1. Antimicrobial resistance (AMR) profiling in Escherichia coli from farmed and wild animals in the Mekong delta of Vietnam Nguyen Thi Nhung Oxford University Clinical Research Unit, HCMC, Vietnam Biodiversity and Health Symposium Phnom Penh, 17-18 Nov 2014

2. Agenda Introduction − Animal production in Mekong Delta of Vietnam Methods − Sampling − Laboratory methods Results − Prevalence of AMR E. coli from farmed and wild animals − AMR patterns of E. coli from different species − Risk factors for multidrug resistance Conclusion 2

3. Animal production in Mekong Delta of Vietnam Large poultry and pig population Mixed farms Small scale production systems including household backyard farms Duck production systems integrated with rice production systems (free ranging ducks) Low bio-security and bio-containment 3

4. Commensal bacteria of warm-blooded animals High prevalence, simple and efficient isolation procedures Useful indicator for the presence of AMR in monitoring and surveillance programs Use of Escherichia coli as indicator No studies have investigated the prevalence of AMR E. coli isolated from the farmed and wildlife species in the Mekong Delta of Vietnam 4

5. Aims of study To investigate the prevalence of AMR on E. coli indicator bacteria in various food animals and wildlife To characterize E. coli phenotypic resistant profiles To identify risk factors for fecal carriage of AMR E. coli in animal production 5

6. Cao Lanh district, Dong Thap province 90 pig, duck and chicken farms (30 of each) were stratified by size (small, medium and large) Fecal material was collected using a pair of boot swabs Farm survey design 6

7. Small wild mammals were trapped from pig and poultry farms, rice fields, forests Humanely culled by overdose of isoflurane 0.2g caecal contents were collected from large intestines Small wild mammals trapping and processing 7

8. Laboratory methods Antimicrobial agentsAcronym Amoxicilin/clavulanic acid (30µg)AMC Ampicillin (10µg)AMP Ceftazidime (30µg)CAZ Chloramphenicol (30µg)C Ciprofloxacin (5µg)CIP Gentamicin (10µg)CN Tetracycline (30µg)TE Trimethoprim-sulphamethoxazol (25µg)SXT AMR E. coli testing E. coli isolation 8

9. Results 9

10. E. coli isolates Species No. samples No. isolates Total isolates Farmed animals Chicken30148 434 Duck30143 Pig30143 Small wild mammals Rattus argentiventer933 234 Rattus tanezumi1962 Rattus exulans516 Rattus norvegicus1028 Bandicota indica1667 Suncus murinus728 Total 156 668 B. indica R. argentiventer R. exulans R. norvegicus R. tanezumi S. murinus 10

11. Prevalence of AMR E. coli from farmed animals TE: tetracycline, SXT: trimethoprim-sulphamethoxazole, CN: gentamicin, CIP: ciprofloxacin, AMP: ampicillin, AMC: amoxicilin- clavulanic acid, CAZ: ceftazidime, C: chloramphenicol, ESBL: extended spectrum β lactamase, MDR: multidrug resistance, was defined as resistant to at least three different of antimicrobials Unadjusted dataAdjusted data 11

12. Prevalence of AMR E. coli from small wild animals Farms Non-farms TE: tetracycline, SXT: trimethoprim-sulphamethoxazole, CN: gentamicin, CIP: ciprofloxacin AMP: ampicillin, AMC: amoxicilin- clavulanic acid, CAZ: ceftazidime, C: chloramphenicol, ESBL: extended spectrum β lactamase, MDR: multidrug resistance, was defined as resistant to at least three different of antimicrobials 12

13. Minimum spanning tree of AMR patterns from different species 13

14. DAPC of AMR profiles by host type DAPC= Discriminant Analysis of Principal Components 14

15. Multivariable logistic regression of MDR for isolates from farms OR95% CIp-value Type of farm (baseline=chicken or duck)1.0-- Small medium pig farm1.181.04-1.350.0104 Large pig farm1.411.27-1.57<0.001 Age farmer (per 10-year period)0.860.82-0.90<0.001 Well-water: Large pig farm1.241.15-1.33<0.001 Small medium pig farm1.551.37-1.75<0.001 15

16. Conclusions High levels of resistance to common antimicrobials among farmed animals − Extensive range of antimicrobial compounds used not according to the dosage, length of treatment of the manufacturers − Antimicrobials used on farm and human medicine are largely similar Small wild mammals trapped on farms seem to have higher prevalence of AMR − Wildlife contact directly with antimicrobial residuals or resistant bacteria through food and effluent systems − Useful indicators of the presence of antimicrobial resistant populations in the environment and/or farms There are some factors such as farm size (pigs), age of farmers, and water sources that may contribute to the problem of AMR on farms − The longitudinal studies need to be conducted 16

17. Acknowledgements: Dr. Juan Carrique-Mas Dr. Ngo Thi Hoa Mr. James Campbell Mr. Nguyen Van Cuong Ms. Vo Nhu Thanh Truc Dr. Serge Morand Sub-Department of Animal Health Dong Thap Hospital Tropical Diseases HCMC Funding: VIZIONS WT/093724 ZoNMW / WOTRO/205100012 (The Netherlands) Li-Ka-Shing Foundation 17

18. Thank you!