Cambodia AGISAR/WHO and FAO project,2012 Dr Kruy Sun Lay, Head of - - PowerPoint PPT Presentation

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Cambodia AGISAR/WHO and FAO project,2012 Dr Kruy Sun Lay, Head of - - PowerPoint PPT Presentation

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Implementation of AMR research in Cambodia AGISAR/WHO and FAO project,2012 Dr Kruy Sun Lay, Head of Food microbiology Lab Plan Project titles Overall project aims Chronology of events Poultry production in Cambodia Project

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Implementation of AMR research in Cambodia AGISAR/WHO and FAO project,2012

Dr Kruy Sun Lay, Head of Food microbiology Lab

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Plan

 Project titles  Overall project aims  Chronology of events  Poultry production in Cambodia  Project approach FAO/WHO

 Sample site and design  Poultry sample collection  Poultry slauther chain continuum  Sample size for FAO/WHO

 Project outputs  Preliminary results

Thailand Workshop 17-19/09/2012

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AGISAR/WHO and FAO project

I-WHO/“An Integrated surveillance study of antimicrobial resistance in Salmonella subspecies, Campylobacter spp, Escherichia coli and Enterococcus spp from poultry carcasses and Salmonella subspecies,Campylobacter spp from inpatients of National Pediatric Hospital, Phnom Penh, Cambodia” II-FAO/“the control of microbial and antimicrobial resistance hazards along the poultry chain continuum, in Cambodia”

Thailand Workshop 17-19/09/2012

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Overall project aims

Understand the role of poultry in transmitting bacteria creating a risk for consumers as a cause of foodborne illness and of resistance to antibiotic treatment

Map the pathways of contamination from production to consumption

Identify approaches to control contamination of poultry meat with bacteria causing foodborne illness and carrying antimicrobial resistance

Identify points of contamination and practices to prevent/ control it

Identify measures to control antibiotic use in poultry production and human medicine

Thailand Workshop 17-19/09/2012

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Specific aims

Human illness: Investigate the incidence of human enteric illness caused by pathogens that could be transmitted by chicken consumption when they are resistant to antimicrobials Investigate the practices used in medical clinics for antimicrobial treatment among sick children

Thailand Workshop 17-19/09/2012

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Specific aims continued

Poultry meat production: Describe the poultry value chain and the pathway from farm to retail markets Determine the level of chicken contamination and AMR Indentify key points where contamination is reduced, increased or remains the same and factors that could cause these changes

Thailand Workshop 17-19/09/2012

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Chronology of events

Submission of document to 3rd expert groups meeting : Oslo, Norway, June 2011  Decision made on implementation of AGISAR/WHO project in Cambodia/end June 2011 FAO/ early 2012, an integration manner, the AMR assessment risks along the poultry chain continuum 30/01/2012, seminar to launch the study, with stakeholders

Thailand Workshop 17-19/09/2012

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Poultry production

 Cambodia, poultry production is important by its contribution ( 26,000 T/year, source OIE, Cambodia)

Household food security Income

Human health risks associated to poultry consumption are mostly, associated with pathogens (Salmonella subspecies,

Campylobacter spp, E coli pathogen…)

As well as the increasing presence of antimicrobial resistance in pathogenic and commensal bacteria (E coli,

Enterococcus ) isolated in poultry products

Thailand Workshop 17-19/09/2012

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Project approach WHO/FAO

Type of sample ( FAO)

  • No. sample

Sample type Site Lab test to be carried out Analysis

Live bird fresh faecal droppings at production site

20x2x2 (2 type of production

  • Large scale production
  • Poultry backyard villager

Pooled sample Kien Svay district Kandal province

  • Campylobacter
  • Salmonella

And AMR

  • Quali. for Salmonella and

Campylobacter Cecae of birds at slaughter site/market 20x2x2 cecae Deum Kor or O Russey market

  • Campylobacter
  • Salmonella
  • E. coli

And AMR

  • Quali. for Salmonella

Quantitative for Campylobacter and E. coli Carcass after slaughter - neck skin 20x2x2 (WHO) Fresh Unchilled neck skin Deum Kor or Orussey market

  • Campylobacter
  • Salmonella
  • E. coli
  • VRE

And AMR

  • Quali. Salmonella and

Quantitative for Campylobacter , E Coli and Entercoccus VRE Rinse/wash water (if common for birds) to determine potential for cross- contamination of carcasses e.g. in bucket or basins 20x2x2 100mL Deum Kor or O Russey market

  • Campylobacter
  • Salmonella
  • E. coli

And AMR Qualitative for Salmonella Quantitative for Campylobacter and E coli Surface of chopping boards that could result in significant cross-contamination between birds; sample at the end of the day or after multiple birds in contact 20x2x2 swab 10cm2 area Deum Kor or O Russey market

  • Campylobacter
  • Salmonella
  • E. coli

And AMR Qualitative for Salmonella and quantitative for Campylobacter and E coli Total FAO 400 WHO Samples

  • No. samples

Sample type site Lab test to be carried out Analysis Carcasses after slaughter - neck skin 200 during FAO sampling ( 80 included within FAO sample) And 160 other s samples after finishing FAO project Fresh unchilled Neck skin Deum Kor or O Russey market

  • Campylobacter
  • Salmonella
  • E. coli
  • V RE

And AMR

  • Quali. Salmonella ,

Campylobacter , E Coli and Entercoccus VRE Total WHO 200 for total 360 AGISAR samples

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Market profile and poultry sellers

Two markets: Orussey (OR) and Deumkor (DK) in South PP area OR market, 1,200 shops (25-30 shop poultry sellers)

 Daily average weight sold/ 800-1200kg  Sometime, highest weight daily sold reached 2T

DK market, ~ 1000 shops (11 shops poultry sellers) These two markets owned by the government.

Thailand Workshop 17-19/09/2012

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Phnom Penh markets

Orussey market

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Sampling design

Each poultry sellers in each market will be randomly selected A total of 5 poultry neck skin samples and 2 on each of other poultry chains, minimum 11 samples per week from Phnom Penh OR and DK markets will be sampled

Thailand Workshop 17-19/09/2012

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Poultry farm

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Poultry backyard villagers

Thailand Workshop 17-19/09/2012

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Poultry slaughtering in the market

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Poultry sample collection

Storage post- slaughter Retail Slaughter Transport Farm Contact surfaces Water bath used to clean carcasses Feed

Water in tubs or buckets used to clean carcasses and the that may result in cross-contamination 100mL water Faecal droppings Pool 5 individual droppings for

  • ne sample

Caecae collect before bird is slaughtered Pool 10 caecae for one sample Common contact surfaces that may result in cross- contamination such as chopping blocks/boards/tables, ground Swab 10cm2 area Fresh unchilled carcasses for retail sale Collect neck skin

Samples for AGISAR

1 4 2 5 3

FAO specimen selected a long the poultry slaughter continuum

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Poultry slaughter chain continuum

Chopping board Slaughter Transport Retail Storage post- slaughter Water bath

Fresh unchilled carcasses for retail sale Chilled frozen carcasses

Thailand Workshop 17-19/09/2012

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Sampling site along the poultry slaughter continuum( AGISAR + FAO)

Chopping board Slaughter Transport Retail Storage post- slaughter Washing Water

Fresh carcasses for retail sale  Collect neck skin Chilled frozen carcasses- Collect neck skin Caecae before bird is slaughter Fecal dropping at large scale farm and bacyard poultry villagers

1 2 4 5 3

FAO specimen selected a long the poultry slaughter process

Specimen for AGISAR

Thailand Workshop 17-19/09/2012

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Sampling process for Cambodia dry season (February-July, 2012)

Thailand Workshop 17-19/09/2012

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Sampling process during the Cambodia rainy season (September/2012 to January 2013)

Thailand Workshop 17-19/09/2012

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Sample for FAO and AGISAR

 The sampling process for FAO start from February to July for 200 specimens and September 2012 to January 2013/ 2012 to obtain 400 specimens  Animal samples for AGISAR/WHO  1st-160th specimen for Salmonella,  1-237th specimen for Campylobacter  and 1-360th specimen for E. coli and Enterococcus  Human samples for AGISAR/WHO  1st- 750th diarrhea selected from Cambodia NP hospital

Thailand Workshop 17-19/09/2012

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Project outputs

Prevalence of Salmonella spp. and Campylobacter

  • spp. in retail poultry and isolates of these

pathogens from inpatient children with diarrhoea Levels of contamination with faecal indicator bacteria, E. coli and Enterococcus spp. in retail chicken AMR profile of the bacterial isolates

Thailand Workshop 17-19/09/2012

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Project outputs continued

Assessment of antimicrobial use in patients and poultry broiler production Stakeholder meetings and recommendations on

  • ptions for control of Salmonella and

Campylobacter food safety risks and AMR transmission Baseline for future policy developments

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Preliminary results

Thailand Workshop 17-19/09/2012

  • 76 Salmonella and 63 Campylobacter strains
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Preliminary results of human samples

 February – Septembre/2012 83/750 fresh fecal samples from NPH  Two Salmonella were identified  One Salmonella Typhimurium with ESBL profile  No Campylobacter spp have been isolated in human samples yet

Thailand Workshop 17-19/09/2012

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Antibio-resistance study

 Each of bacterial Gram negative isolate (Salmonella, and E. coli )

To 16 different antibiotic disks : AM, AMC, IPM, TMP, CF, CTX, CRO, CS, C, TE, SSS, STX, S, GM, NA, CIP

 Each Campylobacter spp

 To 3 different antibitic disks: E, TE, CIP

 By using diffusion method on MH agarose plate, CMI and diameter of inhibition zone

A

Thailand Workshop 17-19/09/2012

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AMR fragment study

Gene cat A, cmlA, sul1, sul2, aadA, strA, tet(A), tet(B), tet(C), tem, shv, oxa, dhfrI, dhfrII, CTX, CMY1, CMY2, GYRA

E coli and Salmonella MDR isolates, 18 primers, for gene fragments: To compare with reference sequences of 48 strains (DTU) Campylobacter MDR isolates, primers, for gene fragments:

Gene Peptidyl transferase,V region,23S rRNA gene for E, tet (O), and GYRA

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1

Faecal droppings Pool 10 individual droppings for

  • ne sample

From large scale farm and poultry backyard villagers

Faecal droppings

Thailand Workshop 17-19/09/2012

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2

Caecae collect before bird is slaughtered Pool 10 caecae for one sample

Caecae

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3

Fresh unchilled carcasses for retail sale Collect neck skin

Unchilled neck skin

Thailand Workshop 17-19/09/2012

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4

Contact surfaces

Surface of chopping board

Thailand Workshop 17-19/09/2012

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5

Water bath used to clean carcasses

Thailand Workshop 17-19/09/2012

Washing water

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If carcasses are chilled or frozen and held overnight collect neck skin as for retail samples Don’t use for the analyses

Chilled or frozen and held overnight

Thailand Workshop 17-19/09/2012

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Stakeholders

Institut Pasteur du Cambodge

  • Pr. Kruy Sun Lay,
  • Dr. Bertrand Guillard,
  • Dr. Arnaud Tarantola,

Ms Hem Sopheak,

  • Mr. Yith Vuthy
  • Pr. Vincent Deubel,

MAFF, NaVRI

  • Dr. Sorn San

Ministry of Health, CDC Department

  • Dr. Sok Touch

National Pediatric Hospital,

  • Pr. Chhour Y Meng

Phnom Penh Health department Dr Moul Chanta

WHO, Geneva

  • Dr. Awa Aidara-Kane

Denmark National Food Institute, Technical University of Denmark,

  • Dr. Rene Hendriksen

Norway Norwegian School of Veterinary Science,

  • Pr. Kari Grave

FAO

Nutrition and Consumer Protection Division

Vittorio Fattori, Renata Clarke

Animal Production and Health Division

Patrick Otto

Australia Brisbane, Queensland,

Food Safety Principles

Patricia Desmarchelier, FAO-Cambodia Allal,Lotfi

Thailand Workshop 17-19/09/2012

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THANK YOU FOR YOUR ATTENTION

Thailand Workshop 17-19/09/2012