changes in the transcriptome of honey bee eggs represents an important contribution to understanding trans-generational virus effects, although more in-depth studies are needed to understand the detailed mechanisms of how viruses affect honey bee embryos.

Longitudinal monitoring of honey bee colonies reveals dynamic nature of virus abundance and indicates a negative impact of Lake Sinai virus 2 on colony health.

Honey bees (Apis mellifera) are important pollinators of plants, including those that produce nut, fruit, and vegetable crops. Therefore, high annual losses of managed honey bee colonies in the United States and many other countries threaten global agriculture. Honey bee colony deaths have been associated with multiple abiotic and biotic factors, including pathogens, but the impact of virus infections on honey bee colony population size and survival are not well understood. To further investigate seasonal patterns of pathogen presence and abundance and the impact of viruses on honey bee colony health, commercially managed colonies involved in the 2016 California almond pollination event were monitored for one year. At each sample date, colony health and pathogen burden were assessed. Data from this 50-colony cohort study illustrate the dynamic nature of honey bee colony health and the temporal patterns of virus infection. Black queen cell virus, deformed wing virus, sacbrood virus, and the Lake Sinai viruses were the most readily detected viruses in honey bee samples obtained throughout the year. Analyses of virus prevalence and abundance revealed pathogen-specific trends including the overall increase in deformed wing virus abundance from summer to fall, while the levels of Lake Sinai virus 2 (LSV2) decreased over the same time period. Though virus prevalence and abundance varied in individual colonies, analyses of the overall trends reveal correlation with sample date. Total virus abundance increased from November 2015 (post-honey harvest) to the end of the almond pollination event in March 2016, which coincides with spring increase in colony population size. Peak total virus abundance occurred in late fall (August and October 2016), which correlated with the time period when the majority of colonies died. Honey bee colonies with larger populations harbored less LSV2 than weaker colonies with smaller populations, suggesting an inverse relationship between colony health and LSV2 abundance. Together, data from this and other longitudinal studies at the colony level are forming a better understanding of the impact of viruses on honey bee colony losses.

Phylogenetic analysis of sacbrood virus structural polyprotein and non-structural RNA dependent RNA polymerase gene: Differences in Turkish strains

Sacbrood virus (SBV) is one of the most damaging viruses in honey bee colonies. Genetic differences among sacbrood viruses detected in honey bees in different locales have been reported in previous studies. The aim of this study was to construct phylogenetic trees based on the structural polyprotein and non-structural RNA dependent RNA polymerase gene regions and to make a molecular characterization of the Tur/Bur/Sac01 and Tur/Bur/Sac02 strains identified in Apis mellifera in Turkey. As a result of the study, the tree based on the structural polyprotein region separated into four lineages: Tur/Bur/Sac01 and Tur/Bur/Sac02 were in the same branch as the Turkish sacbrood virus strains identified in previous studies and formed the Turkish clade. Strains isolated from adjacent geographical areas were in the same clade in this tree. The phylogenetic tree based on the non-structural RNA dependent RNA polymerase gene region divides into two main branches, reflecting host affiliation: Apis cerana and A. mellifera. Strains formed clusters based on their geographic distribution and host affiliation. The Tur/Bur/Sac01 and Tur/Bur/Sac02 strains formed a separate cluster among the European strains. Sacbrood viruses from Turkey were genetically different from SBV strains detected in other countries and in A. cerana.

Nationwide Screening for Bee Viruses and Parasites in Belgian Honey Bees.

The health of honey bees is threatened by multiple factors, including viruses and parasites. We screened 557 honey bee (Apis mellifera) colonies from 155 beekeepers distributed all over Belgium to determine the prevalence of seven widespread viruses and two parasites (Varroa sp. and Nosema sp.). Deformed wing virus B (DWV-B), black queen cell virus (BQCV), and sacbrood virus (SBV) were highly prevalent and detected by real-time RT-PCR in more than 95% of the colonies. Acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV) and deformed wing virus A (DWV-A) were prevalent to a lower extent (between 18 and 29%). Most viruses were only present at low or moderate viral loads. Nevertheless, about 50% of the colonies harbored at least one virus at high viral load (>107 genome copies/bee). Varroa mites and Nosema sp. were found in 81.5% and 59.7% of the honey bee colonies, respectively, and all Nosema were identified as Nosema ceranae by real time PCR. Interestingly, we found a significant correlation between the number of Varroa mites and DWV-B viral load. To determine the combined effect of these and other factors on honey bee health in Belgium, a follow up of colonies over multiple years is necessary.

Identification of a Novel Picorna-like Virus in Coconut Rhinoceros Beetles (Oryctes rhinoceros)

A novel Picorna-like virus, tentatively named Oryctes rhinoceros Picorna-like virus 1 (OrPV1), was identified in coconut rhinoceros beetle (Oryctes rhinoceros) larvae in Taiwan. The complete genome sequence consisted of 9,665 nucleotides with a polyA tail and included one open reading frame. Conserved structural domains such as Picornavirus capsid protein, RNA helicase, Peptidase and RNA-dependent RNA polymerase (RdRp) were identified through Pfam domain searches. The genome shares approximately 27-28% identity with other unclassified Picornavirales that infect honey bees (Darwin bee virus 2, Bundaberg bee virus 5, and Sacbrood virus) and a recently reported virus from Asian lady beetle (Harmonia axyridis virus 1). We did not detect this virus in any other geographical populations of O. rhinoceros collected from the South Pacific Islands and the Philippines. Analysis of the deduced RdRp amino acid sequences showed that the virus clustered with other Picorna-like viruses and separated from other members of family Dicistroviridae and Iflaviridae.

Screening and Validation of Reference Genes for RT-qPCR Under Different Honey Bee Viral Infections and dsRNA Treatment.

Honey bee viruses are one of the most important pathogens that have contributed to the decrease in honey bee colony health. To analyze the infection dynamics of honey bee viruses, quantification of viral gene expression by RT-qPCR is necessary. However, suitable reference genes have not been reported from viral and RNAi studies of honey bee. Here, we evaluated the expression of 11 common reference genes (ache2, rps18, β-actin, tbp, tif, rpl32, gadph, ubc, α-tubulin, rpl14, and rpsa) from Apis mellifera (Am) and Apis cerana (Ac) under Israeli acute paralysis virus (IAPV), chronic bee paralysis virus (CBPV), and Chinese sacbrood virus (CSBV) infection as well as dsRNA-PGRP-SA treatment, and we confirmed their validation by evaluating the levels of the defensin 1 and prophenoloxidase (ppo) genes during viral infection. Our results showed that the expression of selected genes varied under different viral infections. ache2, rps18, β-actin, tbp, and tif can be used to normalize expression levels in Apis mellifera under IAPV infection, while the combination of actin and tif is suitable for CBPV-infected experiments. The combination of rpl14, tif, rpsa, ubc, and ache2 as well as more reference genes is suitable for CSBV treatment in Apis cerana. Rpl14, tif, rps18, ubc, and α-tubulin were the most stable reference genes under dsRNA treatment in Apis mellifera. Furthermore, the geNorm and NormFinder algorithms showed that tif was the best suitable reference gene for these four treatments. This study screened and validated suitable reference genes for the quantification of viral levels in honey bee, as well as for RNAi experiments.

Impact of Sacbrood Virus on Larval Microbiome of Apis mellifera and Apis cerana.

In this study, we examined the impact of Sacbrood virus (SBV), the cause of larval honeybee (Apis mellifera) death, producing a liquefied a larva sac, on the gut bacterial communities on two larval honeybee species, Apis mellifera and Apis cerana. SBV was added into a worker jelly food mixture and bee larvae were grafted into each of the treatment groups for 24 h before DNA/RNA extraction. Confirmation of SBV infection was achieved using quantitative reverse transcription polymerase chain reaction (RT-qPCR) and visual symptomology. The 16S rDNA was sequenced by Illumina sequencing. The results showed the larvae were infected with SBV. The gut communities of infected A. cerana larvae exhibited a dramatic change compared with A. mellifera. In A. mellifera larvae, the Illumina sequencing revealed the proportion of Gilliamella, Snodgrassella and Fructobacillus was not significantly different, whereas in A. cerana, Gilliamella was significantly decreased (from 35.54% to 2.96%), however, with significant increase in Snodgrassella and Fructobacillus. The possibility of cross-infection should be further investigated.

Comparative analysis of viruses in four bee species collected from agricultural, urban, and natural landscapes.

Managed honey bees (Apis mellifera L.) and wild bees provide critical ecological services that shape and sustain natural, agricultural, and urban landscapes. In recent years, declines in bee populations have highlighted the importance of the pollination services they provide and the need for more research into the reasons for global bee losses. Several stressors cause declining populations of managed and wild bee species such as habitat degradation, pesticide exposure, and pathogens. Viruses, which have been implicated as a key stressor, are able to infect a wide range of species and can be transmitted both intra- and inter-specifically from infected bee species to uninfected bee species via vertical (from parent to offspring) and/or horizontal (between individuals via direct or indirect contact) transmission. To explore how viruses spread both intra- and inter-specifically within a community, we examined the impact of management, landscape type, and bee species on the transmission of four common viruses in Nebraska: Deformed wing virus (DWV), Israeli acute paralysis virus (IAPV), Black queen cell virus (BQCV), and Sacbrood virus (SBV). Results indicated the prevalence of viruses is significantly affected (P < 0.005) by bee species, virus type, and season, but not by landscape or year (P = 0.290 and 0.065 respectively). The higher prevalence of DWV detected across bee species (10.4% on Apis mellifera, 5.3% on Bombus impatiens, 6.1% on Bombus griseocollis, and 22.44% on Halictus ligatus) and seasons (10.8% in early-mid summer and 11.4% in late summer) may indicate a higher risk of interspecific transmission of DWV. However, IAPV was predominately detected in Halictus ligatus (20.7%) and in late season collections (28.1%), which may suggest species-specific susceptibility and seasonal trends in infection rates associated with different virus types. However, there were limited detections of SBV and BQCV in bees collected during both sampling periods, indicating SBV and BQCV may be less prevalent among bee communities in this area.

The investigation of Israil acute bee paralysis virus, sacbrood virus, kashmir bee virus and chronic bee paralysis virus in honeybees (Apis mellifera)

The investigation of Israil acute bee paralysis virus, sacbrood virus, kashmir bee virus and chronic bee paralysis virus in honeybees (Apis mellifera)

Tolerance of Honey Bees to Varroa Mite in the Absence of Deformed Wing Virus.

The global spread of the parasitic mite Varroa destructor has emphasized the significance of viruses as pathogens of honey bee (Apis mellifera) populations. In particular, the association of deformed wing virus (DWV) with V. destructor and its devastating effect on honey bee colonies has led to that virus now becoming one of the most well-studied insect viruses. However, there has been no opportunity to examine the effects of Varroa mites without the influence of DWV. In Papua New Guinea (PNG), the sister species, V. jacobsoni, has emerged through a host-shift to reproduce on the local A. mellifera population. After initial colony losses, beekeepers have maintained colonies without chemicals for more than a decade, suggesting that this bee population has an unknown mite tolerance mechanism. Using high throughput sequencing (HTS) and target PCR detection, we investigated whether the viral landscape of the PNG honey bee population is the underlying factor responsible for mite tolerance. We found A. mellifera and A. cerana from PNG and nearby Solomon Islands were predominantly infected by sacbrood virus (SBV), black queen cell virus (BQCV) and Lake Sinai viruses (LSV), with no evidence for any DWV strains. V. jacobsoni was infected by several viral homologs to recently discovered V. destructor viruses, but Varroa jacobsoni rhabdovirus-1 (ARV-1 homolog) was the only virus detected in both mites and honey bees. We conclude from these findings that A. mellifera in PNG may tolerate V. jacobsoni because the damage from parasitism is significantly reduced without DWV. This study also provides further evidence that DWV does not exist as a covert infection in all honey bee populations, and remaining free of this serious viral pathogen can have important implications for bee health outcomes in the face of Varroa.

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

In the past centuries, viruses have benefited from globalization to spread across the globe, infecting new host species and populations. A growing number of viruses have been documented in the western honey bee, Apis mellifera. Several of these contribute significantly to honey bee colony losses. This review synthetizes the knowledge of the diversity and distribution of honey-bee-infecting viruses, including recent data from high-throughput sequencing (HTS). After presenting the diversity of viruses and their corresponding symptoms, we surveyed the scientific literature for the prevalence of these pathogens across the globe. The geographical distribution shows that the most prevalent viruses (deformed wing virus, sacbrood virus, black queen cell virus and acute paralysis complex) are also the most widely distributed. We discuss the ecological drivers that influence the distribution of these pathogens in worldwide honey bee populations. Besides the natural transmission routes and the resulting temporal dynamics, global trade contributes to their dissemination. As recent evidence shows that these viruses are often multihost pathogens, their spread is a risk for both the beekeeping industry and the pollination services provided by managed and wild pollinators.

Development of a multiplex RT-PCR assay for the routine detection of seven RNA viruses in Apis mellifera

Colony losses in apiaries are frequently one of the most important problems in beekeeping. Colony loss is multifactorial with many reported disorders, Colony Collapse Disorder (CCD), is an increasingly recognised phenomenon which is thought to be caused by many pathogens, including viruses. The aim of this study was to develop a multiplex RT-PCR (mRT-PCR) test to obtain faster results in routine diagnostic laboratories for seven crucial bee viruses. Specific primers for seven RNA viruses, including Israeli acute bee paralysis virus (IAPV), deformed wing virus (DWV), sacbrood virus (SBV), acute bee paralysis virus (ABPV), black queen cell virus (BQCV), kashmir bee virus (KBV) and chronic paralysis virus (CBPV), were used for testing procedure. The mRT-PCR assay can amplify seven plasmid DNA fragments from the pooled viral genomes and it was shown to be sensitive because virus copy numbers were detected to be 104 copies/μl when log10 serial dilutions were performed for the optimized mRT- PCR method. It is concluded that, mRT-PCR test can be used in routine analysis because this assay can perform specific, sensitive and reliable results also achieves economic gains and time due to detecting seven viral agents simultaneously.

Visualizing Sacbrood Virus of Honey Bees via Transformation and Coupling with Enhanced Green Fluorescent Protein

Sacbrood virus (SBV) of honey bees is a picornavirus in the genus Iflavirus. Given its relatively small and simple genome structure, single positive-strand RNA with only one ORF, cloning the full genomic sequence is not difficult. However, adding nonsynonymous mutations to the bee iflavirus clone is difficult because of the lack of information about the viral protein processes. Furthermore, the addition of a reporter gene to the clones has never been accomplished. In preliminary trials, we found that the site between 3′ untranslated region (UTR) and poly(A) can retain added sequences. We added enhanced green fluorescent protein (EGFP) expression at this site, creating a SBV clone with an expression tag that does not affect virus genes. An intergenic region internal ribosome entry site (IRES) from Black queen cell virus (BQCV) was inserted to initiate EGFP expression. The SBV-IRES-EGFP clone successfully infected Apis cerana and Apis mellifera, and in A. cerana larvae, it was isolated and passaged using oral inoculation. The inoculated larvae had higher mortality and the dead larvae showed sacbrood symptoms. The added IRES-EGFP remained in the clone through multiple passages and expressed the expected EGFP in all infected bees. We demonstrated the ability to add gene sequences in the site between 3′-UTR and poly(A) in SBV and the potential to do so in other bee iflaviruses; however, further investigations of the mechanisms are needed. A clone with a desired protein expression reporter will be a valuable tool in bee virus studies.

Occurrence of honey bee-associated pathogens in Varroa-free pollinator communities

Australia remains the last significant land mass free of Varroa, a parasitic mite which has caused dramatic honey bee (Apis mellifera) colony losses across the globe, due to its association with the pathogenic deformed wing virus (DWV). As such, Australia continues to maintain relatively healthy honey bee populations, despite recent work showing apiaries harbor a surprisingly high prevalence of microbial pathogens. We sought to determine the prevalence of these microbial pathogens in honey bees and native pollinators actively co-foraging on mass flowering crops and to understand the extent to which they may be shared between taxa. We found high prevalences of black queen cell virus (BQCV) and sacbrood virus (SBV) in the honey bees (88% and 41% respectively), and correspondingly, these were the most common honey bee pathogens detected in native pollinator taxa, albeit at much lower prevalence; the maximum prevalence for any pathogen in a native pollinator group was 24% (BQCV in Halictidae spp.). The viral pathogens Israeli acute paralysis virus and Lake Sinai viruses 1 and 2, and the fungal parasites Nosema apis and Nosema ceranae, were only rarely detected. Phylogenetic analyses of the most common pathogens revealed similar genotypes circulating between species. Our data suggest that, in Australian orchards, pathogen prevalence in honey bees is a good predictor of pathogen prevalence in native pollinators, which raises concerns about how the viral landscape may change in native taxa if, or when, Varroa arrives.

Relative Abundance and Status of Arthropod Pests and Diseases of Honey Bees at South Ari District of South Western Ethiopia

Aim: To determine the species composition, relative abundance and status of honey bee pests and diseases in South Ari district of South Omo Zone of Southern Western Ethiopia.&#x0D; Study Design: Four major honey producing peasant associations (PAs) representing diverse ecological zones were selected purposively, while the sub-localities, villages and beekeepers were selected randomly using a nested design.&#x0D; Place and Duration of Study: Survey was conducted between 1, August – 30, December 2019 with the interval of one month in major honey producing peasant associations representing diverse ecological zones of Ari district of South Western Ethiopia.&#x0D; Methodology: The assessment on abundance and status was made from of half kilogram of fresh and dry honey comb samples that were taken from 720 farmer’s hives of four peasant associations using taxonomic key of books related to arthropod pests and diseases of honey bee.&#x0D; Results: Five species of predators and parasitoids belonging to four orders were recorded. Of these 76.63% accounts for small hive beetles, followed ants (38.31%) and wasps (19.92%). Besides, six type’s diseases of honey bee caused by different pathogens were recorded. Of which, Chalk brood, other virosis, European foulbrood and American foulbrood, respectively were the most frequently occurring. In terms of the status, the diseases of honey bee were in the following orders, i.e., Chalk brood &gt; other virosis &gt; European foulbrood &gt; American foulbrood &gt; Sac Brood Virus, while their pests were in the following orders, i.e., Small hive beetles &gt; Ants &gt; Wasps &gt; Wax moths &gt; Spiders.&#x0D; Conclusion: Arthropod pests and diseases were the most important constraints of honey colony and honey production in the study areas. Thus, it is important to design and implement effective management strategies to reduce loss of honey colony and honey production in the study area.

The impact of lowbush blueberry (Vaccinium angustifolium Ait.) and cranberry (Vaccinium macrocarpon Ait.) pollination on honey bee (Apis mellifera L.) colony health status.

Commercial lowbush blueberry (Vaccinium angustifolium Ait.) and cranberry (Vaccinium macrocarpon Ait.) crops benefit from the presence of honey bee (Apis mellifera L.) for pollination. Unfortunately, beekeepers are observing negative impacts of pollination services on honey bee colonies. In this study, we investigated three beekeeping management strategies (MS) and measured their impact on honey bee colony health and development. Experimental groups (five colonies/MS) were: A) Control farmland honey producing MS (control MS); B) Blueberry pollination MS (blueberry MS); C) Cranberry pollination MS (cranberry MS) and D) Double pollination MS, blueberry followed by cranberry (double MS). Our goals were to 1) compare floral abundance and attractiveness of foraging areas to honey bees between apiaries using a Geographic Information System, and 2) compare honey bee colony health status and population development between MS during a complete beekeeping season. Our results show significantly lower floral abundance and honey bee attractiveness of foraging areas during cranberry pollination compared to the other environments. The blueberry pollination site seemed to significantly reduce brood population in the colonies who provided those services (blueberry MS and double MS). The cranberry pollination site seemed to significantly reduce colony weight gain (cranberry MS and double MS) and induce a significantly higher winter mortality rate (cranberry MS). We also measured significantly higher levels of Black queen cell virus and Sacbrood virus in the MS providing cranberry pollination (cranberry MS and double MS).

Resistance of native honey bees from Rhodope Mountains and lowland regions of Bulgaria to Nosema ceranae and viral pathogens

The Western honey bee (Apis mellifera L., Hymenoptera: Apidae) is a species of fundamental economic, agricultural and environmental importance. The aim of this study was to compare the prevalence of some parasitic and viral pathogens in local honey bees from the Rodope Mountains and plain regions. To achieve this goal, molecular screening for two of the most distributed Nosema spp. and molecular identification of six honey bee viruses – Deformed wing virus (DWV), Acute bee paralysis virus (ABPV), Chronic bee paralysis virus (CBPV), Sacbrood virus (SBV), Kashmir bee virus (KBV), and Black queen cell virus (BQCV) was performed. Molecular analysis was carried out on 168 honey bee samples from apiaries situated in three different parts of the country where a mix of different honey bee subspecies were reared. In South Bulgaria (the Rhodope Mountains), a local honey bee called Apis mellifera rodopica (a local ecotype of A. m. macedonica) was bred, while in the other two regions (plains) different introduced subspecies existed. The results showed that the samples from the lowland regions in the country were outlined with the highest prevalence (70.5%) of N. ceranae, while those from the mountainous parts had the lowest rate (5.2%). Four of the honey bee viruses were identified – DWV (10/5.9%), followed by SBV (6/3.6%) and ABPV (2/1.2%), and one case of BQCV. In conclusion, the local honey bee A. m. rodopica (despite the higher number of samples) has shown lower prevalence of both nosemosis and viral infections. Therefore, this honey bee has to be preserved as a part of the national biodiversity.