21 st Century Mastitis Treatment 21 世纪乳房炎治疗

1. Matt Yarnall BVM&S MRCVS 注册兽医师 Cattle Veterinary Adviser UK and Ireland 英国和爱尔兰牛兽医顾问 21st Century Mastitis Treatment 21世纪乳房炎治疗 China May 2013

2. Who am I? 我是谁？ • Matt Yarnall BVM&S MRCVS 执业兽医师 • Worked in practice as a mixed vet • and dairy vet 曾任职多畜种兽医 • Cattle Veterinary Adviser • for the UKand Ireland • 英国和爱尔兰兽医顾问

3. What am I here to talk to you about?在这里和大家一起分享？ • What is Ubrolexin®? • 优孢欣是什么？ • Treatment with Ubrolexin® – How does it work? • 用优孢欣进行治疗－它如何起到效果？ • The success of Ubrolexin® – Some field studies • 优孢欣的成功案例－田间试验研究

4. Ubrolexin profile优孢欣掠影 • Broad spectrum intramammary tube广谱乳区注入剂 • Cefalexin头孢氨苄 ................................... 200 mg • Kanamycin卡那霉素 ................................ 100 MUI • Treatment of clinical mastitis in lactating dairy cows for bacteria susceptible to the combination of cefalexin and kanamycin such as Staphylococcus aureus, Streptococccusagalactiae,Streptococcusdysgalactiae, Streptococcus uberisand Escherichia coli and CNS. • 头孢氨苄与卡那霉素的组合用于治疗对金葡菌，无乳链球菌，停乳链球菌，乳房链球菌，大肠埃希氏菌属及凝固酶阴性葡萄球菌敏感的临床性乳房炎 • One tube. Two times. 24hrs apart. 仅需用药两次间隔24小时

5. Why combine anything?为什么组合

6. Rationales for antibiotic combinations抗生素组合的原理 • Broaden the antimicrobial spectrum • 扩大抗菌谱 • Take advantage of potential synergism • 利用潜在的协同作用 • Prevent emergence of resistant strains • 预防耐药菌株的出现

7. The Antimicrobial Spectrum 抗菌谱 • Broad versus narrow • 广谱 VS 窄谱 • Ubrolexin has a broad spectrum arising from 2 narrow spectrum actives • 因为由两种窄谱抗生素相组合，Ubrolexin抗菌谱广泛 • Each active very focused on a specific group of pathogens • 每种抗生素特定作用于一部分病原菌

8. Why Combine? 为什么要组合（联合用药） • Broaden the antimicrobial spectrum • 扩大抗菌谱 • Take advantage of potential synergism • 利用潜在协同作用 • Prevent emergence of resistant strains • 防止耐药菌株的出现

9. What does synergy mean? 协同作用意味着什么 • Synergy (from the Greeksyn-ergo, meaning working together • 协同（来自于希腊语，意味着一起工作） • is the term used to describe a situation where the final outcome of a system is greater than the sum of its parts • 用来表述系统共同作用的最终结果大于每个部分独立作用的加和

10. Working together 协作

11. Antibiotic synergy 抗生素的协同作用 • The enhanced result of positive interactions between two antimicrobial agents so that their combined effect is greater than the sum of their individual effects (1 + 1 = 3) • 两种抗菌成分之间产生积极的作用从而使协同作用的结果大于各个部分独立作用的总和 (1 + 1 = 3) • In Ubrolexin®, the effect of Cefalexin is enhanced by Kanamycin and vice versa • 对于Ubrolexin来说，卡那霉素可以加强头孢氨苄的作用，反之亦然

12. 1st generation cephalosporin • 第一代头孢类抗生素 • Mainly active on Gram+ • 主要作用于革兰氏阳性菌 • Bactericidal activity by inhibition of bacterial cell wall synthesis 通过抑制细菌细胞壁的合成来达到杀菌效果 Cefalexin头孢氨苄 Kanamycin卡那霉素 • Aminoglycoside氨基糖苷类抗生素 • Active on Gram- and S. aureus • 主要作用于革兰氏阴性菌和金葡菌 • Bactericidal activity through inhibition of bacterial protein synthesis • 通过阻断细菌蛋白质的合成以达到杀菌目的 12

13. Mécanisme d´action

14. How does it work? 作用机制 • Cefalexin is active on the cell-wall of the bacteria altering its permeability • 头孢氨苄作用于细菌细胞壁改变细胞壁的通透性 • Cefalexin enhances the intracellular uptake of Kanamycin and the access to its intracellular target, the 30s ribosomal subunit • 头孢氨苄协助卡那霉素进入细胞并作用于靶器官核糖体30s亚基 • Kanamycindecreases the protein synthesis • 卡那霉素减少蛋白质的合成 • This results in a mutual enhancement of their respective antimicrobial activity • 两种抗生素互相加强彼此的抗菌能力

15. Why Combine? 为什么要联合用药？ • Broaden the antimicrobial spectrum • 扩大抗菌谱 • Take advantage of potential synergism • 利用潜在的协同作用 • Prevent emergence of resistant strains • 防止耐药菌株的出现

16. Prevent resistant strains 防止耐药菌株生成 • 2 actives with different modes of action • Cefalexin interferes with the cell wall • Kanamycin interferes with the genetic message 2种活性成分有着不同的作用机制 头孢氨苄干扰细胞壁合成 卡那霉素干扰遗传信息表达 • Less chance of resistance as bacteria must resist both modes of action • 由于细菌很难同时抵抗两种抗生素作用模式，减少了抗药性的产生

17. Prudent use of antibiotics 严谨使用抗生素 • Low level of resistance reported for 1st generation cephalosporinsand for Kanamycin • 据报道，第一代头孢菌素和卡那霉素产生的耐药性都很低 • Kanamycin is not widely used in cattle • 卡那霉素在牛上的应用不广泛 • The development of resistance to both antibiotics would require at least two mutations to occur simultaneously to prevent both antibiotics working • 产生对这两种抗生素同时耐药，需要至少两种突变模式形成 • The enhanced killing activity of Cefalexin and Kanamycin has a beneficial effect on slowing individual resistance development • 头孢氨苄和卡那霉素共同作用形成的杀菌效果，抑制了耐药性的形成

18. How can synergy be proven? 如何证明协同作用？ • MIC (minimum inhibitory concentration) = inhibitory data 最小抑菌浓度=抑菌数据 • synergy can be measured in vitro by comparing the MICs of Cefalexin, Kanamycin and their combination • 通过对头孢氨苄、卡那霉素与其组合进行最小抑菌浓度比较可以在体外衡量协同作用

19. MIC (µg/ml) 1 around 3 hours after administration 用药后3小时 2 around 6 hours after administration 用药后6小时

20. 随着时间的变化两种抗生素配比的变化

21. How can synergy be proven? 如何证明协同作用 • MIC = inhibitory data • synergy can be measured in vitro by comparing the MICs of Cefalexin, Kanamycin and their combination • The killing curve = dynamic picture • bactericidal activity of an antibiotic combination over time = no. of survivors after administration of drug (s) 杀菌曲线＝动态图示 随着时间变化抗生素组合的杀菌作用 ＝用药后残存的细菌数

22. Kill kinetics 杀菌动力曲线 Effect of antimicrobial combinations 抗生素组合的效果

23. Assessing antibiotic synergy 抗生素协同作用评估 • Synergy can be measured in vitro by comparing the kill kinetics of Cefalexin, Kanamycin and their combination • 可通过体外比较头孢氨苄，卡那霉素，抗生素组合的杀菌动力曲线来证明协同作用 • Time-kill curves offer dynamic information about the bactericidal activity of an antibiotic combination over time • 时间－杀菌曲线可以动态体现抗生素组合随着时间变化的杀菌活动

24. Ganiere KK study 杀菌动力研究 通过时间－杀菌曲线比较头孢氨苄、卡那霉素及合成物对抗金黄色葡萄球菌的最低抑菌浓度，从而看到协同杀菌的效果

25. Ganiere KK study 杀菌动力研究 通过时间－杀菌曲线比较头孢氨苄、卡那霉素及其合成物对抗乳房链球菌最低抑菌浓度，从而看到协同杀菌的效果

26. Ganiere KK study 杀菌动力研究 通过时间－杀菌曲线比较头孢氨苄、卡那霉素及其合成物对抗大肠杆菌最低抑菌浓度，从而看到协同杀菌的效果

27. Ganiere KK study – in milk 杀菌动力研究－牛奶中 不同浓度头孢氨苄：卡那霉素对抗金葡菌的时间－杀菌曲线（牛奶中）

28. Ateverylevel of the udder在乳腺组织中的各层分布 • Tissue distribution.mov

29. Post Antibiotic Effect (PAE) 抗生素后效应 • Persistent suppression of bacterial growth even at concentrations below the MIC • 即使抗生素浓度低于最低抑菌浓度MIC，仍然可以持续的抑制细菌生长 • Drugs that demonstrate PAE require less frequent administrations than those that do not • 有抗生素后效应（PAE）的药物同没有PAE的抗生素相比，给药频率更低 • This supports the once-a-day treatment regime这一点有效支持了一天一次的用药程序

30. 1.6 hours to increase 1 log10 + 1 log10 + 1 log10 3.1 hours to increase 1 log10 Example示例 Antibiotic induced death Removal of antibiotic Viable Count (cfu/ml) Control In this example, PAE = 3.1 - 1.6 = 1.5 hours 在此例，PAE= 3.1 - 1.6 = 1.5 小时

31. Post Antibiotic Effect 抗生素后效应 PAE on Staphylococcus aureus针对金葡菌的抗生素后效应

32. Post Antibiotic Effect PAE and PASME of Ubrolexin® 优孢欣的抗生素后效应及亚最小抑菌浓度后效应

33. Post Antibiotic Effect PAE and PASME of Ubrolexin® 优孢欣的抗生素后效应及亚最小抑菌浓度后效应 • Ubrolexin®exerts larger PAE and PASME against all target mastitis pathogens compared with the individual compounds • 与单独应用头孢氨苄和卡那霉素相比，优 孢欣对主要乳房炎致病菌展示了明显的抗生素后效应及亚最小抑菌浓度后效应 • PAE range 1.1 – 6.1 hrs • PASME range 1.6 - >10 hrs • This supports the once-a-day treatment regime • 再一次支持了一天一次用药的治疗方案

34. Kill kinetics 杀菌动力曲线 Conclusion 结论 • Ubrolexin®exhibits synergistic activity against key mastitis pathogens, in MHB, in milk and in udder tissue • Ubrolexin对主要的乳房炎致病菌（培养基、牛奶中和乳腺组织内）显示了协同作用 • This synergy results in a larger and faster rate of kill compared to the single antibiotics alone, while a lower total amount of antibiotic is used • 这种组合与各抗生素单独作用相比杀菌效果更快，杀菌谱更广，并且抗生素用量小 • Through its synergistic activity, Ubrolexin®kills major pathogens within 9-12 hours • 通过这种协同作用，Ubrolexin可以在9－12小时内杀死主要致病菌

35. Field efficacydata 田间数据 • 2 multi-centre fieldstudies • 2个研究中心的田间研究 • Ubrolexin® vs. cefquinomein the UK and France • 优孢欣®对比头孢喹肟 • 英国和法国 • Ubrolexin® vs. cefoperazonein the UK and Germany • 优孢欣®对比头孢哌酮 • 英国和德国

36. Isolates were representative of usual field infections 现场分离的细菌类型

37. Bacteriological cure rates achieved with Ubrolexin®match expectations 优孢欣取得的细菌学治愈率符合预期

38. Quarters treated with Ubrolexin®or cefquinome were not significantly different from each other, but were significantly more likely to be pathogen free post treatment than cefoperazone-treated quarters (p=0.021) 治疗效果，优孢欣和头孢喹肟（第四代头孢菌素）相比，无明显差异。但明显好于第三代头孢菌素头孢哌酮

39. Ubrolexin® - the routine treatment of clinical mastitis 优孢欣® -临床乳房炎的常规治疗用药

40. Cefalexin and Kanamycin头孢氨苄和卡那霉素 • 2 complementary compounds 2种互补成分 • Different classes 不同种类 • Different modes of action 不同的作用方式 • Different pathogen targets 不同的靶致病菌 • 2 complementary antibacterial activity两种互补的抗菌活性 • Working in synergy 协同工作 • Delivering enhanced and faster bactericidal activity • 完成更强更快的杀菌效果

41. Ubrolexin®- the power of synergy 优孢欣® - 协同的力量 • Combination of 2 target-spectrum antibiotics to offer 两种不同抗菌谱抗生素的组合 • uncompromised and enhanced broad spectrum activity • 更广的抗菌谱 • Fast kill for highest efficacy and protection • 快速杀菌达到最佳效果，并提供最佳保护力 • Approved to the latest European standards • 符合欧洲最新标准 • Suitable for routine use in mastitis treatment • 适合于乳房炎的常规治疗

42. Ubrolexin®- clever and convenient 优孢欣®-高效便捷 • Ubrolexin®offers fast killing activity and prolonged PAE • 优孢欣®杀菌更快，抗生素后效应更长 • Ubrolexin®therefore requires less administrations (one per day) to deliver the right dose for maximum efficacy • 所以优孢欣®需要更少的剂量和注药次数即可达到最大的效果 • Less labour 更少的人工 • Less teat manipulation 乳头操作更少 • Lower contamination risk 感染风险更低

44. Questions?