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经小鼠适应的羊瘙痒病的碱性水解,用于朊病毒阳性材料的灭活与处置。

Alkaline hydrolysis of mouse-adapted scrapie for inactivation and disposal of prion-positive material.

作者信息

Murphy R G L, Scanga J A, Powers B E, Pilon J L, Vercauteren K C, Nash P B, Smith G C, Belk K E

机构信息

Center for Meat Safety and Quality, Department of Animal Sciences, Colorado State University, Fort Collins, CO 80523, USA.

出版信息

J Anim Sci. 2009 May;87(5):1787-93. doi: 10.2527/jas.2008-1492. Epub 2008 Dec 19.

Abstract

Prion diseases such as bovine spongiform encephalopathy, chronic wasting disease, and scrapie pose serious risks to human and animal health due to a host of disease-specific factors, including the resistance of infectious prions (PrP(Sc)) to natural degradation and to most commercial inactivation procedures. In an attempt to address this concern, a mouse model was used to compare the efficacy of an alkaline hydrolysis process with a simulated continuous-flow rendering treatment for disposal of PrP(Sc)-infected biological material. Female C57/BL6 mice (N = 120) were randomly divided into 4 treatment groups (n = 30), and each mouse was injected intraperitoneally with their designated treatment inoculum. Treatment groups 1 and 2 served as the positive and negative controls, respectively. Group 3 was inoculated with rendered scrapie-positive mouse brain material to investigate the effectiveness of simulated continuous-flow rendering practices to reduce or eliminate PrP(Sc). Group 4 was inoculated with hydrolyzed scrapie-positive mouse brain material to determine the sterilizing effect of alkaline hydrolysis on PrP(Sc). Mice were monitored for overt signs of disease, and those showing clinical signs were killed to prevent undue suffering. Brains were obtained from all mice that died (or were killed) and analyzed with an ELISA for the presence of PrP(Sc). Results indicated that the simulated continuous-flow rendering treatment used for preparing the rendering treatment group inoculum failed to completely eliminate PrP(Sc). Rendering delayed, but did not stop, clinical mouse-adapted scrapie transmission. Compared with positive controls, the rendering treatment group experienced an approximate 45-d average delay in days to death (250 vs. 205 d for positive controls; P < 0.0001) and a death loss of 73.9% (P = 0.0094). Positive controls suffered 100% death loss. The results validated the efficacy of the alkaline hydrolysis treatment to inactivate all PrP(Sc) because no alkaline hydrolysis treatment group mice succumbed to the disease (P < 0.0001). Based on our results, alkaline hydrolysis should be considered by the animal rendering and beef packing industries as an alternative to incineration, landfill burial, and rendering for disposing of biological material potentially infected or contaminated with prion disease.

摘要

诸如牛海绵状脑病、慢性消耗病和羊瘙痒症等朊病毒疾病,由于一系列特定于疾病的因素,对人类和动物健康构成严重风险,这些因素包括传染性朊病毒(PrP(Sc))对自然降解以及大多数商业灭活程序的抗性。为了解决这一问题,使用了一种小鼠模型来比较碱性水解过程与模拟连续流炼制处理对处理感染PrP(Sc)的生物材料的效果。将雌性C57/BL6小鼠(N = 120)随机分为4个处理组(n = 30),每只小鼠腹腔注射指定的处理接种物。处理组1和2分别作为阳性和阴性对照。第3组接种经过炼制的羊瘙痒症阳性小鼠脑材料,以研究模拟连续流炼制操作减少或消除PrP(Sc)的有效性。第4组接种经水解的羊瘙痒症阳性小鼠脑材料,以确定碱性水解对PrP(Sc)的杀菌效果。监测小鼠的明显疾病迹象,对出现临床症状的小鼠实施安乐死以防止过度痛苦。从所有死亡(或被安乐死)的小鼠获取大脑,并用ELISA分析PrP(Sc)的存在情况。结果表明,用于制备炼制处理组接种物的模拟连续流炼制处理未能完全消除PrP(Sc)。炼制延迟了,但并未阻止临床适应小鼠的羊瘙痒症传播。与阳性对照相比,炼制处理组的平均死亡天数延迟约45天(阳性对照为205天,炼制处理组为250天;P < 0.0001),死亡损失率为73.9%(P = 0.0094)。阳性对照的死亡损失率为100%。结果证实了碱性水解处理灭活所有PrP(Sc)的有效性,因为没有碱性水解处理组的小鼠死于该疾病(P < 0.0001)。基于我们的结果,动物炼制和牛肉包装行业应考虑将碱性水解作为焚烧、填埋和炼制的替代方法,用于处理可能感染或被朊病毒疾病污染的生物材料。

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