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堆肥作为猪流行性腹泻病毒(PEDV)感染猪尸体生物安全处置方法的有效性。

Effectiveness of composting as a biosecure disposal method for porcine epidemic diarrhea virus (PEDV)-infected pig carcasses.

作者信息

Vitosh-Sillman Sarah, Loy John Dustin, Brodersen Bruce, Kelling Clayton, Eskridge Kent, Millmier Schmidt Amy

机构信息

School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Fair Street and East Campus Loop, Lincoln, NE 68583 USA.

Department of Statistics, University of Nebraska-Lincoln, Lincoln, NE 68583 USA.

出版信息

Porcine Health Manag. 2017 Nov 28;3:22. doi: 10.1186/s40813-017-0068-z. eCollection 2017.

Abstract

BACKGROUND

Porcine epidemic diarrhea virus (PEDV) is an enteric disease of swine that has emerged as a worldwide threat to swine herd health and production. Substantial research has been conducted to assess viability of the virus on surfaces of vehicles and equipment, in feed and water, and on production building surfaces, but little is known about the persistence in PEDV-infected carcasses and effective disposal methods thereof. This study was conducted to quantify the persistence of PEDV RNA via quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) at various time-temperature combinations and in infected piglet carcasses subjected to composting. Although this method does not distinguish between infectious and noninfectious virus, it is a rapid and sensitive test to evaluate materials for evidence of virus genome.

RESULTS

In the first study, PEDV was suspended in cell culture media at 1 × 10 TCID50 per sample (1 mL sample size) and subjected to various time and temperature combinations in triplicate including temperatures of 37, 45, 50, 55, 60, 65, 70 °C and exposure times of 0, 1, 2, 3, 4, 5, 7, and 14 days. At all temperatures, viral RNA copies declined over time, with the decline most marked and rapid at 65 and 70 °C. Detectable RNA did persist throughout the trial in all but the most extreme condition, where two of three samples incubated at 70 °C yielded undetectable viral RNA after 14 days. In the second study, PEDV-infected piglet carcasses were subjected to two cycles of composting lasting 36 and 37 days, respectively, for a total compost time of 73 days. Composting was performed in triplicate windrow sections housed inside biosecure, climate-controlled rooms using insulated bins designed to represent a continuous windrow compost pile. Temperatures reached 35-57 °C for 26 days of cycle 1 and 35-45 °C for 3 days of cycle 2. Samples consisting of carbon material with or without decomposed tissue as available per sample site collected at ten locations throughout the cross-section of each windrow section following the primary and secondary compost cycles yielded no detectable viral RNA.

CONCLUSIONS

Composting appears to be an effective disposal method for PEDV-infected piglet carcasses under the conditions examined. The combination of time and high temperature of the compost cycle effectively degraded viral RNA in cell culture media that should provide optimum stability. Complex compost material matrices collected from windrow sections yielded undetectable PEDV RNA by qRT-PCR after one 36-day compost cycle despite incomplete decomposition of soft tissue.

摘要

背景

猪流行性腹泻病毒(PEDV)是一种猪肠道疾病,已成为对猪群健康和生产的全球性威胁。已开展了大量研究来评估该病毒在车辆和设备表面、饲料和水中以及生产建筑表面的存活能力,但对于PEDV感染尸体中的病毒持久性及其有效处置方法知之甚少。本研究旨在通过定量实时逆转录聚合酶链反应(qRT-PCR)在不同时间 - 温度组合下以及在进行堆肥处理的感染仔猪尸体中定量检测PEDV RNA的持久性。尽管该方法无法区分感染性和非感染性病毒,但它是一种快速且灵敏的测试方法,可用于评估材料中病毒基因组的证据。

结果

在第一项研究中,将PEDV以每个样本1×10 TCID50(样本体积1 mL)悬浮于细胞培养基中,并一式三份进行各种时间和温度组合处理,温度包括37、45、50、55、60、65、70°C,暴露时间为0、1、2、3、4、5、7和14天。在所有温度下,病毒RNA拷贝数随时间下降,在65和70°C时下降最为明显和迅速。在除最极端条件外的所有试验中,均可检测到RNA持续存在,在70°C孵育的三个样本中有两个在14天后未检测到病毒RNA。在第二项研究中,对感染PEDV的仔猪尸体分别进行了两个持续36天和37天的堆肥周期,总堆肥时间为73天。堆肥在生物安全、气候控制的房间内的三个平行条垛中进行,使用设计成代表连续条垛式堆肥堆的隔热箱。第一个周期26天温度达到35 - 57°C,第二个周期3天温度达到35 - 45°C。在每个条垛的横断面的十个位置,在初级和次级堆肥周期后收集的样本(由每个样本位点的含或不含分解组织的碳材料组成)均未检测到病毒RNA。

结论

在所研究的条件下,堆肥似乎是处理感染PEDV的仔猪尸体的有效方法。堆肥周期的时间和高温组合有效地降解了细胞培养基中的病毒RNA,而细胞培养基应能提供最佳稳定性。尽管软组织未完全分解,但在一个36天的堆肥周期后,从条垛中收集的复杂堆肥材料基质经qRT-PCR检测未检测到PEDV RNA。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/472b/5704383/a382de7554ea/40813_2017_68_Fig1_HTML.jpg

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