基于模式病毒评价商品化消毒剂对非洲猪瘟病毒的消杀效果

doi:10.16178/j.issn.0528-9017.20230621

浙江农业科学 ›› 2024, Vol. 65 ›› Issue (8): 1944-1949.DOI: 10.16178/j.issn.0528-9017.20230621

基于模式病毒评价商品化消毒剂对非洲猪瘟病毒的消杀效果

倪征¹(), 朱寅初¹, 孙冰冰², 云涛¹, 陈柳¹, 徐辉², 叶伟成¹, 华炯钢¹, 张存¹^,^*()

1.浙江省农业科学院畜牧兽医研究所,浙江杭州 310021
2.浙江省动物疫病预防控制中心,浙江杭州 310018

收稿日期:2023-12-26 出版日期:2024-08-11 发布日期:2024-08-21
通讯作者: 张存(1965-),男,研究员,主要从事动物病毒学研究,E-mail:zhangcun@aliyun.com。
作者简介:倪征(1979-),女,浙江杭州人,助理研究员,硕士,主要从事动物病毒学研究,E-mail:nizheng79@163.com。
基金资助:
浙江省“三农六方”科技协作项目(2019SNLF018);浙江省重点研发计划(2021C02050-2)

Evaluation of inactivation effect of commercial disinfectant on African swine fever virus based on model virus

NI Zheng¹(), ZHU Yinchu¹, SUN Bingbing², YUN Tao¹, CHEN Liu¹, XU Hui², YE Weicheng¹, HUA Jionggang¹, ZHANG Cun¹^,^*()

1. Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang
2. Zhejiang Provincial Center for Animal Disease Control, Hangzhou 310018, Zhejiang

Received:2023-12-26 Online:2024-08-11 Published:2024-08-21

摘要/Abstract

摘要：

为了解常用商品化消毒剂对非洲猪瘟病毒(ASFV)的消杀效果,为其消毒剂的筛选提供参考。该研究利用伪狂犬病毒(pseudorabies virus,PRV)与ASFV病毒理化特征的相似性,以PRV作为替代标识毒株,建立了消毒剂细胞评价模型,于体外检测了3种市售消毒剂对生长在BHK-21细胞中PRV的灭活效果。分别从3类消毒剂的不同方面对消毒剂抗病毒作用的影响进行消杀效果的评估。研究显示,有效含量为0.125 g·L^-1的戊二醛癸甲溴铵溶液,以及有效含量为0.5 g·L^-1的过氧乙酸溶液和二氯异氰脲酸钠,在室温作用30 min,均可有效灭活PRV。其中,过氧乙酸综合评价最优;戊二醛癸甲溴铵性价比最高,但所需消杀时间较长,受低温影响比较严重。该研究可为养殖场防控ASFV的感染提供消毒剂筛选的理论依据,并对ASFV的消毒标准提供参考。

关键词: 消毒剂细胞评价模型, ASFV, PRV, 杀灭病毒效果

Abstract:

In order to evaluate and prove the elimination effect of commercial disinfectants on African swine fever virus (ASFV), and to provide reference for the screening of disinfectants. In this study, the similarity of physicochemical characteristics of pseudorabies virus (PRV) and ASFV virus was used to establish a disinfectant cell evaluation model with PRVas an alternative marker strain. The inactivation effect of three commercially available disinfectants on PRV grown in HBK-21 cells was tested in vitro. The disinfecting effect of the three disinfectants was evaluated from several aspects on the antiviral effect of the three disinfectants. The results showed that glutaraldehyde methyl bromide solution with an effective content of 0.125 g·L^-1, peracetic acid solution with an effective content of 0.5 g·L^-1 and sodium dichloroisocyanurate could effectively inactivate PRV at room temperature for 30 min. Among them, peracetic acid comprehensive evaluation is the best; Glutaraldehyde decyl methyl bromide is the most cost-effective, but it requires a long time of elimination and is seriously affected by low temperature. This study can provide theoretical basis for disinfectant screening for the prevention and control of ASFV infection in livestock farms, and provide reference for the establishment of disinfection standards for ASFV.

Key words: disinfectant cell evaluation model, African swine fever virus(ASFV), pseudorabies virus(PRV), virus killing effect

中图分类号:

S852

倪征, 朱寅初, 孙冰冰, 云涛, 陈柳, 徐辉, 叶伟成, 华炯钢, 张存. 基于模式病毒评价商品化消毒剂对非洲猪瘟病毒的消杀效果[J]. 浙江农业科学, 2024, 65(8): 1944-1949.

NI Zheng, ZHU Yinchu, SUN Bingbing, YUN Tao, CHEN Liu, XU Hui, YE Weicheng, HUA Jionggang, ZHANG Cun. Evaluation of inactivation effect of commercial disinfectant on African swine fever virus based on model virus[J]. Journal of Zhejiang Agricultural Sciences, 2024, 65(8): 1944-1949.

图/表 7

表1 本试验所用消毒剂名称和主要成分

Table 1 The name and main ingredients of the disinfectant used in this experiment

序号	消毒剂名称	主要成分	物理性状	批准文号
1	戊二醛癸甲溴铵	5%戊二醛+5%癸甲溴铵	溶液	兽药字230036245
2	过氧乙酸	20%过氧乙酸	溶液	兽药字180206432
3	二氯异氰脲酸钠	20%有效氯	白色粉末	兽药字180086051

图1 PRV在BHK-21上的增殖 A-正常BHK-21细胞对照;B-病毒感染细胞24 h;C-病毒感染细胞48 h;D-病毒感染细胞72 h。

Fig.1 The proliferation of PRV on BHK-21

表2 病毒TCID50的测定

Table 2 Determination of TCID50

病毒稀释倍数	有CPE孔	无CPE孔	累计 CPE数量	累计无 CPE数量	合计	感染百分率/%
10^-1	8	0	55	0	55/55	100
10^-2	8	0	47	0	47/47	100
10^-3	8	0	39	0	39/39	100
10^-4	8	0	31	0	31/31	100
10^-5	8	0	23	0	23/23	100
10^-6	8	0	15	0	15/15	100
10^-7	5	3	7	3	7/10	70
10^-8	2	6	2	9	2/11	18
10^-9	0	8	0	17	0/17	0

图2 消毒剂对病毒增殖的影响 A-细胞接种病毒后的特异性荧光(200×);B-病毒与消毒剂作用的实验组1(200×);C-病毒与消毒剂作用的实验组2(200×);D-病毒完全消杀,细胞无绿色荧光(200×)。

Fig.2 Effect of disinfectants on virus proliferation

表3 3种消毒剂在不同稀释倍数下的病毒灭活率分析

Table 3 Analysis of virus inactivation rate of three disinfectants at different dilution times

稀释倍数	病毒灭活率/%
稀释倍数	戊二醛癸甲溴铵	过氧乙酸	二氯异氰脲酸钠
1∶100	-	100	100
1∶200	-	100	100
1∶400	100	100	100
1∶800	100	68	32
1∶1 600	21	35	8

图3 3种消毒剂最短起效时间及不同作用时间对消毒效果的影响

Fig.3 Analysis of the shortest onset time of three disinfectants and the influence of different action times on the disinfection effect

图4 温度对消毒剂消毒效果影响的结果 a-戊二醛癸甲溴铵溶液的灭活效率;b-过氧乙酸溶液的灭活效率;c-二氯异氰脲酸钠的灭活效率。

Fig.4 The result of the influence of temperature on the disinfection effect of disinfectants

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基于模式病毒评价商品化消毒剂对非洲猪瘟病毒的消杀效果

Evaluation of inactivation effect of commercial disinfectant on African swine fever virus based on model virus

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