Wastewater-based epidemiology (WBE) has proven to be a powerful tool for tracking the spread of viral pathogens, such as SARS-CoV-2; however, as WBE has expanded to include new pathogens, like mpox virus (MPXV), more data is needed to guide practitioners on how to design WBE campaigns. Here, we investigated the decay rates of heat-inactivated MPXV (HI-MPXV) and attenuated vaccinia virus (VV) viral signal in primary influent and settled solids collected from a local POTW at 4°C, 22°C, or 35°C using digital PCR. Subsequently, we studied the solid-liquid partitioning of the viruses in primary influent. Over 30 days, decay rates did not significantly differ between viruses (p=0.5258). However, decay was significantly higher in primary influent (0.109-0.144/day) than in settled solids (0.019-0.040/day) at both 22°C (p=0.0030) and 35°C (p=0.0166). Furthermore, as part of the partitioning experiment, we found that HI-MPXV and VV adsorb to the solids fraction of primary influent at higher intensities than previously studied enveloped viruses (K_F=1,000-31,800 mL/g, n=1.01-1.41). Likewise, it was determined in the partitioning experiment that a concentration of greater than 10³ gc/mL in raw influent was needed for the viable quantification of MPXV and VV DNA in the clarified liquid fraction of raw primarily influent. Our study provides essential insights into informative sample collection and storage conditions for the analysis of wastewater and for transport modeling studies. Due to the slow decay observed in settled solids at all tested temperatures in the persistence experiment, this matrix may be most suitable for retrospective analyses of MPXV infections within a community.

中文翻译：

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原代进水和沉降固体中 mpox 和痘苗病毒 DNA 的衰变和固液分配指导基于废水的流行病学实践

基于废水的流行病学 （WBE） 已被证明是追踪病毒病原体传播的有力工具，例如 SARS-CoV-2;然而，随着 WBE 已经扩展到包括新的病原体，如 mpox 病毒 （MPXV），需要更多数据来指导从业者如何设计 WBE 活动。在这里，我们使用数字 PCR 研究了在 4°C、22°C 或 35°C 下从本地 POTW 收集的初级进水和沉降固体中热灭活 MPXV （HI-MPXV） 和减毒痘苗病毒 （VV） 病毒信号的衰减率。随后，我们研究了病毒在原源进水中固液的分配。在 30 天内，病毒之间的腐烂率没有显著差异 （p=0.5258）。然而，在 22°C （p=0.0030） 和 35°C （p=0.0166） 下，初级进水的腐烂 （0.109-0.144/天） 显著高于沉降固体 （0.019-0.040/天）。此外，作为分配实验的一部分，我们发现 HI-MPXV 和 VV 以比先前研究的包膜病毒更高的强度吸附到初级进水的固体部分 （K_F=1,000-31,800 mL/g，n=1.01-1.41）。同样，在分配实验中确定，需要原始进水中大于 10³ gc/mL 的浓度才能对原始主要进水的澄清液体部分中的 MPXV 和 VV DNA 进行有效定量。我们的研究为废水分析和运输建模研究提供了信息丰富的样品采集和储存条件的重要见解。由于在持久性实验中在所有测试温度下观察到的沉降固体缓慢衰变，因此该基质可能最适合对社区内的 MPXV 感染进行回顾性分析。