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通过聚合酶链反应分析和生物测定确定商业饲料添加剂作为潜在的猪流行性腹泻病毒缓解策略的影响。

Determining the impact of commercial feed additives as potential porcine epidemic diarrhea virus mitigation strategies as determined by polymerase chain reaction analysis and bioassay.

作者信息

Gebhardt Jordan T, Woodworth Jason C, Jones Cassandra K, Tokach Mike D, Gauger Philip C, Main Rodger G, Zhang Jianqiang, Chen Qi, DeRouchey Joel M, Goodband Robert D, Stark Charles R, Bergstrom Jon R, Bai Jianfa, Dritz Steve S

机构信息

Department of Animal Sciences and Industry, College of Agriculture, Kansas State University, Manhattan, KS.

Department of Veterinary Diagnostic & Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA.

出版信息

Transl Anim Sci. 2018 Aug 20;3(1):93-102. doi: 10.1093/tas/txy100. eCollection 2019 Jan.

Abstract

Mitigation of porcine epidemic diarrhea virus () was assessed using two feed additives (0.5% inclusion of a benzoic acid [] product and 0.02% inclusion of an essential oil [] product; DSM Nutritional Products Inc., Parsippany, NJ), and combination of both products (0.5% BA and 0.02% EO) in spray-dried porcine plasma () and a swine gestation diet (FEED) as determined by real-time quantitative reverse transcriptase polymerase chain reaction () and bioassay. Viral RNA quantification was performed at 7 sampling days post-laboratory inoculation (d 0, 1, 3, 7, 14, 21, and 42) and infectivity was assessed via bioassay with 10-d-old pigs. There was a tendency for treatment × feed matrix × day interaction ( = 0.094), in which the cycle threshold () value increased over time in FEED when treated with both feed additives, whereas there was no increase over time observed in SDPP treated with both feed additives. There was a feed matrix × day interaction ( < 0.001) in which Ct increased over time in FEED, whereas very little increase over time was observed in SDPP. A tendency for a treatment × feed matrix effect ( = 0.085) was observed where FEED treated with the combination of EO and BA had a greater ( < 0.05) PEDV Ct value than other FEED treatments, and all SDPP treatments had the lower PEDV Ct values compared to FEED treatments ( < 0.05). Overall, the combination of both feed additives was most effective at reducing the quantity of genetic material as detected by qRT-PCR ( < 0.001) compared to either additive alone or no feed additive. Virus shedding was observed in the d 7 postinoculation SDPP treatment that was treated with both feed additives, as well as d 0 untreated FEED and d 0 FEED treated with both feed additives. No other treatment bioassay room had detectible RNA shed and detected in fecal swabs or cecal contents. In summary, the combination of EO and BA enhanced the degradation of PEDV RNA in feed but had little impact on RNA degradation in SDPP. Both untreated feed and feed treated with the combination of EO and BA resulted in infection at d 0 post-laboratory inoculation; however, neither set of samples was infective at d 1 postinoculation. In addition, SDPP harbored greater levels of quantifiable RNA for a longer duration of time compared to FEED, and these viral particles remained viable for a longer duration of time indicating differences in viral stability exist between different feed matrices.

摘要

使用两种饲料添加剂(分别添加0.5%的苯甲酸[BA]产品和0.02%的精油[EO]产品;帝斯曼营养产品公司,新泽西州帕西帕尼)以及两种产品的组合(0.5% BA和0.02% EO),通过实时定量逆转录聚合酶链反应(qRT-PCR)和生物测定法,在喷雾干燥猪血浆(SDPP)和母猪妊娠日粮(FEED)中评估对猪流行性腹泻病毒(PEDV)的缓解作用。在实验室接种后的7个采样日(第0、1、3、7、14、21和42天)进行病毒RNA定量,并通过对10日龄仔猪的生物测定法评估感染性。存在处理×饲料基质×天数的交互作用趋势(P = 0.094),即当用两种饲料添加剂处理FEED时,循环阈值(Ct)值随时间增加,而用两种饲料添加剂处理的SDPP中未观察到随时间增加。存在饲料基质×天数的交互作用(P < 0.001),其中FEED中的Ct随时间增加,而SDPP中随时间增加很少。观察到处理×饲料基质效应的趋势(P = 0.085),即用EO和BA组合处理的FEED的PEDV Ct值比其他FEED处理更高(P < 0.05),并且与FEED处理相比,所有SDPP处理的PEDV Ct值更低(P < 0.05)。总体而言,与单独使用任何一种添加剂或不使用饲料添加剂相比,两种饲料添加剂的组合在通过qRT-PCR检测减少遗传物质数量方面最有效(P < 0.001)。在用两种饲料添加剂处理的接种后第7天的SDPP处理中观察到病毒排出,以及未处理的第0天FEED和用两种饲料添加剂处理的第0天FEED中也观察到病毒排出。没有其他处理生物测定室在粪便拭子或盲肠内容物中检测到可检测的RNA排出。总之,EO和BA的组合增强了饲料中PEDV RNA的降解,但对SDPP中的RNA降解影响很小。未处理的饲料和用EO和BA组合处理的饲料在实验室接种后第0天均导致感染;然而,两组样品在接种后第1天均无感染性。此外,与FEED相比,SDPP在更长的时间内含有更高水平的可定量RNA,并且这些病毒颗粒在更长的时间内保持存活,表明不同饲料基质之间存在病毒稳定性差异。

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