Non-Lethal Detection of Frog Virus 3-Like (RUK13) and Common Midwife Toad Virus-Like (PDE18) Ranaviruses in Two UK-Native Amphibian Species.
Non-Lethal Detection of Frog Virus 3-Like (RUK13) and Common Midwife Toad Virus-Like (PDE18) Ranaviruses in Two UK-Native Amphibian Species.
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DOI：
10.3390/v14122635
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发表时间：
2022-11-25
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--
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蛙病毒参与了世界范围内两栖动物的大规模死亡事件。有效的筛选，以控制这种病原体是必不可少的，但是，目前的采样方法是不适合亚临床感染的检测。需要进行非致命性筛查，以防止蛙病毒的进一步传播和风险物种的损失。为了评估非致死性采样方法，我们进行了两个实验：将普通青蛙浴暴露于三种浓度的RUK 13蛙病毒，并将普通蟾蜍暴露于RUK 13或PDE 18。非致死性采样包括暴露后三个时间点采集的颊拭子、手指拭子、身体拭子和水箱拭子，以及沿着脚趾夹和粪便。使用定量PCR在从相同安乐死动物（包括动物）获得的11种不同组织中检查蛙病毒的存在/载量。肝、胃肠道和肾）。颊拭子筛查在两个物种中具有最高的病毒检出率（62%青蛙; 71%蟾蜍），并且与其他非致死性测定相比，产生了持续高的病毒水平。口腔拭子在感染的多个阶段和不同的感染强度下有效，尽管低水平的感染更难检测。口腔拭子检测竞争，甚至优于，青蛙和蟾蜍，分别致死采样。在没有临床症状的情况下观察到成功的病毒检测（33%的青蛙; 50%的蟾蜍）;我们发现RUK 13和PDE 18的可检测性没有差异。我们的研究结果表明，口腔拭子可以取代致命的采样筛选，并作为标准的做法引入蛙病毒监测。
Ranaviruses have been involved in amphibian mass mortality events worldwide. Effective screening to control this pathogen is essential; however, current sampling methods are unsuitable for the detection of subclinical infections. Non-lethal screening is needed to prevent both further spread of ranavirus and losses of at-risk species. To assess non-lethal sampling methods, we conducted two experiments: bath exposing common frogs to RUK13 ranavirus at three concentrations, and exposing common toads to RUK13 or PDE18. Non-lethal sampling included buccal, digit, body and tank swabs, along with toe clips and stool taken across three time-points post-exposure. The presence/load of ranavirus was examined using quantitative PCR in 11 different tissues obtained from the same euthanised animals (incl. liver, gastro-intestinal tract and kidney). Buccal swab screening had the highest virus detection rate in both species (62% frogs; 71% toads) and produced consistently high virus levels compared to other non-lethal assays. The buccal swab was effective across multiple stages of infection and differing infection intensities, though low levels of infection were more difficult to detect. Buccal swab assays competed with, and even outperformed, lethal sampling in frogs and toads, respectively. Successful virus detection in the absence of clinical signs was observed (33% frogs; 50% toads); we found no difference in detectability for RUK13 and PDE18. Our results suggest that buccal swabbing could replace lethal sampling for screening and be introduced as standard practice for ranavirus surveillance.
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