猪病毒性腹泻科学防控 Prevention and Control of Porcine Viral Diarrhea 冯力 Li Feng 中国农业科学院哈尔滨兽医研究所 Harbin Veterinary Research Institute (HVRI), Chinese Academy of

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1 猪病毒性腹泻科学防控 Prevention and Control of Porcine Viral Diarrhea 冯力 Li Feng 中国农业科学院哈尔滨兽医研究所 Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences (CAAS)

2 p 猪病毒性腹泻基本特点与危害 Characteristics and cost of porcine viral diarrhea p 猪病毒性腹泻科学防控要点 Key points for prevention and control of porcine viral diarrhea

3 p 猪病毒性腹泻基本特点与危害 Characteristics and cost of porcine viral diarrhea

4 引起腹泻的因素 Causes of Diarrhea ( 猪只 ) 腹泻 ( 下痢, 拉稀 ) 是很多疾病发生时的一种临床表现 ( 症状 )(Pigs) diarrhea (dysentery, diarrhea) is a clinical sign (symptom) of many diseases. 病因 Causes 传染性因素 Infectious factors 非传染性因素 Non-infectious factors 细菌 Bacteria 病毒 Virus 寄生虫 Parasites 蛇形螺旋体 Serpentine spirochetes 疫病 Disease 病原 Pathogen 病原分类 Types of Pathogen 血清型 Serotype 猪传染性胃肠炎 TGE 猪传染性胃肠炎病毒 TGEV 冠状病毒 Coronavirus 1 猪流行性腹泻 PED 猪流行性腹泻病毒 PEDV 冠状病毒 Coronavirus 1 猪轮状病毒感染 PoRV 猪轮状病毒 PoRV 呼肠孤病毒 Reovirus 多血清型 Multiple serotypes

5 福建某猪场发病情况 Incidence of a farm in Fujian

6 2014 年东北某猪场出现的腹泻 Diarrhea occurred in a Northeast China farm in 2014 成猪腹泻 ( 母猪与种公猪 ) Adult pigs diarrhea (Sows and Boars)

7 2015 年 8 月广东高州 August 2015 Gaozhou, Guangdong

8 猪病毒性腹泻的特点 Characteristics of porcine viral diarrhea p 呕吐, 水样腹泻, 脱水, 哺乳仔猪的高死亡率 Vomiting, watery diarrhea, dehydration, and high suckling pig mortality p 无体温升高, 濒死期体温降低 No increase in body temperature, low body temperature near death p 所有日龄猪只均可感染, 日龄增加, 死亡率下降 found in all ages, mortality declines in old pigs p 病原为病毒, 具有极强的传染性 Pathogen is virus, high infectious p 冬春季节多发, 夏季发生也成新常态 Usually occurs in spring and winter, but summer break becomes new normal p 抗菌素治疗无效 Antibiotic shows no effect

9 危害与损失 damage and loss p 仔猪死亡 (7 日龄仔猪的高死亡率可高达 100%, 每头仔猪的直接饲料成本约 200 元 )Piglet Death (100% mortality for 7- day piglets, direct feed cost is about 200 per piglet) p 育肥猪推迟出栏 ( 推迟出栏 10~15 天左右, 育肥后期每天需要 3kg 饲料,1.5 元 /kg, 每头损失 45 元 )Delayed slaughtering (10-15 days, 3kg feed/day in late finisher, 1.5 / kg, lose 45 per head) p 2013 年我国南方某 5000 头能繁母猪场, 由腹泻损失仔猪达到头 (2 次 )2013, a 5000-sow farm in southern China, the loss of piglets caused by diarrhea was 15,629 (twice) 美国 2013 年猪流行性腹泻损失仔猪 600 余万头 piglet loss caused by PED in the US was > 6 million in 2013

10 p 猪病毒性腹泻科学防控要点 Key points for prevention and control of porcine viral diarrhea

11 明确病因 Determining the causes 免疫预防 Vaccination 生物安全 Biosecurity

12 明确病因 ---- 实验室诊断, 确诊 Determining the causes - laboratory diagnosis, confirmation A C E B D F A,B:TGE 传染性胃肠炎 C,D:PED 猪流行性腹泻 E,F:PRV 伪狂犬

病毒性腹泻病的实验室诊断方法 Diagnosis methods for porcine viral diarrhea (1) 血清学诊断

immunofluorescence 间接 ELISA Indirect ELISA (2) 病原学诊断 Pathogenic diagnosis

病毒的核酸检测 Virus nucleic acid testing : RT-PCR 核酸探针 RT-PCR Nucleic acid probe

13 病毒性腹泻病的实验室诊断方法 Diagnosis methods for porcine viral diarrhea (1) 血清学诊断 Serological diagnosis 中和试验 (VN) Virus Neutralizing (VN) 间接免疫荧光 Indirect immunofluorescence 间接 ELISA Indirect ELISA (2) 病原学诊断 Pathogenic diagnosis 病毒的分离与鉴定 Virus Isolation and identification 电镜负染 EM negative staining 病毒的核酸检测 Virus nucleic acid testing : RT-PCR 核酸探针 RT-PCR Nucleic acid probe 直接免疫荧光 Direct immunofluorescence 双抗体夹心 ELISA Sandwich ELISA 免疫胶体金 Immune colloidal gold

14 免疫胶体金检测 TGE PED 和 PRV Detecting TGE,PED and PRV by Immune colloidal gold A C D B A:PED 阴性 Negative B:PED 阳性 positive E:PRV 阳性 positive C:TGE 阴性 Negative D:TGE 阳性 positive F:PRV 阴性 Negative E F

15 中国猪腹泻病原的复杂性 Complexity of pathogen for porcine diarrhea in China Sample detecting rate Farm detecting rate

16 Detection of porcine delta coronavirus in China 猪冠状病毒在中国的检测 PDCV PCR Porcine delta coronavirus is detected in 5 provinces in China, the positive rate of PDCV is about13.99% (20/143). 中国 5 省都检测到 delta- 冠状病毒阳性

17 免疫原的种类 Immunogenic types p 强毒免疫 wild virus vaccination p 自家疫苗 in-house vaccine p 非正规疫苗 ( 白瓶苗, 走私苗 ) Unlicensed vaccines(white bottle vaccines, smuggled vaccines ) p 灭活疫苗 Inactivated vaccine p 活疫苗 ( 单苗多联苗 )Live vaccines (single vaccines, multi-valent vaccines)

猪流行性腹泻疫苗的研究 Research on PED Vaccines China Japan South Korea CV777 p125 P-5V KPED-9 DR13 PED Vaccines are developed in China 中国 PED 疫苗研发 p Bi-combined killed vaccine against TGEV and PEDV(1996)

18 猪流行性腹泻疫苗的研究 Research on PED Vaccines China Japan South Korea CV777 p125 P-5V KPED-9 DR13 PED Vaccines are developed in China 中国 PED 疫苗研发 p Bi-combined killed vaccine against TGEV and PEDV(1996) TGEV/PEDV 二联灭活苗, 1996 pbi-combined live vaccine against TGEV and PEDV(2003) TGEV/PEDV 二联活苗,2003 ptri-combined live vaccine against TGEV, PEDV and PoRV (G5) (under way) TGEV/PEDV/PoRV 三联活苗 ( 进行 )

19 美国猪流行性腹泻疫苗的研究 U.S. Research on PED Vaccines 猪流行性腹泻灭活疫苗 PED inactivated Vaccine

20 美国农业部批准的 PED 疫苗, RNA

21 疫苗的科学免疫 Scientific immunization p 做到无易感猪群 ( 群防 ); 如果育肥猪先发的猪场, 更应该引起重视 Ensure no susceptible herd (whole-herd prevention); if break first in finishing pigs, should pay more attention p 哺乳仔猪靠被动免疫, 要求做好母猪, 尤其是后备母猪免疫是重点 Suckling pigs depend on passive immunization, we should focus on sow vaccination, especially gilts vaccination. p 抗体对活疫苗有干扰, 要做到正确的免疫 The antibody interferes with live vaccines, ensure to have the correct vaccination

22 谁是最易感猪群?Which one is the most susceptible herd? p 后备母猪 Gilts p 后备公猪 Boars p 断奶保育与育肥 Weaning, nursery, and finishing pigs

23 疫苗的科学免疫 Scientific use of vaccine p 疫苗不是用来治疗用的, 要牢牢树立传染病根本在于防 The vaccine is not for treatment. Prevention is the key. p 腹泻的季节性 ( 冬春季节 ) 已经不明显, 做好日常免疫, 尤其是稳定期间的基础免疫非常关键 Seasonality of diarrhea (spring and winter) becomes less noticeable. Routine immunization, especially basic immunization during stable period is critical. p 反复反饲 ( 强毒免疫 ) 导致猪群中长期大量的野毒存在, 可能让我们的防控更加困难 Repeated feedback (virulent immunity) leads to a large amount of wild viruses in the herd that makes prevention and control more difficult.

24 疫苗的科学免疫 Scientific use of vaccine p 先活苗, 后死苗 live vaccine first, then killed vaccine p 先死苗, 活苗后 killed vaccine first, then live vaccine p 感染或反饲 = 活苗免疫 Infection or feedback = live vaccine

25 群体中易感动物存在 Susceptible animals in herd are present 传染源 (Source of infection) 传播途径 (Route of transmission) 易感动物 (Susceptible animals) 强毒 (virulent) 强毒 (virulent) 强毒 (virulent) 做好群体免疫, 消灭易感猪群 (do herd vaccination and eliminate susceptible pigs) 强毒感染 (Infected with virulent strains)

26 免疫接种 (Immunization) 2012 河北某场腹泻发生情况 Porcine diarrhea in a farm in Hebei, 2012 前几年的腹泻情况 (Diarrhea several years ago) 母猪 (sow) 免疫接种 (Immunization) 年腹泻情况 (Diarrhea in ) 母猪 (sow) 开始 (begin) 育成哺乳仔猪育成哺乳仔猪 (finishing pig) (suckling pig) (finishing pig) (suckling pig) 保育猪断奶仔猪保育猪断奶仔猪 (growing pig) (weaned pig) (growing pig) (weaned pig) 母猪 (sow) 发病 (sick) 育成猪 (finishing pig) 哺乳仔猪 (suckling pig) 不发病 (healthy) 后来 (later) 保育猪断奶仔猪 (growing pig) (weaned pig)

27 免疫保护与强毒感染的关系 The relationship between immune protection and virulent infection 强毒的量体液免疫与细胞免疫 (Dose of virulent virus) (Humoral immunity and cellular immunity) 免疫失败 (Immunity failure)

28 科学的生物安全措施 Scientific biosecurity measures p 彻底的清洗消毒, 空舍 (7-14 天 )Thorough cleaning and disinfection, empty rooms (7-14 days) p 真正的全进全出, 多点饲养 True all in/out, multisite production 缓冲 p 尽可能将后备放到一栋舍 buffer Try to put all gilts in one building p 冬季产子舍保温 Keep farrowing rooms warm in winter p 坚持自繁自养 have both multiplication and production p 避免外来人员车辆等进入生产区 Avoid outside personnel and vehicles entering the production area 门 Door 走廊 Corridor 猪舍 Rooms 单元 1 Unit1 单元 2 Unit 2 单元 3 Unit 3 单元 4 Unit 4 单元 5 Unit 5

29 冲洗干净, 空舍 7 天 Rinse it, empty rooms for 7 days 戊二醛消毒 : 当 ph 值为 7.5~8.5 时, 作用最强 For glutaraldehyde: when ph is between 7.5~8.5, it works the best s 推荐一种消毒方法 Recommend a disinfection method 物理清洗, 再用杜邦公司的传清浸泡 2 小时, 冲洗 Physical cleaning, soak 2-hour with DuPont Disinfectants, and then wash 用有机氯消毒, 空 2 天 sterilize using organic chlorine, empty rooms for 2 days 戊二醛消毒, 空 2 天,sterilize using glutaraldehyde, empty rooms for 2 days

30 病毒性腹泻病原的传染方式 Transmission of viral diarrhea pathogen 直接途径 : 粪口途径, 消化道接触污染物 Direct path: fecal-oral contact, digestive tract contacts with contaminated materials 间接途径 : 车辆饲料车, 运猪车 - 服务人员装备, 包括饲料在内的其他污染物 Indirect path: Vehicles - feed truck, Pig transportation trucks service staff equipment, including other contaminated materials like feed

31 病原在体外的存活时间 Survival time of pathogen in vitro p 病毒的体外存活时间与温度与湿度有关 Survival time in vitro is related to temperature and humidity p 4 C 条件下, 在泥浆中至少存活 28 天 At 4 C, it can survive in the mud for at least 28 days. p 25 C 条件下, 在粪便污染的干饲料中存活 7 天 At 25 C, it can survive in the dry feed contaminated by feces for 7days. p25 C 条件下, 在湿的饲料中存活 14 天 At 25 C, it can survive in the wet feed for 14 days. p25 C 条件下, 在湿的饲料混合物种至少存活 28 天 At 25 C, it can survive in the wet feed blend for at least 28 days.

32 理化特性 Physicochemical properties p 大于 60 C 病毒失去活性 When > 60 C, virus is inactivated p 在 37 C,pH 条件下病毒稳定 When at 37 C,pH , virus is stable p 4 C,pH 5-9 条件下稳定 When at 4 C,pH 5-9, virus becomes stable

33 敏感性 Sensitivity 1% 福尔马林 1% formalin 无水碳酸纳 Anhydrous sodium carbonate 脂溶性溶剂 Lipid-soluble solvent 溶于磷酸的 1% 离子去污剂 1% phosphoric acid dissolved ionic detergents 2% 氢氧化钠 2% sodium hydroxide

34 明确病因 Determining the causes 免疫预防 Vaccination and Prevention 生物安全 Biosecurity

35 控制传染性腹泻发生的关键 Keys for controlling the incidence of Infectious diarrhea 引种 Introducing genetic 发病猪 Sick pigs 带毒猪 Infected pigs 环境 Environment 人员 Personnel 工具 Tools 粪口途径 fecal-oral contact 接触病原及污染物 contact with Pathogen & contaminated materials 非免疫猪群 Non-vaccine herd 抗体低下猪群 Low immunity herd 免疫失败猪群 vaccination failure herd 传染源 Source of infection 传播途径 Transmission routes 易感动物 Susceptible animals 免疫 Immunity 生物安全 Biosecurity 管理 Management 消毒, 空舍 Disinfection, empty rooms 疫苗免疫 Vaccination 接触 Contact 隔离 quarantine 疫苗免疫 Vaccination 群防 whole-herd prevention

36 克服病毒性腹泻疾病的三个误区 Three Misunderstandings in fighting viral diarrhea diseases u 第一个是诊断误区 The first is diagnosis p 诊断想当然, 仅凭借临床症状来判断 diagnose based on clinical sign and experience, not on data p 一次实验室诊断结果用几次或一两年 use one-time diagnostic result for several times or one or two years p 实验室诊断不全面 diagnosis is incomplete p 试验室检测水平不同, 影响结果的科学与真实 variability in laboratory testing influences fact seeking

37 u 第二个是免疫误区 The second is vaccination p 不当传染病对待, 只是局部免疫 ( 母猪免 )do not treat it as infectious diseases, only do partial vaccination (sows) p 经产与后备母猪一样免疫 Same vaccination for P2+ and gilts p 等到发病了, 才接种疫苗, 疫苗当治疗制剂 Only vaccination when break, using vaccination as therapeutic treatment p 只有流行季节前做, 平时基本不进行免疫 Only vaccination in epidemic seasons, not vaccinate routinely p 过度依赖疫苗, 不重视生物安全 Rely on vaccines excessively, care less about biosecurity p 反复返饲 Repeatedly feedback

38 克服病毒性腹泻疾病的三个误区 Three misunderstandings in fighting viral diarrhea diseases u 第三个是生物安全误区 The third is in Biosecurity p 做不到真正的全进全出 Incomplete all in/ out p 消毒不彻底, 空舍时间过短 Incomplete sterilization, room empty time not long enough p 不把引种隔离当回事 Don t pay attention to introducing live animals and quarantine p 综合管理出现漏洞 Loopholes in integrated management

39 防控的关键点 Keys to control and prevention p 确定发病病原! 确诊 Determine the pathogen! Confirmation p 做好全群免疫, 不留易感猪群 ( 尤其是做好后备母猪初产母猪, 后备公猪的免疫 )!Do whole-herd vaccination, leaving no susceptible pigs behind! (especially for gilts and boars) p 列入常规免疫计划而非季节性免疫 include it as routine vaccination plan instead of seasonal vaccination p 严格的生物安全措施 : 多点饲养, 全进全出, 严格的消毒与空舍!Strict biosecurity measures: multi-site production, all in/ all out, strict disinfection and empty rooms! p 做好产子舍保温 keep farrowing rooms warm p 避免应激, 提高机体自身抗病力 Avoid stress, increase the body's own resistance to disease

40 Thank you for your attention! 谢谢! Dr. LI FENG Harbin Veterinary Research Institute (HVRI) Chinese Academy of Agricultural Sciences (CAAS) mobile: Fax:

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