de Sousa Pereira Kleber, Piot Niels, Smagghe Guy, Meeus Ivan
Faculty of Bioscience Engineering, Department of Plants and Crops, Lab of Agrozoology, Ghent University, Coupure Links 653, B-9000, Ghent, Belgium.
Parasitol Res. 2019 Feb;118(2):715-721. doi: 10.1007/s00436-018-6176-0. Epub 2019 Jan 4.
Parasites of managed bees can disrupt the colony success of the host, but also influence local bee-parasite dynamics, which is regarded as a threat for wild bees. Therapeutic measures have been suggested to improve the health of managed bees, for instance, exploiting the bees' RNA interference (RNAi) pathway to treat against viral pathogens. Gut trypanosomes are an important group of bee parasites in at least two common managed bee species, i.e., managed Apis mellifera and reared Bombus terrestris. In several trypanosomes, RNAi activity is present, while in other associated genes of RNAi, such as Dicer-like (DCL) and Argonaute (AGO), it is lost. Up to date, the ability to exploit the RNAi of gut trypanosomes of bees has remained unexplored. Here, we screened parasite genomes of two honey bee protozoa (Crithidia mellificae and Lotmaria passim) and two bumble bee protozoa (Crithidia bombi and Crithidia expoeki) for the presence of DCL and AGO proteins. For C. mellificae, we constructed a double-stranded RNA (dsRNA) targeting kinetoplastid membrane protein-11 (KMP-11) to test the RNAi potential to kill this parasite. Transfection with KMP-11 dsRNA, but also adding it to the growth medium resulted in small growth reduction of the trypanosome C. mellificae, thereby showing the limited potential to apply dsRNA therapeutics to control trypanosome infection in managed honey bee species. Within bumble bees, there seems to be no application potentials against C. bombi, as we could only retrieve non-functional DCL- and AGO-related genes within the genome of this bumble bee parasite.
养殖蜜蜂的寄生虫不仅会干扰宿主蜂群的成功发展,还会影响当地蜜蜂与寄生虫的动态关系,这被视为对野生蜜蜂的一种威胁。人们已提出一些治疗措施来改善养殖蜜蜂的健康状况,例如,利用蜜蜂的RNA干扰(RNAi)途径来对抗病毒病原体。肠道锥虫是至少两种常见养殖蜜蜂物种(即养殖的西方蜜蜂和饲养的熊蜂)中的一类重要蜜蜂寄生虫。在几种锥虫中存在RNAi活性,而在RNAi的其他相关基因(如Dicer样蛋白(DCL)和Argonaute蛋白(AGO))中则不存在。到目前为止,利用蜜蜂肠道锥虫的RNAi能力尚未得到探索。在这里,我们筛选了两种蜜蜂原生动物(蜜蜂短膜虫和洛氏蜜蜂微孢子虫)和两种熊蜂原生动物(熊蜂短膜虫和埃氏短膜虫)的寄生虫基因组中是否存在DCL和AGO蛋白。对于蜜蜂短膜虫,我们构建了靶向动质体膜蛋白-11(KMP-11)的双链RNA(dsRNA),以测试其杀死这种寄生虫的RNAi潜力。用KMP-11 dsRNA转染,以及将其添加到生长培养基中,均导致蜜蜂短膜虫的生长略有减少,从而表明应用dsRNA疗法控制养殖蜜蜂物种中锥虫感染的潜力有限。在熊蜂中,似乎没有针对熊蜂短膜虫的应用潜力,因为我们只能在这种熊蜂寄生虫的基因组中检索到无功能的DCL和AGO相关基因。