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Article(id=1241771802217743180, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241771801366299468, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202312247, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1702569600000, receivedDateStr=2023-12-15, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773991068652, onlineDateStr=2026-03-20, pubDate=1729785600000, pubDateStr=2024-10-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773991068652, onlineIssueDateStr=2026-03-20, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773991068652, creator=13701087609, updateTime=1773991068652, updator=13701087609, issue=Issue{id=1241771801366299468, tenantId=1146029695717560320, journalId=1227665162245664772, year='2024', volume='51', issue='20', pageStart='3649', pageEnd='3840', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773991068450, creator=13701087609, updateTime=1773991676896, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241774353457681403, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241771801366299468, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241774353457681404, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1241771801366299468, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=3691, endPage=3694, ext={EN=ArticleExt(id=1241771802528121679, articleId=1241771802217743180, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Surveillance and analysis of enteroviruses and diarrheal viruses in environmental sewage, Aba, 2022, columnId=null, journalTitle=Modern Preventive Medicine, columnName=null, runingTitle=null, highlight=null, articleAbstract=
Objective

To assess the presence of enteroviruses and diarrheal viruses in the environmental sewage of Aba in 2022, and to provide evidence supporting effective control measures against these viruses.

Methods

Sewage samples were collected monthly throughout 2022, then eluted and concentrated by anionic membrane adsorption.Through this, enteroviruses were isolated and identified. At the same time, nucleic acid was extracted from the concentrated liquid, and real-time fluorescent PCR was used to detect several diarrhea viruses, such as group A rotavirus, norovirus, Sapovirus, human astrovirus and enteric adenovirus.

Results

A total of 12 environmental sewage samples were collected from January to December 2022. Out of these, 3 strains of polioviruses and 9 non-polio enteroviruses were isolated. All 3 poliovirus strains were detected as Type 3 vaccine-like poliovirus (PV) strains. Group A rotavirus and norovirus genotype II were detected every month. Norovirus type I and enteric adenovirus were observed in all months except November. Human astrovirus was absent in samples from July, August, November and December. Sapovirus was not identified throughout the year.

Conclusion

Enteroviruses and diarrheal viruses exist widely in Aba’s environmental sewage. Environmental sewage surveillance can offer a comprehensive view of these viruses’ etiological and epidemiological characteristics. Continuous environmental sewage surveillance can predict and warn the epidemic and outbreak of enteroviruses and diarrheal virus-related diseases.

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目的

了解2022年阿坝州环境污水中的肠道病毒和腹泻病毒的检出情况，为有效控制肠道病毒和腹泻病毒的流行提供依据。

方法

2022年每月在阿坝州采集环境污水，通过阴离子膜吸附法洗脱浓缩后，进行肠道病毒分离和鉴定；同时浓缩液提取核酸，通过实时荧光定量PCR测定诺如病毒、A组轮状病毒、札如病毒、肠道腺病毒、人星状病毒几种腹泻病毒。

结果

2022年1—12月共采集12份环境污水，共分离出脊髓灰质炎病毒3株和非脊灰肠道病毒9株，3株脊髓灰质炎病毒均为Ⅲ型脊灰疫苗相似株。A组轮状病毒和诺如病毒Ⅱ型每月均有检出；诺如病毒Ⅰ型和肠道腺病毒除11月外其他月份均有检出；人星状病毒在7月、8月、11月、12月未能检出；札如病毒全年未检出。

结论

阿坝州环境污水中肠道病毒和腹泻病毒广泛存在，环境污水监测能够全面了解肠道病毒和腹泻病毒病原学特征和流行特征，持续的环境污水监测可为肠道病毒和腹泻病毒相关疾病的流行和暴发做出预测预警。

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马小珍，E-mail：
, copyrightStatement=本刊刊出的所有文章不代表中华预防医学会和本刊编委会的观点，除非特别声明。, copyrightOwner=中华预防医学会和四川大学华西公共卫生学院, extLink=null, articleAbsUrl=null, sourceXml=gl+7V33d28XABPy8bS86jA==, magXml=yqnLLlAM5Rz5XNecjrLgLw==, pdfUrl=null, pdf=M2O5/DVnyaBiqcin2ECOow==, pdfFileSize=468616, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=bE/cLa1KRaZP1fjRmtFCSg==, mapNumber=null, authorCompany=null, fund=null, authors=

张章（1973—），男，大专，副主任技师，病原微生物检验研究

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张章（1973—），男，大专，副主任技师，病原微生物检验研究

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张章（1973—），男，大专，副主任技师，病原微生物检验研究

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Results of enterovirus and diarrheal virus in environmental sewage in Aba 2022

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月份肠道病毒腹泻病毒
肠道病毒分离结果脊灰病毒分离株数非脊灰肠道病毒分离株数A组轮状病毒诺如病毒GⅠ型诺如病毒GⅡ型札如病毒人星状病毒肠道腺病毒
1月-//+++-++
2月+02+++-++
3月+03+++-++
4月+22+++-++
5月+11+++-++
6月+01+++-++
7月-//+++--+
8月-//+++--+
9月-//+++-++
10月-//+++-++
11月-//+-+---
12月-//+++--+
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2022年阿坝州环境污水中肠道病毒与腹泻病毒的检测结果

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月份肠道病毒腹泻病毒
肠道病毒分离结果脊灰病毒分离株数非脊灰肠道病毒分离株数A组轮状病毒诺如病毒GⅠ型诺如病毒GⅡ型札如病毒人星状病毒肠道腺病毒
1月-//+++-++
2月+02+++-++
3月+03+++-++
4月+22+++-++
5月+11+++-++
6月+01+++-++
7月-//+++--+
8月-//+++--+
9月-//+++-++
10月-//+++-++
11月-//+-+---
12月-//+++--+
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2022年阿坝州环境污水肠道病毒和腹泻病毒监测分析
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张章 1 , 陈娜 2 , 曹冉冉 2 , 彭刚 1 , 李雪 1 , 马小珍 2
现代预防医学 | 环境与职业卫生 2024,51(20): 3691-3694
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现代预防医学 | 环境与职业卫生 2024, 51(20): 3691-3694
2022年阿坝州环境污水肠道病毒和腹泻病毒监测分析
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张章1, 陈娜2, 曹冉冉2, 彭刚1, 李雪1, 马小珍2
作者信息
  • 1.阿坝州疾病预防控制中心，四川 马尔康 624050
  • 2.四川省疾病预防控制中心

    • 张章（1973—），男，大专，副主任技师，病原微生物检验研究

通讯作者:

马小珍，E-mail：
Surveillance and analysis of enteroviruses and diarrheal viruses in environmental sewage, Aba, 2022
Zhang ZHANG1, Na CHEN2, Ran-ran CAO2, Gang PENG1, Xue LI1, Xiao-zhen MA2
Affiliations
  • Aba Center for Disease Control and Prevention, Maerkang, Sichuan 624050, China
出版时间: 2024-10-25 doi: 10.20043/j.cnki.MPM.202312247
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摘要
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目的

了解2022年阿坝州环境污水中的肠道病毒和腹泻病毒的检出情况，为有效控制肠道病毒和腹泻病毒的流行提供依据。

方法

2022年每月在阿坝州采集环境污水，通过阴离子膜吸附法洗脱浓缩后，进行肠道病毒分离和鉴定；同时浓缩液提取核酸，通过实时荧光定量PCR测定诺如病毒、A组轮状病毒、札如病毒、肠道腺病毒、人星状病毒几种腹泻病毒。

结果

2022年1—12月共采集12份环境污水，共分离出脊髓灰质炎病毒3株和非脊灰肠道病毒9株，3株脊髓灰质炎病毒均为Ⅲ型脊灰疫苗相似株。A组轮状病毒和诺如病毒Ⅱ型每月均有检出；诺如病毒Ⅰ型和肠道腺病毒除11月外其他月份均有检出；人星状病毒在7月、8月、11月、12月未能检出；札如病毒全年未检出。

结论

阿坝州环境污水中肠道病毒和腹泻病毒广泛存在，环境污水监测能够全面了解肠道病毒和腹泻病毒病原学特征和流行特征，持续的环境污水监测可为肠道病毒和腹泻病毒相关疾病的流行和暴发做出预测预警。

环境污水  /  肠道病毒  /  腹泻病毒  /  阿坝州
Objective

To assess the presence of enteroviruses and diarrheal viruses in the environmental sewage of Aba in 2022, and to provide evidence supporting effective control measures against these viruses.

Methods

Sewage samples were collected monthly throughout 2022, then eluted and concentrated by anionic membrane adsorption.Through this, enteroviruses were isolated and identified. At the same time, nucleic acid was extracted from the concentrated liquid, and real-time fluorescent PCR was used to detect several diarrhea viruses, such as group A rotavirus, norovirus, Sapovirus, human astrovirus and enteric adenovirus.

Results

A total of 12 environmental sewage samples were collected from January to December 2022. Out of these, 3 strains of polioviruses and 9 non-polio enteroviruses were isolated. All 3 poliovirus strains were detected as Type 3 vaccine-like poliovirus (PV) strains. Group A rotavirus and norovirus genotype II were detected every month. Norovirus type I and enteric adenovirus were observed in all months except November. Human astrovirus was absent in samples from July, August, November and December. Sapovirus was not identified throughout the year.

Conclusion

Enteroviruses and diarrheal viruses exist widely in Aba’s environmental sewage. Environmental sewage surveillance can offer a comprehensive view of these viruses’ etiological and epidemiological characteristics. Continuous environmental sewage surveillance can predict and warn the epidemic and outbreak of enteroviruses and diarrheal virus-related diseases.

Environmental sewage  /  Enteroviruses  /  Diarrheal viruses  /  Aba
张章, 陈娜, 曹冉冉, 彭刚, 李雪, 马小珍. 2022年阿坝州环境污水肠道病毒和腹泻病毒监测分析. 现代预防医学, 2024 , 51 (20) : 3691 -3694 . DOI: 10.20043/j.cnki.MPM.202312247
Zhang ZHANG, Na CHEN, Ran-ran CAO, Gang PENG, Xue LI, Xiao-zhen MA. Surveillance and analysis of enteroviruses and diarrheal viruses in environmental sewage, Aba, 2022[J]. Modern Preventive Medicine, 2024 , 51 (20) : 3691 -3694 . DOI: 10.20043/j.cnki.MPM.202312247
正文
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肠道病毒与腹泻性病毒的传染性强，常引起儿童发病，容易在托幼机构和学校中形成聚集感染，给公众健康带来巨大威胁[1]。2018以来，阿坝州肠道传染病的发病率总体呈上升趋势，肠道传染病聚集性疫情多发，对人民群众的健康造成重大风险。
肠道病毒（Enterovirus, EV）属于小核糖核酸病毒科，包含柯萨奇病毒（coxsackievirus, CA）、脊髓灰质炎病毒（Poliovirus, PV）、鼻病毒（rhinovirus, RV）、埃可病毒（echovirus, ECHO）和人肠道病毒（Enteroviruses, EV）A组-D组等非脊灰肠道病毒（Non-Polio Enterovirus）。这些病毒可引起从普通感冒、腹泻、皮疹、到无菌性脑膜炎和脊髓灰质炎等多种疾病[2]。腹泻病毒包含肠道腺病毒（Enteric Adenovirus, EAdV）、A组轮状病毒（Group A Rotavirus, RVA）、札如病毒（Sapovirus, SV）、诺如病毒（Norovirus, NoV）、人星状病毒（Human Astrivirus, HAstV）等，它们主要通过粪-口途径传播。病毒颗粒可随着被感染个体的粪便大量排出进入环境[3-5]。多种肠道病毒和腹泻病毒可在水环境中持久存在，可长时间保持传染性[6]。
环境污水中肠道和腹泻病毒监测，对预测相关疾病的传播致关重要。病毒的环境污水监测是对环境污水中的样本进行富集浓缩后，采用病毒分离、分子生物学技术等方法对目的病毒进行检测的一种监测方法。环境污水监测首先应用于肠道病毒的分子流行病学监测，作为急性弛缓性麻痹（Acute Flaccid Paralysis, AFP）病例监测的一个重要补充[7-8]，在全球消灭脊髓灰质炎行动（Global Polio Eradication Initiative, GPEI）中发挥重要作用[9]。A组轮状病毒、诺如病毒、札如病毒、人星状病毒、肠道腺病毒等几种腹泻病毒与肠道病毒同样都以粪-口途径为主要传播方式[10]，因此环境污水监测也广泛应用于腹泻病毒的监测[4-5,11-12]。
本研究通过对2022年阿坝州环境污水中的多种肠道病毒和腹泻病毒进行监测和分析，旨在了解阿坝州的主要肠道病毒和腹泻病毒病原谱、分析其病原学特征和流行特征、以便及时做出疾病预警。
1　材料与方法
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1.1　环境样本采集
本研究监测地点为阿坝州马尔康市的某个污水处理厂。依照《2023四川省污水监测工作指南》规定的月度监测要求，采样人员于2022年1月—12月分别在该污水处理厂的污水进水口采集水样2L，每月1份，全年共12份。采集样本立即置于4℃保存，并将样本于48小时内低温运输至四川省疾病预防控制中心微生物检验所实验室。如遇特殊情况未能及时运输，采集的标本置于-20 ℃保存并择日低温运输。
1.2　检测方法
1.2.1　环境样本的过滤与洗脱
环境样本的过滤和洗脱按照中国疾病预防控制中心-病毒病预防控制所-国家脊髓灰质炎实验室《病毒环境监测技术培训班操作手册》2018年第6版的操作规程进行。实验前需对装置及剪刀、镊子进行高压灭菌。取1 L污水样品，倒入250 mL离心管中进行离心（4℃，3 000 rpm，30 min）。采集上清，加入MgCl2（2.5 M）使终浓度为0.05 M，进一步加入HCl（0.5 N）将pH值调节至3.5。将硝酸纤维素滤膜和滤纸装入过滤装置，连接蠕动泵，过滤污水。待过滤结束后，在生物安全柜中拆开过滤装置，取出硝酸纤维素滤膜放到无菌平皿中，用无菌剪刀和无菌镊子将滤膜剪碎，放入50 mL离心管中。向离心管中加入10 mL的3%牛肉浸出液，加入玻璃珠，在振荡器上480 rpm振荡30 min，然后于3 000 rpm下4℃离心30 min，收集上清液用针式滤器（0.22 μm）过滤至15 mL的离心管中，进行后续实验。
1.2.2　肠道病毒分离和鉴定
肠道病毒分离时所用的转入脊髓灰质炎受体的小鼠肺细胞系（Mouse Cell Line Expressing the Gene for the Human Cellular Receptor for Poliovirus, L20B）、人横纹肌肉瘤细胞（Human Rhabdomyosarcoma, RD）由中国疾病预防控制中心-病毒病预防控制所-国家脊髓灰质炎实验室提供，病毒的分离鉴定操作程序根据《脊灰实验室操作手册》（世界卫生组织WHO第4版）的标准操作规程进行[13]。每份污水样本在每种细胞的24孔板上接种8孔。观察CPE，收获阳性分离物并进行型内鉴定。型内鉴定使用世界卫生组织下发的Poliovirus rRT-PCR ITD 5.1 kit试剂盒、Poliovirus rRT-PCR VDPV 5.0 kit试剂盒和美国Promega的GoTaq© Probe 1-Step RT-qPCR System试剂盒。型内鉴定使用美国ABI公司的7500 Real Time PCR System，严格遵照相关操作说明进行实验。
1.2.3　核酸提取
病毒的总核酸提取采用西安天隆科技有限公司的全自动核酸提取仪（GeneRotex_96），试剂采用该公司的病毒DNA/RNA提取试剂盒（PLUS），按操作说明进行。
1.2.4　腹泻病毒核酸检测
腹泻病毒的核酸检测使用硕世生物的诺如病毒GⅠ/GⅡ型和轮状病毒A组核酸检测试剂盒（荧光PCR法），札如病毒、星状病毒和肠道腺病毒核酸检测试剂盒（荧光PCR法）。荧光PCR反应使用宏石公司的SLAN-96S Real-Time PCR System，严格遵照相关操作说明进行实验。
2　结果
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2.1　肠道病毒分离情况
2022年1—12月共采集12份环境污水，共分离出脊髓灰质炎病毒3株和非脊灰肠道病毒9株。在4月和5月分离到脊髓灰质炎病毒，而非脊灰肠道病毒在2-6月均有分离（表1）。
2.2　脊灰病毒鉴定结果
对分离到的3株脊髓灰质炎病毒进行型内鉴定，结果均为Ⅲ型脊灰疫苗相似株，未发现脊灰疫苗衍生病毒（Vaccine-Derived Poliovirus, VDPV）或脊灰野毒株（Wild Poliovirus, WPV）。
2.3　腹泻病毒核酸检测情况
2022年1—12月的环境污水中，每月都有A组轮状病毒和诺如病毒GⅡ型的检出；诺如病毒GⅠ型和肠道腺病毒除11月外其他月份均有检出；人星状病毒在7月、8月、11月、12月未能检出；札如病毒全年未检出（表1）。
3　讨论
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肠道病毒感染以隐性感染为主，只有少数感染者（约1%左右）会出现相关临床症状，隐性感染者难以发现[13]。荷兰在1992年应对一次Ⅲ型脊灰病例的暴发疫情中观察到：生活污水中发现脊灰病毒的时间早于同源脊灰病毒导致的首例指示病例[14]，说明在首例指示病例出现症状前，脊灰病毒已在生活污水和隐性感染人群中出现，提示对环境脊灰病毒的长期检测可助力脊灰病毒暴发预警[15]。阿坝州曾于2012年发生过Ⅱ型脊灰疫苗衍生病毒循环事件（cVDPVs），作为重点关注地区，采用环境监测能够更加真实地反应环境中肠道病毒尤其是脊灰病毒的存在和循环情况。本研究共分离到Ⅲ型脊灰疫苗相似株3株，未发现脊灰野毒株或脊灰疫苗衍生病毒，说明脊灰病毒在该地区未出现高变异病毒也未在人群中发生循环。本次试验仅对肠道病毒中的脊灰病毒进行了深入研究，对非脊灰肠道病毒还未进一步分析，今后有待于开展相关的研究。
近年来，关于A组轮状病毒、诺如病毒等腹泻病毒导致的病毒性胃肠炎疾病的报告越来越多，已成为危害严重的公共卫生问题之一[16-17]。并且各病毒型别众多、进化活跃，在不同时期、不同地区和不同人群中的流行情况存在较大差异[10, 18]。目前我国腹泻病毒的监测多集中在住院病例和暴发病例监测，存在一定选择偏倚，无法全面反映研究地区腹泻病毒的动态变迁和局部扩散情况。环境污水监测不仅能全面反映研究地区腹泻病毒的分子特征，并且可以分析和预测这些病毒的时间分布和变异趋势。本研究发现，在阿坝州环境样本中A组轮状病毒、诺如病毒GⅡ型、诺如病毒GⅠ型、肠道腺病毒和人星状病毒检出率均很高，这与山东、成都、广州地区的结果是一致的[5，19-21]。关于札如病毒的环境污水方面的研究比较少，但本次研究中札如病毒的检出情况与2011年-2020年成都市腹泻儿童病例的研究结果是相似的[10]，人群中札如病毒检出率也较低。本次试验说明在阿坝州流行的腹泻病毒主要为A组轮状病毒和诺如病毒，肠道腺病毒和人星状病毒存在散发，札如病毒未在阿坝州流行。
时间分布方面，我国病毒感染主要在冬春季节，从头年12月起到次年5月结束；细菌性感染主要在夏秋季节，从5月起到10月结束[22-23]。本研究发现，阿坝地区污水中肠道病毒的非脊灰肠道病毒分离检出时间为2—6月，脊灰病毒分离株检出时间为4—5月。肠道病毒检出时间段与上述研究的病毒感染时间大致吻合；但腹泻病毒全年均有检出，与上述研究的病毒感染时间并不一致，推测可能与当地低纬度高海拔高山峡谷立体气候及藏羌少数民族聚居特性有关，有待进一步研究。
开展环境污水的肠道病毒和腹泻病毒监测，可作为病例监测的重要补充，为在人群中的肠道病毒和腹泻病毒相关疾病的流行提供了预测预警的关键作用，将传染病应对关口前移，体现了预防为主的卫生方针政策。
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2024年第51卷第20期
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doi: 10.20043/j.cnki.MPM.202312247
  • 接收时间：2023-12-15
  • 首发时间：2026-03-20
  • 出版时间：2024-10-25
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  • 收稿日期：2023-12-15
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    1.阿坝州疾病预防控制中心，四川 马尔康 624050
    2.四川省疾病预防控制中心

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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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