Matt Yarnall BVM&S MRCVS注册兽医师Cattle Veterinary AdviserUK 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

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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 Adviserfor the UK and Ireland英国和爱尔兰兽医顾问

Who am I? 我是谁？

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

Ubrolexin profile 优孢欣掠影o 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

o Treatment of clinical mastitis in lactating dairy cows for bacteria susceptible to the combination of cefalexin and kanamycin such as Staphylococcus aureus, Streptococccus agalactiae,Streptococcus dysgalactiae, Streptococcus uberis and Escherichia coli and CNS.

o 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o One tube. Two times. 24hrs apart. 仅需用药两次间隔 24 小时

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Why combine anything? 为什么组合

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

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The Antimicrobial Spectrum 抗菌谱 o Broad versus narrow o 广谱 VS 窄谱o Ubrolexin has a broad spectrum arising from 2 narrow

spectrum activeso 因为由两种窄谱抗生素相组合， Ubrolexin 抗菌谱广泛o Each active very focused on a specific group of pathogenso 每种抗生素特定作用于一部分病原菌

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

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What does synergy mean? 协同作用意味着什么o Synergy (from the Greek syn-ergo, meaning working

togethero 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

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Working together 协作

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Antibiotic synergy 抗生素的协同作用o 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)

o 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)

o In Ubrolexin®, the effect of Cefalexin is enhanced by Kanamycin and vice versa

o 对于 Ubrolexin 来说，卡那霉素可以加强头孢氨苄的作用，反之亦然

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o 1st generation cephalosporino第一代头孢类抗生素o Mainly active on Gram+o主要作用于革兰氏阳性菌o 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

o Aminoglycoside 氨基糖苷类抗生素o Active on Gram- and S. aureus o 主要作用于革兰氏阴性菌和金葡菌o Bactericidal activity through inhibition of bacterial

protein synthesis o 通过阻断细菌蛋白质的合成以达到杀菌目的

Kanamycin 卡那霉素

Cefalexin 头孢氨苄

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Mécanisme d´action

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How does it work? 作用机制o Cefalexin is active on the cell-wall of the bacteria altering its

permeabilityo 头孢氨苄作用于细菌细胞壁改变细胞壁的通透性o Cefalexin enhances the intracellular uptake of Kanamycin and

the access to its intracellular target, the 30s ribosomal subunito 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o Kanamycin decreases the protein synthesiso 卡那霉素减少蛋白质的合成o This results in a mutual enhancement of their respective

antimicrobial activityo 两种抗生素互相加强彼此的抗菌能力

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

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Prevent resistant strains 防止耐药菌株生成o 2 actives with different modes of action

• Cefalexin interferes with the cell wall• Kanamycin interferes with the genetic message2 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

o Less chance of resistance as bacteria must resist both modes of action

o 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

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Prudent use of antibiotics 严谨使用抗生素o Low level of resistance reported for 1st generation cephalosporins and for

Kanamycino 据报道，第一代头孢菌素和卡那霉素产生的耐药性都很低o Kanamycin is not widely used in cattleo 卡那霉素在牛上的应用不广泛o The development of resistance to both antibiotics would require at least two

mutations to occur simultaneously to prevent both antibiotics workingo 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o The enhanced killing activity of Cefalexin and Kanamycin has a beneficial

effect on slowing individual resistance developmento 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

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How can synergy be proven? 如何证明协同作用？o 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

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MIC (µg/ml)

Species 细菌种类 Cefalexin

头孢氨苄Kanamycin

卡那Combo 1.25:1

1Combo 1:2.3

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S. aureus 2 4 0,56 : 0,44 0,6 : 1,4S.aureus 2 4 1,11: 0,89 0,3 : 0,7Strep. dysgalactiae 0,5 128 0,56 : 0,44 0,6 : 1,4

Strep. dysgalactiae 0,5 64 0,56 : 0,44 0,6 : 1,4

Strep. uberis 0,5 16 0,14 : 0,11 0,15 : 0,35

Strep. uberis 0,25 32 0,28 : 0,22 0,3 : 0,7

E.coli 16 4 1,11 : 0,89 2,42 : 5,58

E.coli 16 4 2,22 : 1,78 0,6 : 1,41 around 3 hours after administration 用药后 3 小时2 around 6 hours after administration 用药后 6 小时

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随着时间的变化两种抗生素配比的变化

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How can synergy be proven? 如何证明协同作用o MIC = inhibitory data

• synergy can be measured in vitro by comparing the MICs of Cefalexin, Kanamycin and their combination

o 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

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Kill kinetics 杀菌动力曲线

Effect of antimicrobial combinations抗生素组合的效果

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Assessing antibiotic synergy 抗生素协同作用评估o Synergy can be measured in vitro by comparing the kill

kinetics of Cefalexin, Kanamycin and their combinationo 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o Time-kill curves offer dynamic information about the

bactericidal activity of an antibiotic combination over timeo 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

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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

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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

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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

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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）

At every level of the udder 在乳腺组织中的各层分布

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o Tissue distribution.mov

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Post Antibiotic Effect (PAE) 抗生素后效应o Persistent suppression of bacterial growth even at

concentrations below the MICo 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o Drugs that demonstrate PAE require less frequent

administrations than those that do noto 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o 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

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1

10

100

1000

10000

0 1 2 3 4 5 6Time (hours)

Viab

le C

ount

(cfu

/ml)

Control

1.6 hours to increase 1 log10

+ 1 log10

3.1 hours to increase 1 log10

Antibiotic induced death

Example 示例

Removal of antibiotic

In this example, PAE = 3.1 - 1.6 = 1.5 hours 在此例， PAE= 3.1 - 1.6 = 1.5 小时

+ 1 log10

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PAE on Staphylococcus aureus 针对金葡菌的抗生素后效应

Drug 药物 Concentration (µg/ml)

浓度PAE (h)

后效应Cephalexin 头孢氨苄

8 2,4

Kanamycin卡那霉素

4 4,8

Ubrolexin®优孢欣 ®

1,2 : 2,8 6,1

Post Antibiotic Effect抗生素后效应

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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

Post Antibiotic Effect

Species Concentration(µg/ml)

PAE (h)

PASME (h)

(¼ MIC)

PASME (h)

(½ MIC)Strep. Uberis乳房链球菌

0.6 : 1.4 1.6 3.7 4.3

E. Coli 大肠杆菌 2.4 : 5.6 1.1 1.6 6.5S. Aureus 金葡菌 1.2 : 2.8 6.1 5.7 8.5Strep. Dysgalactiae停乳链球菌

2.4 : 5.6 4.2 6.2 > 10.2

Strep. Agalactiae无乳链球菌

2.4 : 5.6 3 3.8 6.9

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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o Ubrolexin® exerts larger PAE and PASME against all target mastitis

pathogens compared with the individual compoundso 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

o This supports the once-a-day treatment regimeo 再一次支持了一天一次用药的治疗方案

Post Antibiotic Effect

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Conclusion 结论o Ubrolexin® exhibits synergistic activity against key mastitis

pathogens, in MHB, in milk and in udder tissueo 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o 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

o 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o Through its synergistic activity, Ubrolexin® kills major pathogens

within 9-12 hourso 通过这种协同作用， Ubrolexin 可以在 9 － 12 小时内杀死主要致病菌

Kill kinetics杀菌动力曲线

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Field efficacy data 田间数据o 2 multi-centre field

studieso 2 个研究中心的田间研究

• Ubrolexin® vs. cefquinome in the UK and France

• 优孢欣 ® 对比头孢喹肟• 英国和法国• Ubrolexin® vs. cefoperazone

in the UK and Germany• 优孢欣 ® 对比头孢哌酮• 英国和德国

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

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

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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

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Ubrolexin® - the routine treatment of clinical mastitis

优孢欣 ® - 临床乳房炎的常规治疗用药

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Cefalexin and Kanamycin 头孢氨苄和卡那霉素o 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

o 2 complementary antibacterial activity 两种互补的抗菌活性o Working in synergy 协同工作o Delivering enhanced and faster bactericidal activityo 完成更强更快的杀菌效果

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Ubrolexin® - the power of synergy 优孢欣 ® - 协同的力量o 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

o Approved to the latest European standardso 符合欧洲最新标准o Suitable for routine use in mastitis treatmento 适合于乳房炎的常规治疗

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Ubrolexin® - clever and convenient 优孢欣 ®- 高效便捷o Ubrolexin® offers fast killing activity and prolonged PAEo 优孢欣 ® 杀菌更快，抗生素后效应更长o Ubrolexin® therefore requires less administrations (one per

day) to deliver the right dose for maximum efficacyo 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

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Questions?