UNS: Mechanisms of UV inactivation of viruses for tailored disinfection applications

1512616Rodriguez这项提议的目标是对紫外线对病毒的作用有基本的了解。这项研究将检测分子水平的病毒对紫外线照射的反应，以改进目前的紫外线消毒和未来紫外线技术的设计。这项研究旨在回答有关紫外线灭活病毒的一些基本问题，这些问题可以为工程实践、公共卫生和随后的监管政策提供信息，从而产生广泛的影响。拟议的活动将促进一些关键领域的知识。在消毒实践中，新的基础知识将发展到病毒对紫外线照射的分子水平的反应中。具体地说，除了腺病毒，还将对另一种双链DNA病毒--JC多瘤病毒--进行新的研究，以测试腺病毒对低压和中压紫外线反应的研究结果的普适性。这项研究将超越对腺病毒实验室毒株的传统评估，分离和测试环境毒株，以确定紫外线反应是否存在任何差异，这可能与重复暴露和在实验室细胞培养株中繁殖的驯化反应(如DNA修复)有关。将在分子微生物学方法的一些新领域进行开发，其中一些是我们开发的，包括特定紫外光波长对早期感染的抑制，跨病变合成的DNA修复，使用电泳和质谱学技术评估蛋白质损伤，以及使用我们最近发表的一种新的远程PCR技术进行DNA损伤。这些基本发现将被用来开发一种基于定制波长紫外线-LED的节能辐射系统，以比传统紫外线源更低的成本优化消毒。本文提出的工作将：(1)评估两种双链DNA病毒的失活动力学和波长敏感性，(2)将腺病毒研究扩大到从环境中分离的腺病毒，(3)评估DNA修复在细胞培养感染性结果中的基本作用，(4)采用新开发的基于分子生物学的分析方法来评估不同波长的紫外光照射对DNA损伤和蛋白质损伤的影响，以及(5)了解由于纳米专用紫外光发光二极管(LED)的开发而可能采用的定制波长方法对消毒的影响。监管机构和公共卫生官员需要信息来帮助公共部门做出明智的决策。小型系统的病毒消毒问题目前存在争议，因为地下水规则明确禁止使用紫外线对任何病毒灭活积分，因为腺病毒灭活所需的高剂量目前无法在紫外线反应器中进行现场验证。因此，如果紫外线消毒对地下水中的病毒消毒起到任何作用，这项研究的时机是至关重要的。因此，希望使用紫外线保护公共健康的小系统可能会受到不必要的影响。

1512616RodriguezThe goal of this proposal is to gain fundamental insights into the action of ultraviolet light on viruses. The study will examine the molecular level viral responses to ultraviolet irradiation to improve current ultraviolet light disinfection and design of future ultraviolet light technologies. The research aims to answer a number of fundamental questions around virus inactivation with ultraviolet light that can inform engineering practice, public health, and subsequent regulatory policy and thus has broad reaching impacts.The proposed activity will advance knowledge in a number of key areas. New fundamental knowledge will be developed into the molecular-level response of viruses to ultraviolet light irradiation during disinfection practice. Specifically, in addition to adenoviruses, new studies will be performed on another double stranded DNA virus - JC polyomavirus - to test the generalizability of the findings with adenoviruses in response to low-pressure and medium-pressure ultraviolet light. This research will move beyond the traditional evaluations of laboratory strains of adenoviruses and isolate and test environmental strains to determine if there are any differences in the ultraviolet light -response that may be related to acclimated responses (such as DNA repair) from repeated exposure to and propagation in laboratory cell culture lines. A number of new areas in molecular microbiology methods, some of which we have developed, will be exploited including early stage infection inhibition due to specific ultraviolet light wavelengths, DNA repair from trans-lesion synthesis, protein damage assessment using electrophoresis and mass spectrometry techniques, and DNA damage using a new long range PCR technique we recently published. These fundamental findings will be used to develop a tailored wavelength ultraviolet-LED based energy efficient irradiation system to optimize disinfection at lower cost than conventional ultraviolet sources. The work proposed herein will: (1) evaluate inactivation kinetics and wavelength sensitivity of two double stranded DNA viruses, (2) extend the adenovirus studies to adenoviruses isolated from the environment, (3) evaluate the fundamental role of DNA repair in the cell culture infectivity outcomes, (4) incorporate newly developed molecular biology-based assays to assess DNA damage and protein damage from ultraviolet light irradiation at varying wavelengths, and, (5) understand the implications for disinfection using tailored wavelength approaches made possible by the development of nanometer-specific ultraviolet light emitting diodes (LEDs). Regulators and public health officials need information to help make sound decisions for the public sector. The issue of virus disinfection for small systems is currently contentious as the Groundwater Rule specifically forbids the use of UV for any virus inactivation credit due to the high doses required for adenovirus inactivation that cannot currently be field-validated in UV reactors. Therefore, the timing of this research is critical if UV disinfection is going to play any role for virus disinfection in groundwater. Thus, small-systems wanting to use UV for protection of public health, may needlessly suffer.