Impact of less antimicrobial usage on diagnostics of infectious

Back to the future – Diagnostic considerations in a growth promoters-free industry

回到未来 —不用生长促进剂的养殖业的兽医诊断

Fabio Vannucci （法比奥.万努奇）

Outline

• History历史

• EU experiences欧盟经验

o Disease incidence发病率

• USA scenario美国情况

o Diagnostics诊断

• USA and EU: compare/contrast 美国和欧盟：对比/比较

• Considerations for future scenarios将来的注意事项

Overview 概要

• Use of antimicrobial growth promoter (AGP) 抗菌生长促进剂(AGP)的使用

- Late 1940s: Chickens fed with fermentation waste from tetracycline production as a source of vit B12 20世纪40年代后期：鸡饲喂生产四环素发酵废弃物作为维生素B12的来源

Overview History历史

- Better growth performance due to residual tetracycline: “Animal protein factor”

由于残留四环素”动物蛋白因子”，产生更好的生长成绩。

- Early 1950 (Cunha et al, Luecke et al): 20世纪50年代早期

o The effect of vitamin B12, animal protein factor and streptomycin on the growth of young pigs

维生素B12、动物蛋白因子和链霉素对仔猪的生长产生影响。

• Use of antimicrobial growth promoter and antimicrobial resistance 使用抗菌促生产剂和耐药性


- Inaction (UK and other EU countries): tylosin, spiramycin, avoparcin… 无行动(英国和其他欧盟国家):泰乐菌素、螺旋霉素、阿伏霉素...

- Action (Sweden): consumers → Federation of Swedish Farmers → ban in 1986 行动(瑞典):消费者→瑞典农民联合会→1986年禁止

- Mid 1960 (UK): Multi-drug-resistant salmonella 60年代(英国):多重耐药性沙门氏菌

- “Sufficient sound basis for action” 采取行动的充分可靠依据”

- Recommendations for using on animal feed:饲料中使用抗生素的建议 o Little or none application for therapeutic purposes in human or animals在人和动物上少用或不用治疗用抗生素。

o Not impair the efficacy of a prescribed therapeutic drugs through development of resistance 通过耐药性的形成,但不破坏处方治疗药物的疗效

o Tylosin should be available only for therapeutic purposes 泰乐菌素应该只提供给治疗用。

• Use of antimicrobial growth promoter and antimicrobial resistance 使用抗菌促生长剂和耐药性


- Vancomycin (glycopeptide antimicrobial): discovered early 1960s 万古霉素(糖肽类抗生素):发现20世纪60年代初 o 1980s: widely used in humans for treatment of resistant Gram positive bacteria

80年代广泛应用用于治疗人的耐药革兰氏阳性菌。 o 1993: Vancomycin resistant enterococci isolated from humans and animals (Bates et al 1993)

1993年从人和动物分离出耐万古霉素肠球菌 Not been used in food animal食品动物中没有使用 o Cross-resistance with Avoparcin: commonly used for growth promotion in food animal与阿伏霉素有交叉耐药:常用于食品动物促生长

EU banned in 1997 欧盟1997明禁用 - AGP termination 动物生长促进剂(AGP)停用时间

Sweden瑞典: 1986 Norway挪威: 1995 Switzerland瑞士: 1999 Finland芬兰: 1999 Denmark丹麦: 2000 EU欧盟: 2006

Overview European experiences: disease incidence 欧洲经验:发病率

• How to measure disease incidence (before/after AGP ban)? 如何测量发病率（AGP禁用前后）

- Indirect method for occurrence of disease 疾病发生的间接方法 o Antimicrobial usage ONLY for therapeutic or preventive purposes (assumption after AGP ban) 假设禁用AGP后,抗生素仅用在治疗或预防上

o Recording antimicrobial usage for a given clinical syndrome记录针对指定临床综合征使用的抗生素

- Production data: mortality, ADG, Feed conversion生产数据:死亡率\平均日增重\饲料转化

- Diagnostic data (lab accessions)诊断数据(实验室病例) o High specificity but questionable sensitivity 高特异性,但敏感性有问题 o Confounders - emerging disease, surveillance, detection vs disease 混杂因素 - 新出现疾病，监测，检测 VS 疾病



Denmark丹麦

- Production data: mortality, ADG, Feed conversion 生产数据:平均日增重,平均日增重,饲料转化率

平均日增重

死亡率

断奶猪禁用促生长剂

时间(年)



- Overall trends on antimicrobial consumption (therapeutic use) 抗生素使用(治疗用)的总体趋势

Sweden瑞典 Norway挪威


• How to measure disease incidence (before/after AGP ban)? 如何评估发病率（AGP禁用前后）

- Overall trends on antimicrobial consumption (therapeutic use) 抗生素使用(治疗用)的总体趋势

Denmark丹麦 Netherlands荷兰



- Age groups 年龄组

Sweden (1986) – first 4 years after 瑞典(1986年)—禁用AGP后的头4年

o Increase post-weaning diarrhea (> 1.5% mortality) 增加断奶后腹泻(>1.5%死亡率)

o 5-6 more days to reach 25 kg 达25Kg体重增加5-6天

o ↓ 1.5% feed efficiency in finishers育肥阶段饲料转化率↓ 1.5%

母猪和仔猪断奶仔猪

育肥猪总体,校正后



- Antimicrobial classes (Denmark) 抗生素分类(丹麦)



- Antimicrobial classes (Netherlands) 抗生素分类(荷兰)

Switzerland

瑞士



- Clinical signs, age group, # days and animals treated 临床症状\年龄组,动物治疗和天数

腹泻

月: 相对于不用AGP

肺炎

关节炎消瘦

其他疾病

治疗比例

• Enteric pathogens after AGP ban 上禁用AGP后的肠道病原

o Significant increase in diarrhea (nursery) 保育腹泻明显增加

- Post-weaning diarrhea: β-hemolytic E. coli 断奶后腹泻: β溶血大肠杆菌

- Spirochetal colitis (Brachyspira pilosicoli) 螺旋体结肠炎(多毛结肠短螺旋体)

- Clinical ileitis (Lawsonia intracellularis) 临床回肠炎（胞内罗松菌）

• ↓50-120g ADG 平均日增重↓50-120g • ~ 7 additional days from weaning to 30Kg断奶至30K多7天

Overview European experience: disease incidence 欧洲经验:发病率

Weber et al (2016), 24th IPVS

1 days 5 days 2 weeks 3 weeks 5 weeks 10 weeks 28 weeks Adult

Escherichia coli大肠杆菌

Brachyspiral colitis螺旋体结肠炎

Lawsonia intracellularis胞内罗松菌

Neonatal亲生 Nursery保育 Finishing育肥

Overview

Autolysis自溶 Non-autolysis非自溶

USA Scenario: Diagnostics美国情况:诊断

• Enterotoxigenic Escherichia coli (ETEC)肠毒大肠杆菌

o Post-weaning diarrhea: β-hemolytic E. coli 断奶后腹泻: β溶血大肠杆菌

o High morbidity and variable mortality高发病率和死亡率差异大

Overview

o Virulence毒力 - Fimbria (F18, F4) + Enterotoxins (LT, STa, STb, EAST1, Stx2e, AIDA)

纤毛（F18,F4)+ 肠毒素(LT, STa, STb, EAST1, Stx2e)

- Stx2e: Sudden death (CNS signs may be present) 突然死亡(中枢神经症状可能会出现)

USA Scenario: Diagnostics美国情况:诊断 β-溶血

α-溶血 γ-溶血

• Enterotoxigenic Escherichia coli (ETEC) 肠毒大肠杆菌

Overview USA Scenario: Diagnostics美国情况:诊断

UMN-VDL: Enterotoxigenic E. coli 2010-2016 (n=2206) 明大-兽医诊断实验室-肠毒大肠杆菌 2010-2016(2206例)

Overview

F18 F4

Neg

Others (F41, F5, F6)

F18.LT

F18.STb

F18.STX2e

F4.LT

F4.STa F4.STb

F4. STa.STb

F4.LT.STa.STb

Weber et al (2016), 24th IPVS


UMN-VDL: Antimicrobial susceptibility (2006-2016)抗生素敏感性

Overview β-

hem

olyt

ic E

. col

i β-溶

血大

肠杆

菌

Ampicillin氨苄青霉素

Enrofloxacin恩诺沙星 Trimethoprim/SulphaTMP/磺胺 Gentamicin庆大霉素 Neomycin新霉素 Florfenicol氟苯尼考

Sulpha磺胺

Oxytetracycline土霉素

Tiamulin泰妙灵 Chlortetracycline金霉素


• Brachyspiral colitis短螺旋属结肠炎

Overview

“B. suanatina” Swine Swine dysentery-like “B. hampsonii” Swine Swine dysentery-like

Strongly

β-hemolytic

Weakly

β-hemolytic


弱β-溶血

强β-溶血

种类主要宿主主要疾病

UMN-VDL: B. pilosicoli isolation (2011-2016)多毛结肠短螺旋体的分离

Overview

0

1

2

3

4

5

6

7

8

9

Num

ber o

f B. p

ilosi

coli


UMN-VDL: Antimicrobial susceptibility (2009-2013)抗生素敏感性


卡巴氧沃尼妙林泰妙灵强力霉素林可霉素泰乐菌素

Sweden: Antimicrobial susceptibility (AGP free scenario) 瑞典:抗生素敏感性(无AGP情况)


• Proliferative enteropathy (ileitis): Lawsonia intracellularis 增生性肠炎(回肠炎):胞内罗松菌

Overview

o Endemically distributed worldwide流行分布于全世界

o Classically affected growing-finishing pigs 常见于生长育肥猪爱影响

0

10

20

30

40

50

60

70

2 5 7 10 13 16 19 22Weeks周

0

10

20

30

40

50

60

70

5 7 10 13 16 19 22Weeks周

Feca

l PC

R粪

便PC

R

Sero

logy

血清

学

o Clinical presentation (Acute/Chronic/Subclinical) 临床表明(急性/慢性/亚临床）


• Proliferative enteropathy (ileitis): Lawsonia intracellularis 增生性肠炎(回肠炎):胞内罗松菌 o Clinical/subclinical PCR threshold to justify the treatment (?) 临床/亚临床 PCR检测来证实处理(?)

Overview

1.00E+00

1.00E+01

1.00E+02

1.00E+03

1.00E+04

1.00E+05

1.00E+06

1.00E+07

1.00E+08

1.00E+09

1.00E+10

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42

Log 1

0 Law

soni

a in

trac

ellu

laris

bac

teria

/g fe

ces

Ct value Ct值

UMN-VDL

- One Log10 increase in LI load increases OR for a pig to have a low growth rate by 2 times


Overview

UMN-VDL: Lawsonia intracellularis PCR detection - 2013-2016 (n=3577)明大-兽医诊断实验室:胞内罗松菌的PCR检测-2013-2016(3577例)

10

12

14

16

18

20

22

24

26

28

30

32

34

36

38

40

42

10/3/2013 4/21/2014 11/7/2014 5/26/2015 12/12/2015 6/29/2016

Ct v

alue

25%

34%

41%

USA Scenario: Diagnostics美国情形:诊断

Overview

UMN-VDL: Antimicrobial susceptibility抗生素敏感性

USA Scenario: Diagnostics美国情形:诊断

• Increasing of enteric bacterial infection (not much respiratory)肠道细菌感染增加(呼吸道菌增加不多)

Overview USA and EU: compare/contrast 美国和欧洲:比较/对比

o Porcine Respiratory Disease Complex (PRDC)猪呼吸首综合征 - Success controlling primary triggers – vaccination, elimination 成功控制的主要手段--免疫,净化

o Microbiome complexity 微生物的复杂性

o Porcine Enteric Disease Complex (PEDC) – What the heck?猪肠道疾

病综合征--到底怎么回事 - Enteric bacteria as primary cause肠道细菌是主要的原因 - Synergism: Rotavirus – Clostridium perfringens, E.coli (?) 协同:轮状病毒 - 产气荚膜梭菌,大肠杆菌(?)

Overview

Europe欧洲 United States美国 Weaning age断奶日龄 28-30 days 21 days PRRS status蓝耳状况 Neg (some countries)阴 Pos阳 Pig/farm density猪/场密度 Low/variable低/差异大 High高 Lawsonia vaccine usage罗松菌疫苗使用情况 Low低 Widely used使用广 Carbadox卡巴氧 Banned禁止 Widely used*使用广

Zinc oxide氧化锌 Partial restrictions部分地方限用

Widely used使用广

USA and EU: compare/contrast 美国和欧洲:比较/对比

• Management/alternative strategies管理/备选策略

• Systematic approach to measure disease incidence系统方法评估发病率

• Scale matters 规模方面

• AGP ban (EU)禁用AGP(欧盟) – Enteric bacterial pathogens (diarrhea): When to act (?) 肠道细菌病原

(腹泻):什么时候开始采取措施(?) • Lawsonia PCR positive: “how much” to justify treatment (?) and for how long (?)罗松菌PCR阳性:”多少”需要确认治疗(?)和治疗多久(?)

• Beta-hemolytic E coli with virulence factor + diarrhea β-溶血大肠杆菌带有毒力因子引起腹泻

Outline Considerations for future scenarios 将来的注意要点

– Followed by additional regulation → Long term benefits 接着是更多的监管→长期受益

• Denmark (Yellow card regulations)丹麦（黄牌规章制度） • Germany (DART, German Antibiotic Resistance Strategy)德国（DART，德国耐药性策略）

• Netherlands (Netherlands Veterinary Medicines Authority - SDa)荷兰（荷兰兽药管理局-SDa)

Outline

• Talking about additional regulation…谈论更多的监管...

Considerations on future scenarios 将来的注意要点

©2014 Regents of the University of Minnesota. All rights reserved. The University of Minnesota is an equal opportunity educator and employer.

• Dr. Peter Davies

• Dr. Connie Gebhart

• Dr. Julio Alvarez

Aknowledgements Acknowledgements 致谢

Fabio Vannucci [email protected]

Documents

Impact of less antimicrobial usage on diagnostics of infectious