To identify viruses and compare their abundance levels in the venom glands of hymenopteran species, we conducted venom gland-specific transcriptome assemblies and analyses of 22 Aculeate bees and wasps and identified the RNA genomes of picornaviruses. Additionally, we investigated the expression patterns of viruses in the venom glands over time following capture. Honeybee-infecting viruses, including black queen cell virus (BQCV), deformed wing virus (DWV), and Israeli acute paralysis virus (IAPV), were highly expressed in the venom glands of Apis mellifera and social wasps. This finding suggests that the venoms of bees and wasps likely contain these viruses, which can be transmitted horizontally between species through their stinger usage. A. mellifera exhibited an increasing pattern of abundance levels for BQCV, DWV, IAPV, and Triatovirus, while the social wasp Vespa crabro showed increasing abundance levels of IAPV and Triatovirus over different capture periods. This suggests that the venom glands of honeybees and wasps may provide suitable conditions for active viral replication and may be an organ for virus accumulation and transmission. Some viral sequences clearly reflected the phylogeny of Aculeate species, implying host-specific virus evolution. On the other hand, other viruses exhibited unique evolutionary patterns of phylogeny, possibly caused by specific ecological interactions. Our study provides insights into the composition and evolutionary properties of viral genes in the venom glands of certain Aculeate bees and wasps, as well as the potential horizontal transmission of these viruses among bee and wasp species.
Over the course of two winters, the significant decline in honey bee populations in Korea has emerged as a major social issue. This phenomenon is expected as attributed to factors such as the failure of pest control due to the pesticide resistance of the Varroa mite. This mite can transmit some viruses that infect honey bees, and these viruses are among the primary causes of the globally occurring colony collapse disorder. Traditional diagnostic methods like (RT-)PCR and ELISA are not ideal for identifying pathogens that are newly emerging or have undergone mutations. To detect any novel or mutated viruses beyond those that have been primarily diagnosed in Korea, we introduced virome analysis technology in the field of honey bees. Employing this method with high-throughput sequencing techniques, we were able to identify all existing viruses within individual or group samples. We discovered that the Lake Sinai virus, which has been reported worldwide but not in Korea, has already significantly spread within the country. Additionally, we were able to confirm the prevalence of viruses previously reported in Korea, such as the recently dominant Black Queen Cell Virus. Through this virome analysis, we can provide foundational data for determining the direction and countermeasures for virus diagnosis.
Sacbrood virus (SBV) caused significant colony collapse in Korean Apis cerana. Considering that hygienic behavior in honey bees confers colony-level resistance against brood diseases, we utilized this trait for selecting A. cerana colonies. In addition, the brood survival rate was evaluated after colonies were SBV-inoculated. Over four selective generations, dead brood removal and brood survivorship in selected colonies were higher than those in the unselected colonies (P < 0.01, 99.3 vs. 89.9% for removal of pin-killed pupae; P < 0.01, 99.0 vs. 63.9% for removal of SBV-killed larvae; and P < 0.01, 70.0 vs. 9.2% for brood survivorship). Following SBV-inoculation, selected colonies showed an increase in the number of surviving pupae and adults, whereas unselected colonies collapsed mostly. Our results confirm the feasibility of selecting SBV-resistant A. cerana.
Sacbrood disease is a viral disease on honey bee larvae Apis cerana. Diseased larvae fail to pupae and to be dead at old larvae and pre-pupae stage. Currently, there is no remedy to control sacbrood disease. In this study we conducted to observe sacbrood disease on Apis cerana colonies from June to September, 2014 at the A. cerana apiary of NAAS, and using biological measure to treat this disease. Our study results were showed that sacbrood disease infected A. cerana colonies in all months of observation. The percentage of infected colonies was from 33.3% up to 100%. Controlling sacbrood disease by requeen measure, the percentage of recovered colonies was 57.1 % while of this by cage queen measure was only 28.6 %.
Sacbrood virus (SBV) is one of the most destructive honey bee virus. The virus causes failure to pupate and kills honey bee larvae. The infacted larvae`s color is change to brown. At the end, honey bee colony is destructed. Recently Korean Scabrood virus(KSBV) caused a great loss of Korean honey bee(Apis cerena) colonies for short period. Therefore, We need a highly rapid diagnosis method for rapid detection of KSBV.
In this study, We need amicro-scale chip-based real-time PCR system (GeneChecker®). This system was developed for rapid, specific PCR based diagnosis. This system has uncommonly fast heating and cooling system. So We was able to detecting of KSBV in Apis cerena in short time. This system needs small reaction volume(total 10ul). This volume include SsoFast™ Evagreen Supermix and serially diluted cDNA templates showed a high sensitivity of 101copies.That machine can setting each PCR stage time. A specific detection primer set (KSBV-123-F/R) was used to amplify a unique 123bp DNA fragment.
This PCR assays using serially diluted cDNA templates showed a high sensitivity of 101 copies. When applied to KSBV-positve samples, the result showed high specifity. The minimum diagnosis time was 9m 47s (30cycle). The amplied positive samples appear red fluorescent color.
This novel detection method could be used a PCR-based diagnositic tool (GeneChecker®). The results showed high sensitivity and specifity in short time. And this diagnosis method is expected to be applied to rapidly detect various pathogens.
Viruses of the honeybee, Apis mellifera L. are known to reside at low levels in colonies, typically showing no apparent signs of infection. Chronic paralysis virus(CBPV) is known to induce significant losses in honey bee colonies. The pathology is characterized by clusters of trembling, flightless, crawling bees and by individual bees, sometimes hairless, standing at the hive entrance. A minusstrand-specific RT-PCR was used to assess viral replication. This is the first report on the infection of CBPV in Korea. Using (-)RT-PCR, 27 apiaries in korea were screened for the honeybee viruses, with positive colonies being analysed for viral genetic diversity. We got 550-nt PCR product from CBPV genomic RNA. Nucleotide sequences were aligned to the complete CBPV genomic RNA sequence deposited in the GenBank database and was revealed 96%(AM-CBPV) identity, respectively. Sequence comparison with other CBPV and honeybee virus.