Use of Histopathology to Solve Complex Diagnostic Problems in Pig Production 利用组织病理学解决养猪生产系统中 复杂的诊断问题 James E. Collins, DVM, PhD, Diplomate ACVP Professor and Director Minnesota Veterinary Diagnostic Laboratory 詹姆斯 考林斯博士教授 & 主管美国明尼苏达大学, 兽医诊断实验室
Case No. 1 Diarrhea 案例 1- 腹泻 High Morbity and Mortality 高发病率和死亡率
Source: Dr. Ackerman 来源 : 阿克曼博士
Source: AASV
Source: Dr. Ackerman 来源 : 阿克曼博士
Diagnostics 诊断 Same basic guidelines of diarrhea diagnostic workups 与腹泻诊断相同的基本准则 Select live, acutely affected pigs with watery diarrhea and vomiting 选择伴有腹泻和呕吐的急性感染活猪 Perform necropsy and collect samples within 15 minutes of euthanasia 在安乐死的 15 分钟内进行剖检和样本采集 Gross lesions: liquid intestinal contents 肉眼观察病变 : 肠内液体容物
Sample collection 样品采集 Small intestine: 小肠 : 6-10 sections representing the whole length of the small intestine 6-10 段代表小肠的整个长度 10 cm segments fresh and 2 cm segments fixed 10 厘米新鲜样本和 2 厘米固定样本 Other tissues for a complete workup and to rule out differential diagnoses 采集其它组织来进行完全诊断, 排除差异诊断 Stomach, colon, tonsil, lymph node, lung, heart, spleen, liver, kidney, brain -- fresh and fixed 胃, 结肠, 扁桃体, 淋巴结, 肺, 心脏, 脾, 肝, 肾, 脑 - 新鲜和固定样本
TGE/PED Newspaper gut 猪传染性胃肠炎 / 猪流行性腹泻 报纸肠 (you could read a newspaper through it) 可以透过它读报纸
Necropsy 剖检 Virus isolation: 病毒分离 Enteric viruses - difficult to isolate in vitro 肠道病毒 - 难以在体外分离 Electron Microscopy: 电镜 low sensitivity 低灵敏度 Histopathology: 组织病理 : atrophic enteritis 萎缩性肠炎 same as TGEV and Rotavirus 与猪传染性胃肠炎和轮状病毒相同 Real time PCR: 实时 PCR sensitive and specific 敏感和特异 available in US laboratories 美国实验室提供
PEDV diagnostic tests 猪流行性腹泻病毒诊断检测 IHC, ISH: less sensitive than PCR 比起 PCR 较不敏感 good for research, retrospective studies 对科研, 回顾性研究有益 ELISA: Biovet in the US & Canada Biovet 在美国和加拿大
Porcine Epidemic Diarrhea 猪流行性腹泻 Real-time PCR assay for detection and differentiation of PEDV and TGEV in clinical and environmental samples. 实时荧光定量 PCR 检测能在临床和环境样本中检测到和区分猪流行性腹泻病毒和感染性肠胃炎病毒 This test can be performed on different types of samples. 这种测试可以用不同类型的样品 PCR test three-in-one: TGEV + PEDV + SDCV like duplex PEDV/TGEV it detects and differentiate viruses of PEDV and TGEV and also Swine delta coronavirus (SDCV). PCR 检测三合一 : 猪传染性胃肠炎 + 猪流行性腹泻病毒 + delta- 轮状病毒, 像复式 PEDV/TGEV 一样, 它能检测和区分猪流行性腹泻病毒和猪传染性胃肠炎病毒和 delta- 猪轮状病毒 ELISA test allowing to detect IgG antibodies to the PEDV nucleoprotein is now available. 能够检测到针对 PED 病毒核蛋白的 IgG 抗体的 ELISA 测试现在已可用 Serological test (IFA) for detection of antibodies against the virus in serum samples. 血清学试验 (IFA) 能够检测血清样品中抗病毒的抗体 Sampling kit and protocol to help you perform environmental sampling 采样试剂盒和流程能够帮助你进行环境采样
Porcine Epidemic Diarrhea 猪流行性腹泻 PEDV replicates in the epithelial cells of the small intestine PED 病毒在小肠的上皮细胞进行复制 Incubation period: 1-2 days 潜伏期 : 1-2 天 Shedding in feces: 7-11 days 从粪便散毒 :7-11 天 Immune response: mucosal and serological: serum antibodies at 7-14 dpi 免疫应答 : 粘膜和血清学 : 在攻毒 7-14 后的血清抗体 Source: Kim et al. 1999
Porcine Epidemic Diarrhea 猪流行 性腹泻 PED is caused by a coronavirus (PEDV) 猪流行性腹泻是由冠状病毒引起的 (PEDV) PEDV causes diarrhea and vomiting in swine of all ages PED 病毒能引起所有年龄猪腹泻和呕吐 High mortality in piglets under 2 weeks of age 在低于 2 周龄仔猪中有高死亡率
Porcine Epidemic Diarrhea 猪流行 性腹泻 PEDV first described in Europe in the 70s 第一宗猪流行性腹泻病毒是 70 年代在欧洲被确认 PED is an important problem in USA & Asia 猪流行性腹泻在美国和亚洲都是一个严重的问题
E. Coli Edema disease 大肠杆菌水肿病
Edema Disease pig Stomach 水肿病猪的胃
E. coli toxin induced vascular necrosis 大肠杆菌毒素导致的血管坏死
Other sample types 其他样本类型 Feces (5 ml) 粪便 (5 毫升 ) Fecal swabs 粪便拭子 Feedback material 返饲材料 Environmental swabs 环境拭子 Feed (under development) 饲料 ( 开发中 )
Case No. 2 案例 2 Bloody diarrhea and increased mortality in gilts in isolation barn 隔离后备母猪血性腹泻和死亡率的增加 Gilts recently arrived after 8 hour truck ride 经过 8 小时运输刚到的后备
Parasites 寄生虫 Trichuris suis 猪鞭虫 Large intestine, ulceration, necrosis 大肠, 溃疡, 坏死
Brachyspira pilisicoli 短螺旋体 pilisicoli Bacteriology weakly beta-hemolytic, PCR typing on isolates 细菌学 - 弱乙型溶血性, 对菌株的 PCR 分型
Telepathology 远程病理学 Gross & microscopic pathology at a distance 远程组织和显微病理学 Real-time dynamic systems 实时动态系统 Static (eg. whole slide scanning) 静态 ( 例如整个切片扫描 ) 21 st century approach to veterinary pathology 21 世纪兽医病理学的方法 collaborative international training 国际合作培训 real-time professional consultation 实时专业咨询 public private partnerships 公私合作伙伴关系
IVCC / Zoetis Grant IVCC/ 硕腾资助 Year 1 telepathology partnership objectives 第 1 年远程病理学合作的目标 1) Establish telepathology equipment 建立远程病理学设备 UMN & Jilin University in China 美国明尼苏达学和中国吉林大学 2) Develop online training modules 发展网上培训课程 Use of equipment and real-time collaboration 使用设备和实时协作 3) Collaborative pathology training modules 联合病理学培训课程
Equipment for Telepathology 远程病理学的设备 Grossing pathology system 组织病理学系统 Microscope Telepathology unit 显微远程病理学系统
Telepathology addressing needs across all 3 missions 远程病理学通过三方面解决需要 Education 教育 DVM students 兽医学院学生 CE 继续教育 Training 培训 Research 科研 Service 服务 International Telepathology Partnership to Improve Pathology Training for Swine Diagnostics in China IVCC 2014-15 partially support the initiation of this program 国际远程病理学在中国旨在提高猪诊断病理培训合作项目 -IVCC 2014-15 部分支持这一计划的启动
Current uses and expansion 目前的用途和扩展 Collaboration 协作 Swine pathology & diagnostic consultation with Chinese partners 与中国合作伙伴开展猪病理学和诊断咨询 Teaching 教学 Pathology cases 病理案例 UMN s Cancer Center 明尼苏达大学癌症中心 Veterinary-Human pathology collaboration 兽医 - 人体病理学合作
Successfully established international telepathology partnership! 成功建立了 国际远程病理学的合作伙伴!