Salkova Delka, Balkanska Ralitsa, Shumkova Rositsa, Lazarova Stela, Radoslavov Georgi, Hristov Peter
Department of Experimental Parasitology, Institute of Experimental Morphology, Pathology and Anthropology with Museum, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria.
Department "Special Branches", Institute of Animal Science, Kostinbrod, Agricultural Academy, 1113 Sofia, Bulgaria.
Vet Sci. 2024 Aug 12;11(8):369. doi: 10.3390/vetsci11080369.
In the last few years, the isolation and amplification of DNA or RNA from the environment (eDNA/eRNA) has proven to be an alternative and non-invasive approach for molecular identification of pathogens and pests in beekeeping. We have recently demonstrated that bee pollen and bee bread represent suitable biological material for the molecular identification of viral RNA. In the present study, we extracted total RNA from different bee products (pollen, n = 25; bee bread, n = 17; and royal jelly, n = 15). All the samples were tested for the presence of six of the most common honey bee-associated 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)-using a reverse transcription polymerase chain reaction (RT-PCR). We successfully detected six records of DWV (10.5%, 6/57), four of ABPV (7.0%, 4/57), three of Israeli acute paralysis virus (IAPV) (5.3%, 3/57), and two of BQCV (3.5%, 2/57). Using ABPV primers, we also successfully detected the presence of IAPV. The obtained viral sequences were analyzed for phylogenetic relationships with the highly similar sequences (megablast) available in the GenBank database. The Bulgarian DWV isolates revealed a high homology level with strains from Syria and Turkey. Moreover, we successfully detected a DWV strain B for the first time in Bulgaria. In contrast to DWV, the ABPV isolates formed a separate clade in the phylogenetic tree. BQCV was closely grouped with Russian isolates, while Bulgarian IAPV formed its own clade and included a strain from China. In conclusion, the present study demonstrated that eRNA can be successfully used for molecular detection of honey bee-associated viruses in bee products. The method can assist the monitoring of the health status of honey bee colonies at the local, regional, and even national levels.
在过去几年中，从环境中分离和扩增DNA或RNA（环境DNA/环境RNA）已被证明是一种用于养蜂业中病原体和害虫分子鉴定的替代且非侵入性的方法。我们最近证明，蜂花粉和蜂粮是用于病毒RNA分子鉴定的合适生物材料。在本研究中，我们从不同的蜂产品（花粉，n = 25；蜂粮，n = 17；蜂王浆，n = 15）中提取了总RNA。使用逆转录聚合酶链反应（RT-PCR）对所有样品检测了六种最常见的与蜜蜂相关的病毒——残翅病毒（DWV）、急性蜜蜂麻痹病毒（ABPV）、慢性蜜蜂麻痹病毒（CBPV）、囊状幼虫病毒（SBV）、克什米尔蜜蜂病毒（KBV）和黑蜂王台病毒（BQCV）的存在情况。我们成功检测到6例DWV（10.5%，6/57）、4例ABPV（7.0%，4/57）、3例以色列急性麻痹病毒（IAPV）（5.3%，3/57）和2例BQCV（3.5%，2/57）。使用ABPV引物，我们还成功检测到了IAPV的存在。对获得的病毒序列与GenBank数据库中高度相似的序列（megablast）进行了系统发育关系分析。保加利亚的DWV分离株与来自叙利亚和土耳其的菌株显示出高度同源性。此外，我们首次在保加利亚成功检测到了DWV B株。与DWV不同，ABPV分离株在系统发育树中形成了一个单独的分支。BQCV与俄罗斯分离株紧密聚类，而保加利亚的IAPV形成了自己的分支，其中包括一个来自中国的菌株。总之，本研究表明环境RNA可成功用于蜂产品中与蜜蜂相关病毒的分子检测。该方法可协助在地方、区域甚至国家层面监测蜂群的健康状况。
