PowerPoint Presentation

Size: px

Start display at page:

Download "PowerPoint Presentation"

贡欣梁
5 years ago
Views:

1 Use of lung homogenate to acclimate gilts to Mycoplasma hyopneumoniae 利用肺匀浆驯化后备母猪抵抗肺炎支原体 Luc Dufresne DVM 卢克迪弗雷纳, 兽医博士 Rebecca Robbins DVM, PhD 丽贝卡罗宾斯, 兽医博士, 博士 Seaboard Foods

2 Mhp and the breeding herd 肺炎支原体与繁殖群 Prevalence of Mhp positive pigs at weaning correlated with Mhp pneumonia Mhp 阳性断奶仔猪的发病率与 Mhp 肺炎有关 Colonization of piglets with Mhp is associated with presence of positive lactating sows 仔猪感染 Mhp 与阳性哺乳母猪有关 Vaccination doesn t prevent Mhp colonization, alter Mhp transmission or consistently reduce Mhp shedding 疫苗接种不能预防 Mhp 的定植, 改变传播以及持续降低 Mhp 的排毒量

3 Mhp and the breeding herd, cont d 肺炎支原体与繁殖群 ( 接上页 ) Success of elimination is inconsistent 成功消除的结果不一致的 3.1 to 20% failure rate 3.1% 到 20% 的失败率 Introduction of infected genetic stock and aerosol transmission 引入感染的品种和气溶胶传播 Entry of naïve gilts into a positive herd or gilts exposed prior to entry are the source of ongoing Mhp infection 阴性后备母猪进入阳性群或后备进群前感染是 Mhp 感染的来源

4 腔液阳性百分比周龄口Time of infection highly variable within and between GDUs of an integrated system 在综合生产系统体系后备培育舍内部和之间的感染事件差异变化明显 Longitudinal monitoring of Mhp in 4 different GDU flows of a large integrated system. Figure from Anderson, et al 2016.

5 Observations we made 我们观察到的结果 Growing gilt population no longer Mhp PCR positive 生长的后备母猪群不再有 Mhp PCR 阳性 Gilts were coughing, thumping and oral fluids tested positive for Mhp within 2-4 weeks after entering sow farm 后备母猪在进入母猪场后 2-4 周内, 出现咳嗽撞栏的现象, 同时口腔液体检测其为 Mhp 阳性 Gilts would either not come in to heat or return following 1 st breeding event. 后备或者不发情或第一次配种后返回 Gilts that returned generally came back in to heat and successfully conceived after the 2 nd service. 一般来说, 后备返回后发情, 第二次配种后成功受孕 Mhp was the only consistent diagnosis we had across cells, pods and farms. Mhp 是我们在栏位猪舍和农场唯一的结果一致的诊断

6 Interventions we have tried to control Mhp none are long term solutions 我们尝试的用来控制 Mhp 的干预方法, 没有一种是长期有效的方案 Aivlosin in water at 2.5 mg/kg for 7 days beginning 2 weeks prior to arrival (gilts only) 在入场前 2 周连续 7 天在水中按 2.5mg/kg 的剂量艾罗西宁 ( 只给后备母猪 ) Lincomycin 100 g/ton from time of breeding until 2 weeks post-breeding (gilts only) 从配种到配后 2 周按 100g/ 吨的剂量给予林可霉素 ( 只给后备母猪 ) Lincomycin 100 g/ton in 2 week pulses over 6 week period (all females in B&G) 每 2 周内给予 100g/ 吨的林可霉素持续 6 周时间 ( 所有配种和妊娠母猪 ) CTC 1200 g/ton 2 weeks and Lincomycin 100 g/ton 3 weeks for 7 week period (all females in B&G) 两周一次按 1200g/ 吨的剂量给予 CTC 同时配每 3 周一次的林可霉素, 共持续给药 7 周 ( 所有配种和妊娠母猪 ) Draxxin or Lincomycin 5 mg/lb administered 1x at placement (gilts only) 给予 1x 的瑞可新或林可霉素 5mg/ 磅 ( 只给后备母猪 ) Vaccinate gilts with RespiSure at processing, 3, 6, and 16 or 22 weeks of age. 在后备母猪第 3,6,16 或 22 周时接种 RespiSure 进行免疫

7 So it was time to try something new 是时候尝试新的方法了

8 Goals for Seaboard Mhp Acclimation using planned exposure 西宝德进行 Mhp 驯化的计划感染的目标 Prevent Mhp associated disease in growing/finishing pigs 预防成长及育肥猪的与 Mhp 有关的疾病 Improve reproductive performance of gilts once entered into Mhp positive sow farms 提高进入 Mhp 阳性母猪场的后备母猪的繁殖能力 While avoiding overlap of concurrent acclimation processes that could negatively affect health or performance of growing gilt 避免驯化过程的重叠, 这可能会对后备母猪的健康或生长能力产生负面影响

9 Goals 目标 Part 1. 第 1 部分 Make a lung tissue homogenate* containing Mhp to initiate Mhp colonization of growing gilts 制备含有 Mhp 的肺组织匀浆以免疫生长后备母猪 *Note: exposure to live organisms is a widely practiced method to promote seroconversion to microorganisms when there is no commercial vaccine available or strain-specific immunity is desired 注意 : 当没有商业疫苗可用或需要的疫苗株产生特异性免疫时, 应用新鲜组织进行免疫是广泛应用的方法以促使动物对微生物的血清转阳

10 Working seed 制备有效免疫源 Make master seed from lung of clinical gilt 从临床后备的肺制备免原源 31 wks old; Mhp Ct 25.62; PRRS, SIV, Mhr, Pasteurella Neg 31 周龄, Mhp Ct 值 25.62; 蓝耳流 Mhp 阴性 Intra-tracheal inoculation as previously described 和之前描述的一样, 采用气管内接种 Administered 1 time 接种一次 Ceftiofur administered 3 days after inoculation 接种后 3 天注射头孢塞弗 Laryngeal tracheal swabs collected before harvest 收集喉部气管拭子 Test for Mhp by PCR 通过 PCR 检测 Mhp Bronchial swabs collected before homogenizing 在采取肺匀浆前收集支气管拭子 Test for Mhp by PCR 通过 PCR 检测 Mhp We observed bronchial swabs avg 7 Cts lower than laryngeal swabs (equivalent to 2 logs greater Mhp CCU). 我们观察到气管拭子平均值比喉部拭子低 7 Cts( 相当于高于 Mhp CCU 2 个 logs)

11 Working seed 制备有效免疫源 Homogenize tissue 70:30 with medicated Friis media (ISU VDL, Ames, IA) 用 Friis (ISU VDL, Ames, IA) 与培养的匀浆组织按 70:30 混合 Previously used only lesioned tissue 之前仅使用病变组织 Now using ENTIRE lung 现在使用全肺 QC for Working Seed (or what we have found that worked for us) 有效的免疫源的质量控制 Mhp* Mhr PRRS SIV PCV2 Pasteurella Other Bacteria Ct <24 Ct >33 Neg Neg Pos No growth No growth 10 4 CCU/ml Ct >26 * Perform in triplicate and take the average

87 1 - Negative 2 - Ct 20.33 2 - Ct 21.35 2 - Ct 27.58 3 - Ct 21.

12 Comparison of Mhp Ct by location 各部位 Mhp Ct 值对比 Ct for Sections in Triplicate: 各部位三份病料的 Ct 值 : 1 - Ct Ct Negative 2 - Ct Ct Ct Ct Ct Ct Average Ct for Sections: 各部位平均 Ct 值 : 1 - Ct Ct Ct

13 Comparison of lung homogenates Lesioned lung only 病变肺组织 Ct-average 平均值 22.75* Ct-range 范围 16.08, 40* 肺匀浆的对比 Entire lung 全肺 Ct-average 平均值 Ct-range 范围 18.46, Yield (ml) 产量 (ml) 150 PCR was not run in triplicate PCR 未按一式三份运行 Yield (ml) 产量 (ml) 390

14 Goals 目标 Part 1. 第一部分 Make a lung tissue homogenate to initiate colonization of population 使肺匀浆开始群体定植 Part 2. 第二部分 Using working seed administered intra-tracheally colonize growing gilts with Mhp at a specified time 使用制作的 Mhp 免疫源在特定时间气管内接种至生长的后备母猪 Part 3. 第三部分 Infect growing gilt with Mhp by 15 weeks of age using naturally colonized seeders 使用 Mbp 天然免疫源在 15 周龄时给成长后备母猪免疫

Mhp Exposure Mhp 接种 Used 50-350 timeline 在 50-350 时间段内进行 Begin exposure at 42 days of age 在 42 天日龄时开始接种 To infect

15 Mhp Exposure Mhp 接种 Used timeline 在时间段内进行 Begin exposure at 42 days of age 在 42 天日龄时开始接种 To infect population by 15 weeks of age 在 15 周龄以前感染 Transmission 传播 = 0.77 (Roos, et al. 2016) Figure from Pieters and Fano, 2016

16 Materials 材料 14 FR. Straight tip male catheter 14FR 的直头导管 12 cc luer-lock syringe 12cc 的鲁尔锁注射器 Lung homogenate (1 ml per pig) 肺匀浆 ( 每头猪 1ml) Friis media (9 ml per pig) Friis 介质 ( 每头猪 9ml) Sterile mason jar (to mix homogenate with the media = 10 ml volume per pig) 无菌梅森罐 ( 把匀浆和媒介混合 = 每头猪 10ml) Laryngoscope (optional) 喉镜 ( 可选的 )

17 1 st Attempt: 第一次尝试 Establish infection using Intra-tracheal inoculation then roll naturally infected seeders 使用气管内接种建立感染, 然后生产自然感染源 Intratracheal (IT) seeder at 1:5 免疫原按 1:5 稀释气管内接种 Higher b/c of 4 week lag time to infection 4 周滞后时间较高的 b/c 感染 4 weeks later begin using Natural seeder at 1:9 4 周后开始将免疫原按 1:9 稀释进行自然感染 Roll from oldest room no sooner than 4 weeks after IT exposure 在气管内感染后的 4 周内, 从最先的房间滚动 Continue for each subsequent lot 继续每个后续批次

18 Determine if Mhp exposure occurred 确定 Mhp 接种是否有效 Longitudinal sampling of a lot 各批次进行纵向抽样 convenience sample within the lot 批次内方便样品 Oral fluids 口腔液体 10, 15, 17, 19 and 32 weeks of age 第 10,15,17,19 和 32 周进行采样 4 per lot at each time point 每个时间点每批次采集 4 个样本 Serum 血清 15, 17, 19 weeks of age 在第 15,17,19 周采集血清样本 30 per lot at each time point 每个时间点每批次采集 30 个样本

19 Mhp infection successfully established using seeder pigs 被接种的猪成功感染 Mhp 口腔液体 -IT& 天然种子的 Mhp PCR 结果阳性百分比 (%) 最小 ct *All oral fluids at 10 weeks were PCR negative. * 所有的口腔液体在第 10 周都是 PCR 阴性

20 Mhp infection successfully established using seeder pigs-serology 通过血清学方法证实接种猪成功建立 Mhp 感染 SP 阳性 % 年龄 ( 周 ) % Positive SP Age (weeks) IT IT 17% 47% 77% Natural 10% 7% 40% 天然的 *Gilts received 1-2cc dose of FluSure XP/RespiSure at 16 weeks of age 后备猪在 16 周时接种 1-2cc 的 FluSure XP/RespiSure **S:P cut-off of 1.2 for positive as result of natural exposure 自然接种的 S:P 值 1.2 被认为是阳性

21 But 1.5 yrs later, it was impossible to find suitable natural seeders and shortly thereafter we had clinical mature gilts with Mhp again. 但一年半以后, 我们未找到合适的天然种子, 不久之后, 我们又出现临床发病的后备母猪

22 2 nd Attempt: 第二次尝试 Use intra-tracheal inoculation on every group 每组均使用气管内接种 1 Intra-tracheal (IT) seeder : 21 Naïve pen mates 1 气管内免疫源播种 :21 个阴性栏

23 Confirm Mhp colonization 确定 Mhp 感染 Laryngeal swabs collected 6 weeks postexposure 接种 6 周后收集喉拭子 30 swabs, pools of 3 30 个拭子, 共 3 组 30-60% of pools Mhp PCR positive 各组 30-60% 的 Mhp PCR 检测为阳性 Ct s range 26.31, Ct 值范围为 26.31,36.33

24 Clinical signs following Mhp exposure vary by herd health status Mhp 接种后的临床症状因群体健康状况而异 Gilts from PRRS/Mhp positive herd 来自 PRRS/Mhp 阳性畜群的后备母猪 750 pigs/week 750 头猪 / 周 1-2% Thumping 1-2% 撞栏 1-3% Cough 1-3% 咳嗽 1-3% Fall-behinds 1-3% 发育迟缓 Signs variably responsive to antibiotics 对抗生素反应症状不同 Gilts from PRRS/Mhp negative herd 来自 PRRS/Mhp 阴性畜群的后备猪 650 pigs/week 650 头猪 / 周 No thumping 无撞栏 2% Cough occurs 3 weeks after seeders introduced 接种后 3 周 2% 出现咳嗽 No Fall-behinds 无迟缓 Cough responsive to antibiotics 对抗生素产生反应 ( 咳嗽 )

25 How will we determine success of Mhp acclimation process? 我们如何判定 Mhp 驯化过程是否成功? Improved reproductive performance of flows that historically received problematic gilts 改善历史上有问题的后备母猪流的繁殖表现 Pod 105, 101 breeding gilt performance improved during period entering Mhp exposed gilts 畜群 105,101 的后备母猪的育龄表现在 Mhp 暴露给后备猪的期间有所改善 Non-clinical commercial pig flow 非临床商业猪流动 Oral fluid monitoring of commercial pigs 对商品猪的口腔液体监测 Pod 601/602 (sourced by Pod 105)-1 positive case 群 601/602( 来源于群 105)- 阳性 1 例 Pod 201/202 (sourced by Pod 101)-negative 群 201/202( 来源于群 101)- 无阳性样本

26 References 参考文献 Anderson, et al. IPVS. 2016; 253. Desrosiers. JSHAP. 2001; 9: Lowe. Leman Conf. 2012; 83. Maes, et al. Vet Micro. 2008; 126: Pieters, et al. Vet Micro. 2009; 134: Pieters, et al. Can J Vet Res. 2010; 74: Pieters, et al. Leman Conf. 2013; 233. Pieters and Fano. Vet Record. 2016;178: Roos, et al. Vet Micro. 2016; 184: Ruiz, et al. JSHAP. 2003; 11: Villareal, et al. Vet J. 2011; 118:

鼠　疫（Plague）

鼠　疫（Plague）鼠疫 (Plague) 一疾病概述 (Disease description) 存在囓齒類及其跳蚤的一種人畜共通傳染病, 並藉跳蚤傳染給各種動物及人類, 其最初反應為跳蚤咬傷部位臨近的淋巴腺發炎, 這就是腺鼠疫, 經常發生於鼠蹊部, 少發生於腋下或頸部,