胡慧-新發豬病毒性腹瀉的流行特點及防控措施.pdf

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1、新發豬病毒性腹瀉的流行特點及防控措施Epidemiological Characteristics and Prevention and Control Measures of Newly Emerging Porcine Viral Diarrhea胡 慧Hu Hui河南農業大學/動物醫學院Henan Agricultural University/School of Veterinary Medicine2024.4.26 河南鄭州April 26,2024 Zhengzhou,Henan一一帶帶一一路路國國際際生生豬豬產產業業大大會會Belt and Road International

2、 Swine Industry Confe一帶一路國際生豬產業大會二新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology of Newly Emerging Porcine Deltacoronavirus(PDCoV)And Infection Characteristics目目 錄錄CONTENTS四新發豬病毒性腹瀉防控技術研究Research on Prevention and Control Techniques of Newly Emerging Porcine Viral Diarrhea三其它豬病毒性腹瀉病原的發現Discovery of Other Patho

3、gens of Porcine Viral Diarrhea一豬病毒性腹瀉概述Overview of Porcine Viral Diarrhea一帶一路國際生豬產業大會一、豬病毒性腹瀉概述I.Overview of Porcine Viral Diarrhea仔豬腹瀉Piglet diarrhea病毒性腹瀉Viral diarrhea豬豬流流行行性性腹腹瀉瀉Porcine epidemic diarrhea輪輪狀狀病病毒毒感感染染Rotavirus infection豬豬冠冠狀狀病病毒毒感感染染Porcine deltacoronavirus infection細菌性腹瀉Bacterial

4、diarrhea仔仔豬豬紅紅、白白痢痢Piglet red and white dysentery副副傷傷寒寒Paratyphoid fever回回腸腸炎炎Ileitis慢慢性性腸腸炎炎Chronic enteritis生理性腹瀉Physiological diarrhea斷斷奶奶應應激激反反應應引引起起的的腸腸炎炎、腹腹瀉瀉、水水樣樣拉拉稀稀等等Enteritis,diarrhea,watery stools caused by weaning stress response etc.母源性腹瀉Maternal Diarrhea前前期期黃黃白白色色糊糊狀狀糞糞便便;Early-stage y

5、ellowish-white pasty feces中中期期水水瀉瀉;Mid-stage watery diarrhea后后期期脂脂肪肪性性腹腹瀉瀉Late-stage fatty diarrhea霉菌性腹瀉Mycotoxins diarrhea各各種種霉霉菌菌毒毒素素引引起起的的下下痢痢、嘔嘔吐吐、脫脫肛肛及及慢慢性性胃胃腸腸炎炎Dysentery,vomiting,prolapse,and chronic gastroenteritis caused by various mycotoxins仔豬腹瀉的病原種類繁多、原因復雜Piglet diarrhea is caused by a va

6、riety of pathogens and complex reasons仔豬生長受阻、高死亡率Piglet growth retardation,high mortality rate一帶一路國際生豬產業大會一、豬病毒性腹瀉概述I.Overview of Porcine Viral Diarrhea 豬傳染性胃腸炎病毒 Transmissible Gastroenteritis Virus of Swine（TGEV）豬冠狀病毒Porcine Deltacoronavirus（PDCoV）2010年年1946年年2014年年2017年年豬流行性腹瀉病毒Porcine Epidemic Di

7、arrhea Virus（PEDV）豬急性腹瀉綜合征冠狀病毒Swine Acute Diarrhea Syndrome Coronavirus（SADS-CoV）1982年年豬輪狀病毒Porcine Rotavirus（PoRV）豬薩佩羅病毒(PSV)Porcine Sapelovirus(PSV)豬星狀病毒(PAstV)Porcine Astrovirus(PAstV)豬呼腸孤病毒(PRV)Porcine Reovirus(PRV)新發與再現的豬病毒性腹瀉頻繁發生The newly emerging and recurring cases of porcine viral diarrhea

8、occur frequently.豬病毒性腹瀉是困擾我國乃至全球養豬業的突出問題Porcine viral diarrhea is a prominent issue affecting the swine industry in China and globally.一帶一路國際生豬產業大會一、豬病毒性腹瀉概述I.Overview of Porcine Viral Diarrhea豬病毒性腹瀉的危害The hazards of porcine viral diarrhea危危害害Hazards降低Decrease升高Increase仔豬死亡率升高，Piglet mortality rates

9、 increased,養豬成本升高，And pig farming costs elevated,出現“沒豬養”的困境。Leading to a dilemma of insufficient pig farming.養豬效益降低，威脅養豬業的健康發展、食品安全、公共衛生安全。The declining profitability of pig farming threatens the healthy development of the industry,food safety,and public health.一帶一路國際生豬產業大會1.豬冠狀病毒（PDCoV）簡介 新發冠狀病毒1.P

10、orcine Deltacoronavirus(PDCoV)Introduction Newly Emerging Coronavirus二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)1冠狀病毒（SARS-CoV、MERS-CoV、SARS-CoV-2)Coronaviruses(SARS-CoV,MERS-CoV,SARS-CoV-2)2冠狀病毒（禽類CoV如IBV、TCoV)Coronaviruses(A

11、vian CoVs such as IBV,TCoV)3冠狀病毒（Bird CoV、PDCoV)Coronaviruses(Bird CoV,PDCoV)4冠狀病毒（人冠狀病毒、PEDV、TGEV、SeACoV等）Coronaviruses(Human coronaviruses,PEDV,TGEV,SeACoV,etc.)Coronavirus一帶一路國際生豬產業大會PDCoV是2014年新發現的仔豬腹瀉病PDCoV is a newly discovered cause of piglet diarrhea in 20142022年6月中國農業農村部將豬丁型冠狀病毒感染列為三類動物疫病In

12、 June 2022,the Chinese Ministry of Agriculture and Rural Affairs classified porcine deltacoronavirus infection as a Category III animal disease確確切切時時間間？Exact timeline?起起源源？Origin?宿宿主主？Host?進進化化？Evolution?2.PDCoV的發現 Discovery of PDCoV 二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristic

13、s of Newly Emerging Porcine Deltacoronavirus(PDCoV)一帶一路國際生豬產業大會 Kim et al.,Pathogens,2023美美國國PDCoV流流行行情情況況The situation of PDCoV in the United StatesKikuti et al.,Viruses,2024 二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)目前呈全球流行

14、It is currently prevalent worldwide2014年美國豬場首次暴發PDCoV，持續存在，有上升趨勢The first outbreak of PDCoV in US pig farms occurred in 2014 and has since persisted,showing an upward trend東南亞流行毒株致病性較強Strains circulating in Southeast Asia exhibit higher pathogenicity3.PDCoV的流行情況全球流行情況3.Epidemiology of PDCoV Global S

15、ituation一帶一路國際生豬產業大會Li et al.,Bioscience Reports,2019;Zhang et al.,Preventive Veterinary Medicine,20192015年開始豬場陸續檢出PDCoVSince 2015,PDCoV has been gradually detected in pig farms worldwide.核酸陽性率在20-30%The nucleic acid positive rate ranges from 20%to 30%抗體陽性率在60%左右With an antibody positive rate of aro

16、und 60%混合感染普遍（PEDV/PoRV/PSV等）Co-infections are common(e.g.,PEDV/PoRV/PSV)一年四季均可發病，冬季節多發The disease can occur throughout the year,with a higher incidence during winter3.PDCoV的流行情況中國流行情況3.Epidemiology of PDCoV Prevalence situation in ChinaPDCoV 流行呈逐年上升趨勢！The prevalence of PDCoV is increasing year by y

17、ear in China!二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)一帶一路國際生豬產業大會PDCoV可以感染各年齡段的豬只；主要引起15日齡以內仔豬發生嚴重腹瀉、新生仔豬死亡；母豬可感染，多表現一過性腹瀉癥狀。PDCoV can infect pigs of all age groups;it mainly causes severe diarrhea in piglets under 15 days

18、old,leading to neonatal piglet deaths.Sows can also be infected,often exhibiting transient diarrhea symptoms.PDCoV感感染染引引起起全全身身炎炎癥癥和和多多組組織織病病理理損損傷傷，PDCoV infection causes systemic inflammation and multi-organ pathological damage,破破壞壞仔仔豬豬腸腸道道結結構構，仔仔豬豬小小腸腸絨絨毛毛嚴嚴重重受受損損。Disrupting piglet intestinal struc

19、ture and severely damaging intestinal villi.Jejunum空空腸腸Ileum回回腸腸4.PDCoV對豬的致病性可感染不同年齡豬只4.The Pathogenicity of PDCoV to pigs its ability to infect of pigs of all ages二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)一帶一路國際生豬產業大會PDCoV自然

20、感染仔豬PDCoV naturally infected pigletsPDCoV人工感染仔豬PDCoV artificial Infected PigletsPDCoV自然感染母豬PDCoV naturally infected sows二、新發豬冠狀病毒的流行情況及感染特性研究II.The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)4.PDCoV對豬的致病性多組織嗜性特點4.The Pathogenicity of PDCoV in Pigs-Cha

21、racteristics of Multi-Tissue Tropism可在感染豬只的腸道、腎、肺、肝、脾等組織器官中檢出病毒；The virus can be detected in the intestines,kidneys,lungs,liver,spleen and other tissues and organs of infected pigs;仔豬腸系膜淋巴結、腹股溝淋巴結檢出病毒，具有多組織嗜性。The virus is detected in the mesenteric lymph nodes and inguinal lymph nodes of piglets,sho

22、wing its characteristics of multi-tissue tropism.一帶一路國際生豬產業大會臨床上PEDV和PDCoV共感染普遍In clinical practice,co-infection of PEDV and PDCoV is common.PEDV疫苗株CV777促進PDCoV感染CV777,the vaccine strain of PEDV,promotes the infection of PDCoV二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of New

23、ly Emerging Porcine Deltacoronavirus(PDCoV)4.PDCoV對豬的致病性協同致病特性4.The Pathogenicity of PDCoV in pigs-Characteristics of synergistic pathogenicityZhang et al.,Transbound Emerg Dis,2019Guo et al.,Emerg Microbes Infect,2024一帶一路國際生豬產業大會對PDCoV S基因遺傳進化分析發現PDCoV與亞洲豹貓的-CoV親緣關系最近；The genetic evolutionary analy

24、sis of the PDCoVS gene reveals that PDCoV is closest to the-CoV of the Asian leopard cat;對PDCoV全基因序列分析表明PDCoV與麻雀來源的-CoV親緣關系最近。The analysis of the whole gene sequence of PDCoV shows that PDCoV has the closest relationship with-CoV from sparrow.二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Chara

25、cteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)5.PDCoV具有跨種感染特性5.PDCoV has the characteristic of cross-species infection.提示其跨種傳播潛力！It suggests the potential of cross-species transmission!一帶一路國際生豬產業大會PDCoV感染SPF雞后有輕微腹瀉和腸道損傷情況，表現為多組織嗜性。SPF chickens infected with PDCoV have mild diarrhea and

26、 intestinal damage,which shows its characteristics of multiple tissue tropism.PDCoV可以人工感染不同日齡的雞！Chicken of different ages can be artificially infected with PDCoV!Liang et al.,2019,Viruses二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PD

27、CoV)5.PDCoV具有跨種感染特性感染家禽5.PDCoV has the characteristic of cross-species infection-infecting poultry.一帶一路國際生豬產業大會PDCoV可以人工感染新生無菌犢牛！Neonatal aseptic calves can be artificially infected with PDCoV!二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavi

28、rus(PDCoV)5.PDCoV具有跨種感染特性感染犢牛5.PDCoV has the characteristic of cross-species infection-infecting calvesPDCoV感染犢牛后糞便中排毒時間和人工感染仔豬的一致；血清中檢測到特異性IgG抗體；牛無臨床癥狀，剖檢無變化。The shedding time of PDCoV in the feces of infected calves is the same as that of artificially infected piglets;specific IgG antibody is dete

29、cted in the serum;no clinical symptoms are found in calves,and there is no change in necropsy.Jung et al.,Arch Virol,2017一帶一路國際生豬產業大會*Zhang et al.,Journal of Medical Virology,2022二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)5.PD

30、CoV具有跨種感染特性感染小鼠5.PDCoV has the characteristic of cross-species infection-infecting micePDCoV感染小鼠后有明顯的腸道損傷，表現為多組織嗜性，產生特異性中和抗體。Mice infected with PDCoV has obvious intestinal damage,showing the characteristic of multi-tissue tropism,and mice produce specific neutralizing antibodies.一帶一路國際生豬產業大會二、新發豬冠狀

31、病毒的流行情況及感染特性研究II.The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)5.PDCoV具有跨種感染特性具有感染人的能力5.PDCoV has the characteristic of cross-species infection-infecting humans具備感染人的潛能：The potential to infect humans:可感染7個種屬的20余種細胞系（包括人源細胞系）；It can infect more than 2

32、0 cell lines of 7 species(including human cell lines);PDCoV的RBD與人和豬的APN結合區域結構高度相似，可能是其感染豬和人的結構基礎。The RBD of PDCoV has a highly similar structure to the binding region of APN in humans and pigs,which may serve as the structural basis for its infection of both pigs and humans.跨種感染人：2021年變異的PDCoV感染海地的3

33、名兒童。Cross-species infection in humans:Three children in Haiti infected with a mutant PDCoV in 2021.一帶一路國際生豬產業大會 Where did PDCoV come from?（PDCoV從從哪哪里里來來？）Where is PDCoV going?（PDCoV會會到到哪哪里里去去？）Viral fitness and pathogenicity of PDCoV in different host （PDCoV的的傳傳播播適適應應性性和和致致病病性性變變化化趨趨勢勢）持續監測 ongoing

34、monitoring二、新發豬冠狀病毒的流行情況及感染特性研究The Epidemiology and Infection Characteristics of Newly Emerging Porcine Deltacoronavirus(PDCoV)PDCoV我們更關注的問題：What we focus on:一帶一路國際生豬產業大會Son,et al.,Front Microbiol,2019三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogens1.豬薩佩羅病毒（PSV）1.Porcine Sapelo

35、virus(PSV)PSV是無囊膜的單股正鏈RNA病毒，屬于小RNA病毒科薩佩羅病毒屬的成員，可感染野豬和不同年齡段的家豬，主要引起急性腹瀉、肺炎、腦脊髓炎和生殖障礙等癥狀；PSV is a single-stranded positive-stranded RNA virus without envelope,which belongs to the PSV genus,picornaviridae family.PSV can infect wild boars and domestic pigs of different ages,mainly causing these symptom

36、s such as acute diarrhea,pneumonia,encephalomyelitis and reproductive disorders;本團隊從腹瀉豬病料中分離到2株PSV，分別為HNHB株和HNXY株。Two PSV strains,HNHB and HNXY,were isolated from diarrheal swine samples by our team.Zhang.,et al.,Transbound Emerg Dis,2022Unpublished data未未公公布布數數據據一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Disco

37、very of other porcine viral diarrhea pathogens1.豬薩佩羅病毒（PSV）1.Porcine Sapelovirus(PSV)Zhang.,et al.,Transbound Emerg Dis,2022Unpublished data未未公公布布數數據據PSV可感染不同日齡豬，引起5日齡仔豬發生腹瀉、抽搐等癥狀，引起腸道組織損傷；PSV can infect pigs at different ages,and lead to diarrhea,convulsions and other symptoms in 5-day-old piglets,

38、thus causing intestinal tissue damage;PSV具有協同致病作用。通過建立PSV和PDCoV共感染仔豬模型，發現混合感染可加重腸道絨毛損傷，PDCoV促進了PSV在體內的增殖，而PSV抑制PDCoV的增殖。PSV has the effect of synergistic pathogenesis.By establishing a piglet model co-infected with PSV and PDCoV,it is found that mixed infection could aggravate the damage of intestin

39、al villi.PDCoV promotes the proliferation of PSV in vivo,while PSV inhibits the proliferation of PDCoV.一帶一路國際生豬產業大會地地區區Areas陽陽性性率率Positive rate國外Overseas西班牙Spain9%捷克Czech32.3%韓國South Korea34%印度India5.33%國內Domestic華東East China17.2%華南South China18.2%四川Sichuan26.6%三、其它豬病毒性腹瀉病原的發現III.Discovery of other

40、porcine viral diarrhea pathogens1.豬薩佩羅病毒（PSV）1.Porcine Sapelovirus(PSV)PSV在多個國家豬群中均檢測到，陽性率高達34%；PSV has been detected in pig herds in many countries,with a positivity rate as high as 34%；PSV在我國多個省份流行，多與其它病毒共感染，陽性率達30%。PSV is prevalent in many provinces in China,mostly co-infected with other viruses,

41、with a positive rate of 30%.一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogens2.哺乳動物呼腸孤病毒（MRV）2.Mammalian Reovirus(MRV)屬于呼腸孤病毒科正呼腸孤病毒屬，是一種雙鏈RNA病毒，宿主譜非常廣泛，主要引起呼吸道和腸道感染；It belongs to the Orthoreovirus genus of the Reoviridae family.It is a double-stranded RNA virus with

42、 a wide range of hosts,mainly causing respiratory and intestinal infections;本團隊從腹瀉豬病料中分離到一株MRV，命名為HN2019-01；We isolated a strain of MRV from diarrheal swine samples,named HN2019-01;MRV可以在多種細胞上復制，且外源蛋白酶的添加有助于病毒生長。MRV can replicate on a variety of cells,and the addition of exogenous proteases contribu

43、tes to viral growth.Unpublished data未未公公布布數數據據一帶一路國際生豬產業大會MRV感染5日齡仔豬后引起腹瀉癥狀不明顯，腸道組織有損傷；Infection with MRV in 5-day-old piglets does not cause obvious diarrhea symptoms,but damages the intestinal tissues.MRV破壞仔豬的腸道菌群的豐度和結構，有害菌顯著上調；MRV destroys the abundance and structure of intestinal microflora in p

44、iglets,with harmful bacteria significantly up-regulated;MRV的協同致病情況尚不確定。The synergistic pathogenesis of MRV is uncertain.三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogens2.哺乳動物呼腸孤病毒（MRV）2.Mammalian Reovirus(MRV)Unpublished data未未公公布布數數據據一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Discovery of

45、 other porcine viral diarrhea pathogens2.哺乳動物呼腸孤病毒（MRV）2.Mammalian Reovirus(MRV)在2015-2016年從山東、河南、江西、黑龍江和浙江等省共25個豬場收集到的262份腹瀉豬血清樣品中，抗體陽性率達84.4%。Among the 262 serum samples of diarrhea pigs collected from 25 pig farms in Shandong,Henan,Jiangxi,Heilongjiang and Zhejiang provinces from 2015 to 2016,the

46、 positive rate of antibody reached 84.4%.覃盼等，浙江大學學報，2017Qin Pan et al,Journal of Zhejiang University,2017一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogensLi.,et al.TransBoundary Eme Dis.20243.豬星狀病毒（PAstV)3.Porcine Astrovirus(PAstV)PAstV屬于星狀病毒科哺乳動物星狀病毒屬，是一種無囊膜的單股正鏈RNA

47、病毒，存在5種基因型，可感染不同年齡段的家豬，主要引起仔豬腹瀉等癥狀；PAstV belongs to the Mammalian Astrovirus genus of the Astroviridae family.It is a non-enveloped single-stranded positive-strand RNA virus with five genotypes.It can infect domestic pigs of different ages and mainly cause diarrhea and other symptoms in piglets;本團隊從

48、腹瀉豬病料中分離到一株PAstV5型流行毒株（HNPDS株）。An AstV5 epidemic strain(named HNPDS strain)is isolated from diarrheic pig samples by our team.一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogensLi.,et al.TransBoundary Eme Dis.20243.豬星狀病毒（PAstV)3.Porcine Astrovirus(PAstV)PAstV 5感染引起仔豬輕微

49、腹瀉，生長速度減慢，腸道絨毛萎縮及脫落。PAstV 5 infection causes mild diarrhea,slowed growth rate,and atrophy and shedding of intestinal villi in piglets.PAstV 5引起回腸中的淋巴濾泡增大及輪廓模糊。PAstV 5 causes enlarged lymphoid follicles and blurred contours in the ileum.PAstV 5感染會破壞仔豬的腸道菌群的豐度及微生物群落組成。PAstV 5 infection disrupts the ab

50、undance of gut microbiota and microbial community composition in piglets.一帶一路國際生豬產業大會不不同同基基因因型型PAstV在在中中國國的的流流行行情情況況The prevalence of different genotypes of PAstV in China三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogens中中國國PAstV在在不不同同年年份份的的陽陽性性率率The positive rates of PAstV in d

51、ifferent years in China中中國國PAstV在在不不同同地地區區的的陽陽性性率率The positive rates of PAstV in different areas in China周以鐵,等,中國預防獸醫學報Zhou Yitie et al.,Chinese Journal of Preventive Vet3.豬星狀病毒（PAstV)3.Porcine Astrovirus(PAstV)2006 年2021 年間，PAstV 總陽性率為 27.45%（1467/5344）。Between 2006 and 2021,the total positive rate

52、 of PAstV was 27.45%(1467/5344).2019-2021年PAstV檢出率為34.5%，主要為PAstV 2及PAstV 5兩種基因型。Between 2019 and 2021,the detection rate of PAstV was 34.5%,mainly consisting of two genotypes:PAstV 2 and PAstV 5.在腹瀉及健康樣本中均有檢出，腹瀉樣本中陽性率為28.8%，健康樣本中陽性率為17.1%。The detection samples comprise both diarrhea and healthy sam

53、ples,with a positive rate of 28.8%in diarrhea samples and 17.1%in healthy samples.一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogens4.變異的豬傳染性胃腸炎病毒（vTGEV）4.Variant Transmissible Gastroenteritis Virus(vTGEV)免疫壓力和病毒自身變異導致臨床上變異的TGEV毒株不斷出現；Immune pressure and virus mutatio

54、n lead to the continuous emergence of vTGEV strains in clinical practice;TGEV的基因突變多集中在S和ORF3a基因，多為基因片段的缺失。The genetic mutations of TGEV are mostly concentrated in the S and ORF3a genes,mostly due to the deletion of gene fragments.2018-2021年年 中中國國中中南南部部，Li et al.,2022From 2018 to 2021 Central and Sou

55、thern China,Li et al.,2022一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogens4.變異的豬傳染性胃腸炎病毒（vTGEV）4.Variant Transmissible Gastroenteritis Virus(vTGEV)本實驗室于2012年檢測并分離到TGEV變異毒株HN-2012，缺失ORF3a基因；In 2012,our laboratory detected and isolated the TGEV variant strain HN-2012,

56、which lacked the ORF3a gene;缺失位點和2015年報道的TGEV毒株位置有差異，但仍然屬于Miller譜系。The missing site differs from the TGEV strain reported in 2015,but still belongs to the Miller lineage.一帶一路國際生豬產業大會三、其它豬病毒性腹瀉病原的發現III.Discovery of other porcine viral diarrhea pathogens4.變異的豬傳染性胃腸炎病毒（vTGEV）4.Variant Transmissible Gas

57、troenteritis Virus(vTGEV)TGEV變異毒株HN-2012（缺失ORF3基因）組織嗜性發生變化，感染仔豬后腹瀉不明顯，腸道、肺部、腦部病變明顯；The tissue tropism of the TGEV variant strain HN-2012(ORF3 gene deleted)changes,and the diarrhea of piglets after infection is not obvious,but there are obvious lesions in the intestines,lungs,and brain;利用反向遺傳技術拯救TGEV

58、-ORF3毒株，相較于TGEV HN-2012組，rTGEV-ORF3對仔豬致病力降低，在仔豬體內的復制能力降低。We can use reverse genetic technology to save TGEV-ORF3 strain.Compared to the TGEV HN-2012 group,rTGEV-ORF3 has a reduced pathogenicity to piglets and a reduced replication ability in piglets.一帶一路國際生豬產業大會四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevent

59、ion and Control Techniques for Emerging Porcine Viral Diarrhea豬病毒性腹瀉病原復雜，疫病防控形勢嚴峻The pathogen of porcine viral diarrhea is complex,and the situation of disease prevention and control is severe面臨的“卡脖子”問題The bottleneck problems faced一帶一路國際生豬產業大會p針對豬病毒性腹瀉新病頻發、臨床癥狀相似等問題，建立了實驗室精準診斷、鑒別診斷和臨床快速診斷技術，創建了豬腹瀉病原

60、快速檢測技術平臺，開發快速檢測技術共20項。To address the frequent occurrence of new diseases of porcine viral diarrhea and similar clinical symptoms,we have established laboratory precision diagnosis,differential diagnosis,clinical rapid diagnosis technologies,created a rapid detection technology platform for porcine d

61、iarrhea pathogens and developed a total of 20 rapid detection technologies.快速檢測技術的建立The establishment of rapid detection technologyDNA四面體檢測DNA tetrahedral detectiondd-PCR檢測dd-PCR detection免疫磁珠法IMB膠體金檢測Colloidal gold detection競爭ELISACompetitive ELISA間接ELISAIndirect ELISA抗體檢測Antibody testing抗原檢測Antige

62、n testing核酸檢測Nucleic acid testing特異性診斷標識的挖掘和制備Mining and preparation of specific diagnostic markers蛋白診斷標準品的制備Preparation of protein diagnostic standards核酸診斷標準品的制備Preparation of nuclear acid diagnostic standards挖掘診斷標識Mining diagnostic markers1.新發豬病毒性腹瀉診斷技術研究1.Research on Diagnostic Techniques for Eme

63、rging Porcine Viral Diarrhea診斷產品制備Preparation of diagnostic products四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea一帶一路國際生豬產業大會正正向向引引物物設設計計區區域域Forward primer design region反反向向引引物物設設計計區區域域Reverse primer design region能 夠 檢 測 已 知 所 有 的 冠 狀 病 毒，Capabl

64、e of detecting all known coronaviruses,和 未 來 可 能 新 出 現 的 冠 狀 病 毒。and potentially new coronaviruses that may emerge in the future.1.新發豬病毒性腹瀉診斷技術研究1.Research on Diagnostic Techniques for Emerging Porcine Viral Diarrheap針對新的冠狀病毒頻發現象，首創冠狀病毒通用qRT-PCR檢測技術，可用于冠狀病毒的溯源、宿主譜的挖掘和對未來暴發的冠狀病毒進行預警。In response to th

65、e frequent occurrence of new coronaviruses,we have pioneered the universal qRT PCR detection technology for coronaviruses,which can be used for tracing the origin of coronaviruses,mining host range,and providing early warning for future outbreaks of coronaviruses.四、新發豬病毒性腹瀉防控技術研究IV.Research on Preve

66、ntion and Control Techniques for Emerging Porcine Viral Diarrhea一帶一路國際生豬產業大會u鑒別診斷：同時檢測多種病原，確診時間降至2h；Differential diagnosis:simultaneous detection of multiple pathogens,with diagnosis time reduced to 2 hours;u早期診斷：膠體金試紙條最短5min確診，數字PCR技術將檢測精度提高100倍；Early diagnosis:The colloidal gold test strip takes a

67、 minimum of 5 minutes for diagnosis,and digital PCR technology improves detection accuracy by 100 times;u通用檢測：可檢測已知所有的冠狀病毒，用于冠狀病毒溯源和新發冠狀病毒的預警、監測。Universal detection:detection of all known coronaviruses,used for coronavirus tracing and early warning and monitoring of emerging coronaviruses.類別Classifi

68、cation技術名稱Technical Name應用Application時間Time精度Precision專利Patent豬德爾塔冠狀病毒和豬流行性腹瀉病毒多重RT-PCR檢測引物及檢測方法（ZL201511016093.8）Multiple RT-PCR detection primers and detection methods for Porcine Delta Coronavirus and Porcine Epidemic Diarrhea Virus(ZL201511016093.8)鑒別診斷Differential diagnosis2h高（102copy/L）High(10

69、2copy/L)豬德爾塔冠狀病毒、豬流行性腹瀉病毒、豬傳染性胃腸炎病毒多重RT-PCR檢測引物（ZL201511021732.X）Multiple RT-PCR detection primers for Porcine Delta Coronavirus,Porcine Epidemic Diarrhea Virus,and Porcine Transmissible Gastroenteritis Virus(ZL201511021732.X)鑒別診斷Differential diagnosis2h高（102copy/L）High(102copy/L)豬德爾塔冠狀病毒和豬傳染性胃腸炎病毒多

70、重RT-PCR檢測引物及檢測方法（ZL201511025198.X）Multiple RT-PCR detection primers and detection methods for Porcine Delta Coronavirus and Porcine Transmissible Gastroenteritis Virus(ZL201511025198.X)鑒別診斷Differential diagnosis2h高（102copy/L）High(102copy/L)一種檢測豬德爾塔冠狀病毒和豬流行性腹瀉病毒的微滴式雙重數字PCR試劑盒（ZL202211561712.1）A Micro

71、droplet Dual Digital PCR Kit for Detecting Porcine Delta Coronavirus and Porcine Epidemic Diarrhea Virus(ZL202211561712.1)鑒別診斷Differential diagnosis2h極高（1copy/L）Extremely high(1copy/L)一種用于檢測豬薩佩羅病毒的芯片式數字 PCR 試劑盒（ZL 2023 1 0174664.9）A chip based digital PCR kit for detecting porcine Sapelovirus(ZL 202

72、3 1 0174664.9)早期診斷Early diagnosis2h極高（1copy/L）Extremely high(1copy/L)標準Standard豬流行性腹瀉病毒RT-PCR檢測方法（GB/T34757-2017）RT-PCR detection method for porcine epidemic diarrhea virus(GB/T34757-2017)早期診斷Early diagnosis2h高（102copy/L）High(102copy/L)豬傳染性胃腸炎病毒RT-nPCR檢測方法RT-nPCR detection method for porcine transmi

73、ssible gastroenteritis virus（NY/T 2841-2015）早期診斷Early diagnosis2h高（102copy/L）High(102copy/L)豬丁型冠狀病毒病診斷技術（NYB-23099）Diagnostic techniques for porcine D-type coronavirus diagnostic technology(NYB-23099)早期診斷Early diagnosis2h高（102copy/L）High(102copy/L)Hu H et al.,J Virol Methods.2018；丁慶文等，中國預防獸醫學報，2021；

74、Ding qingwen et al.,Chinese Journal of Preventive Veterinary Medicine,2021;閆瑞杰等，西北農林科技大學學報，2019；Yan Ruijie et al.,Journal of Northwest A&F University,2019;張利衛等，中國獸醫學報，2018；Zhang Liwei et al.,Chinese Journal of Veterinary Medicine,2018;韋學雷等，中國獸醫學報，2018；Wei Xuelei et al.,Chinese Journal of Veterinary

75、Medicine,2018;劉玲玲等，農業生物技術學報，2016 Liu Lingling et al.,Journal of Agricultural Biotechnology,2016.1.新發豬病毒性腹瀉診斷技術研究1.Research on Diagnostic Techniques for Emerging Porcine Viral Diarrhea四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea一帶一路國際生豬產業大會2.新發豬

76、病毒性腹瀉疫苗研究滅活疫苗2.Research on Vaccines for Newly Emerged Porcine Viral Diarrhea-Inactivated VaccinesZheng et al.,Vaccine,2022p針對一些新發疫病無疫苗的問題，開展了PDCoV、PSV滅活疫苗的研制，攻克該類病毒無疫苗可用的產業難題，實現疫病的精準、高效防控。To address the issue of the lack of vaccines for some newly emerged diseases,research has been conducted on the

77、development of inactivated vaccines for PDCoV and PSV to overcome the industrial challenge of the unavailability of vaccines for these viruses,enabling precise and efficient prevention and control of the diseases.pPDCoV滅活疫苗已經進入臨床批件申報，PSV滅活疫苗已經完成實驗室研究。The inactivated vaccine for PDCoV has entered the

78、 clinical application approval process,while the inactivated vaccine for PSV has completed laboratory research.四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea一帶一路國際生豬產業大會2.新發豬病毒性腹瀉疫苗研究弱毒疫苗2.Research on Vaccines for Newly Emerged Porcine Viral Diar

79、rhea-MLVUnpublished data未未公公布布數數據據Zhang et al.,msystem,2024p對PDCoV、TGEV和PSV進行連續傳代，篩選出復制能力強、遺傳特性穩定、致病力降低的毒株，為弱毒疫苗研發提供合適的候選毒株。Through continuous passages of PDCoV,TGEV,and PSV,strains with strong replication ability,stable genetic characteristics,and reduced pathogenicity have been screened out.These

80、strains provide suitable candidates for the development of MLV.四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea一帶一路國際生豬產業大會2.新發豬病毒性腹瀉疫苗研究反向遺傳學疫苗2.Research on Vaccines for Newly Emerged Porcine Viral Diarrhea-Reverse Genetics VaccinesZhang et al.,J.

81、Agric.Food.Chem,2024p針對傳統疫苗研制過程中，候選毒株篩選難等問題，利用酵母轉化偶聯重組技術建立豬腸道冠狀病毒反向遺傳操作技術；成功拯救出rPDCoV-NS6、rTGEV-ORF3，實現減毒疫苗和分子標記疫苗毒株的快速研制。Addressing challenges such as the difficulty in screening candidate strains during the traditional vaccine development process,a reverse genetics manipulation technique for porci

82、ne enteric coronaviruses has been established using yeast transformation coupled with recombination technology.rPDCoV-NS6 and rTGEV-ORF3 has enabled the rapid development of attenuated vaccines and molecularly tagged vaccine strains.四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques fo

83、r Emerging Porcine Viral Diarrhea一帶一路國際生豬產業大會3.抗豬腹瀉病毒藥物研制小分子治療藥物3.Development of Drugs Against Porcine Diarrheal Viruses-Small-Molecule Therapeuticsp針對獸用抗腹瀉病毒藥物研發難度、周期長、藥物匱乏等問題，利用人工智能技術和藥物高通量篩選技術，建立了抗腹瀉病毒的藥物高效研發平臺，篩選到芹黃素等中藥單體、硝唑尼特等小分子化合物32種。Addressing the challenges of difficulty,long development cy

84、cles,and scarcity of antiviral drugs for porcine diarrhea,we have established an efficient drug development platform for anti-diarrheal viruses utilizing artificial intelligence and high-throughput drug screening technologies.Through this platform,we have screened 32 small-molecule compounds,includi

85、ng celery flavonoids and other traditional Chinese medicine monomers,as well as nitazoxanide,that show potential against diarrheal viruses.Shi et al.,Int.Immunopharmacol,2024Zhang et al.,Virus Res,2020 Zhang et al.,Virus Res,2020姚姚晨晨等等，畜畜牧牧獸獸醫醫學學報報，2022；Yao Chen et al.,Chinese Journal of Animal and

86、Veterinary Sciences,2022;劉劉賀賀娟娟等等，畜畜牧牧獸獸醫醫學學報報，2022 Liu Hejuan et al.,Chinese Journal of Animal and Veterinary Sciences,2022.Zhang et al.,J.Agric.Food.Chem,2024四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea一帶一路國際生豬產業大會3.抗豬腹瀉病毒藥物研制飼料添加劑3.Developme

87、nt of Drugs Against Porcine Diarrheal Viruses-Feed Additives四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrheap飼糧中Se-Met的添加可提高PDCoV感染仔豬的生產性能、表觀消化率、消化酶活性和抗氧能力，緩解 PDCoV 感染仔豬引起的腹瀉，降低 PDCoV 在仔豬體內的復制。The addition of Se-Met to the diet can improve the prod

88、uction performance,apparent digestibility,digestive enzyme activity,and antioxidant capacity of piglets infected with PDCoV.It can also alleviate diarrhea caused by PDCoV infection in piglets and reduce the replication of PDCoV in their bodies.Unpublished data 未未公公布布數數據據 一帶一路國際生豬產業大會4.調控仔豬腸道健康的微生態制劑

89、研制4.Development of Microecological Agents for Regulating the Intestinal Health of Piglets病毒感染Viruses Infection腸道菌群The intestinal flora四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea腸道微生物能夠調控腸道黏膜免疫The gut microbiota can regulate intestinal mucosal

90、immunity.腸道微生物與豬腹瀉病毒感染的關系如何？How does the gut microbiota interact with porcine diarrheal virus infections?一帶一路國際生豬產業大會臨床病例分析發現腹瀉和非腹瀉豬的腸道菌群結構差異明顯Clinical case analysis has revealed significant differences in the gut microbiota composition between pigs with diarrhea and those without.PDCoV、PEDV等感染豬后，與非

91、腹瀉豬只相比，腹瀉豬的腸道菌群多樣性降低，致病菌（大腸桿菌、志賀氏菌、腸球菌等）豐度增加，有益菌豐度降低。After pigs are infected with PDCoV,PEDV,etc.,compared to non-diarrheic pigs,the gut microbiota diversity of diarrheic pigs decreases,the abundance of pathogenic bacteria(such as Escherichia coli,Shigella,and Enterococcus)increases,and the abundanc

92、e of beneficial bacteria decreases.四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral DiarrheaHuang,et al.,Front Microbiol,2019;Li et al.,Bioscience Rep,2019一帶一路國際生豬產業大會四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Vira

93、l DiarrheaZhang et al.,msystem,2024PEDV、PDCoV、PAstV、PEDV/PDCoV感染引起仔豬腸道菌群失衡，有益菌減少，致病菌增多，腸道黏膜屏障受損，仔豬發生腹瀉；PEDV,PDCoV,PAstV,and PEDV/PDCoV infections can cause an imbalance in the gut microbiota of piglets,leading to a decrease in beneficial bacteria,an increase in pathogenic bacteria,damage to the inte

94、stinal mucosal barrier,and diarrhea in piglets.PDCoV強、弱毒株感染仔豬后，強毒株感染引起仔豬腸道菌群失衡，腸道黏膜屏障受損，仔豬發生腹瀉。After piglets are infected with virulent and attenuated strains of PDCoV,the virulent strain can cause an imbalance in the gut microbiota and damage to the intestinal mucosal barrier,resulting in diarrhea

95、in piglets.Li et al.,Microbiologyopen.2020;Shu et al.,Virus Res.2022腹瀉病毒感染影響仔豬腸道菌群的穩定性Diarrheal virus infections can affect the stability of the gut microbiota in piglets.一帶一路國際生豬產業大會四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea隨著豬的日齡增大，腸道菌群多樣性增

96、加，定植在腸道中的益生菌就越多，豬的腹瀉程度也越輕；As the age of pigs increases,the diversity of the gut microbiota also increases,leading to a greater number of probiotics colonizing the intestine and subsequently lighter diarrhea symptoms in pigs.豬的日齡越小，腹瀉越嚴重，腸道菌群多樣性降低，有害菌豐度升高。Conversely,younger pigs tend to experience mo

97、re severe diarrhea,which is associated with a decrease in gut microbiota diversity and an increase in the abundance of harmful bacteria.Unpublished data未未公公布布數數據據不同日齡豬感染PDCoV其腸道菌群組成與臨床癥狀相關The composition of the gut microbiota in pigs infected with PDCoV at different ages is related to their clinical

98、 symptoms.一帶一路國際生豬產業大會四、新發豬病毒性腹瀉防控技術研究IV.Research on Prevention and Control Techniques for Emerging Porcine Viral Diarrhea建立仔豬糞菌移植模型，成年豬糞便菌群在仔豬腸道的定植可改變仔豬腸道菌群的結構組成，維持仔豬腸道健康，降低腸道炎癥反應，緩解PDCoV感染引起的腹瀉癥狀。By establishing a fecal microbiota transplantation model for piglets,the colonization of the adult pig

99、s fecal microbiota in the piglets intestine can alter the structural composition of the piglets gut microbiota,maintain intestinal health,reduce intestinal inflammatory responses,and alleviate diarrhea symptoms caused by PDCoV infection.Unpublished data未未公公布布數數據據通過調控仔豬腸道菌群可阻遏PDCoV對仔豬的感染The infection

100、 of piglets with PDCoV can be blocked by regulating their gut microbiota.一帶一路國際生豬產業大會致 謝Acknowledgments感謝“一帶一路”國際生豬產業大會組委會、國家自然科學基金委、河南農業大學的支持，動物分子病原學團隊成員的付出！We are grateful for the support provided by the Organizing Committee of the Belt and Road International Pig Industry Conference,the National Natural Science Foundation of China,and Henan Agricultural University.Special thanks to the members of our Animal Molecular Pathogenology team for their dedication and efforts!一帶一路國際生豬產業大會敬請批評指正！謝謝！Thank you!一一帶帶一一路路國國際際生生豬豬產產業業大大會會Belt and Road International Swine Industry Conference一帶一路國際生豬產業大會