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Marianne Sunde National Veterinary Institute Oslo, Norway

Monitoring antimicrobial resistance in the veterinary field: Norway -Special reference to MRSA, ESBLs and antimicrobial use to farmed fish . Marianne Sunde National Veterinary Institute Oslo, Norway. Outline:. The Norwegian monitoring programme for antimicrobial resistance

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Marianne Sunde National Veterinary Institute Oslo, Norway

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  1. Monitoring antimicrobial resistance in the veterinary field: Norway-Special reference to MRSA, ESBLs and antimicrobial use to farmed fish Marianne Sunde National Veterinary Institute Oslo, Norway

  2. Outline: • The Norwegian monitoring programme for antimicrobial resistance • MRSA from animals in Norway • ESBLs from animals in Norway • Antimicrobial usage to farmed fish • Methicillin resistant Staphylococcus pseudintermedius (MRSP) from dogs in Norway

  3. NORM-VET monitoring programme for antimicrobial resistance in the veterinary and food production sectors • Running since year 2000 • Annual joint report NORM (human)/NORM-VET • NORM-VET resistance testing of: - zoonotic pathogens – Campylobacter-Salmonella - indicator bacteria – E. coli, Enterococci - animal pathogens - E. coli, Staphs, Ent.hirae - indicator bacteria from wild animals – E. coli - fish pathogens – Moritella viscosa, Vibrio anguillarum

  4. Bacterial isolates to NORM-VET: • From other surveillance and control programs • From veterinarians/clinics invited to participate • Bacteriological diagnostic service – National Veterinary Institute • Methods: Broth micro dilution method (VetMICTM) • NORM-VET is coordinated by the Norwegian Zoonosis Centre Resistance testing: Section of bacteriology National Veterinary Institute, Oslo

  5. Resistance trends during 2000-2010 • Food producing animals - indicator bacteria; Relatively low/moderate resistance occurrence (E. coli) - resistance to streptomycin, tetracycline, sulphonamides, ampicillin - animal species variation: lowest occurrence from sheep and cattle, higher in pigs and broiler Relatively stable resistance frequencies, BUT: - increase in quinolone resistance in E. coli from broiler in 2009 ?? • Food producing animals - pathogens; - For many bacterial species limited samples sizes - Relatively low resistance occurrence, example: S. aureus mastitis 5-7% PENR

  6. Resistance trends during 2000-2010 • Zoonotic pathogenic bacteria: - Relatively low/moderate resistance occurrence - Example: Campylobacter jejuni broiler: less than 5% Cipr/NalR Salmonella: Low prevalence in animals in Norway Low resistance occurrence Salmonella reservoar in Norway: Wild birds and hedgehog – susceptible variants

  7. Resistance trends during 2000-2010 Companion animals: -Resistance among β-hemolysin producing Staphylococci from dogs: PENR 75%, TETR 40%, FUSR 60% - Increased MRSP occurrence in Norway 2008-2010

  8. NORM-VET – future challenges: Reduced program ? - year 2000/2001: 13-1400 isolates - year 2008/2009: 6-800 isolates More use of selective methods (MRSA – ESBL) More data on fish pathogens ?

  9. MRSA in animals - Norway • MRSA ST398 not detected from animals in Norway • Have we searched for MRSA in animals ? Baseline survey 2008, dust from 256 pig holdings, one MRSA finding, MRSA ST8/t008 Slaughterhouse study in 2008, 1000 pigs (nasal swabs) 200 holdings, no MRSA, 2 S. aureus Horses NORM-VET 2009, 200 horses, nasal swabs – no MRSA Pigs in NORM-VET 2011 ??

  10. MRSA ST8/t008 in a pig holding • MRSA ST8/t008; a common human MRSA type in Norway • Low prevalence of MRSA among humans in Norway Most frequent spa-types in Norway in 2008 Norwegian reference laboratory for MRSA EARSS Annual Report 2006. Bilthoven: European Antimicrobial Resistance Surveillance System (EARSS), 2007

  11. MRSA ST8/t008 in a pig holding • Most probably human-to-animal transmission of MRSA • Human to livestock MRSA transmission • The pigs were not heavily colonized with MRSA – in contrast to what is reported for MRSA ST398 in pigs • MRSA ST8/t008 perhaps more adapted to humans ? Not the same ability to colonize/persist as animal(pig) adapted S. aureus types ?

  12. MRSA in animals in Norway • Other known animal MRSA cases in Norway: Year Species Source ST/CC/spatype 2002 Horse Unknown ST8 2008 Cat MRSA positive owner ST45 t1081 2008 Dog MRSA positive owner ST8 t324 2008 Dog Surgery in Spain ? ST22 t032 2009 Cat MRSA positive owner ST8 t008

  13. ESBLs in animals - Norway • Food producing animals One isolate, E. coli broiler, NORM-VET 2006 • Companion animals Six isolates (three ESBLs/three AmpC), all E. coli One isolate from NORM-VET 2008 Five from diagnostic submissions, National Veterinary Institute

  14. ESBLs in animals - Norway • ESBL from broiler, NORM-VET 2006 • Intestinal flora of healthy animal • Low MICs to cephalosporins: MIC=1 MIC=4

  15. ESBLs in animals - Norway Genetic investigasjon ESBL broiler: • blaTEM-20 gene on a conjugative plasmid • Plasmid replicon typing: incI1 plasmid • blaTEM-20 gene with similar MICs from in Salmonella paratyphi B dT+ from poultry in the Netherlands (Hasman 2005) • Comparison studies: • Both blaTEM-20 genes located on incI1 plasmids • blaTEM-20 nt sequence and promoter 100% identical → Equal plasmids in E. coli from broiler in Norway and Salmonella paratyphi B+ from the Netherlands ?

  16. ESBLs in animals - Norway • Plasmid multi locus sequence typing (pMLST) of incI1 plasmids (Garcia-Fernandez et al 2008) • Amplification and sequencing of five regions on incI1 plasmids, allelic profil • Result: Identical nt sequences both incI1 plasmids → equal/closely related blaTEM-20 plasmids from Norway and the Netherlands (Sunde et al, 2009) Why ESBLs in broiler in Norway ? - No selection pressure for cephalosporin usage - Import of breeding animals to Norway ? - A ”fitness” plasmid ? ?

  17. ESBLs in animals - Norway • One nt difference between blaTEM-20 and blaTEM-52 • blaTEM-52 mediates high level resistance to cephalosporins • A incI1 plasmid carrying blaTEM-52 is widely disseminated in Salmonella from humans and poultry in France and Belgium (Clockart et el 2007)

  18. ESBLs in animals - Norway • Sequencing of the resistance region (6 KB) → blaTEM-20 and blaTEM-52 the same genetic organization • pMLST showed that the blaTEM-20 and blaTEM-52 plasmids are closely related (Sunde et al, 2009)

  19. ESBLs in animals - Norway • Overview of ESBLs and AMPc from animals in Norway: year Species gene 2006 broiler, healthy blaTEM-20 2007 dog, wound CTX-M-15 2008 dog, healthy CTX-M-15 2009 dog, infection CTX-M-14 2009 dog, wound postoperative blaCMY 2009 dog, sepsis puppy blaDHA 2010 dog, skin infection blaCMY

  20. Antimicrobial agents to farmed fish - Norway Intensive fish farming since mid 1970s - Atlantic salmon - main species Succeed in domestication of salmon has been of major importance Other species: Cod, Halibut, Trout

  21. Antimicrobial agents to farmed fish - Norway • Major diseases today: • Viral diseases (pancreas disease/infectious pancreas necrosis) • Bacterial infections (Moritella viscosa, Francisella spp) Antimicrobial agents to farmed fish (NORM/NORM-VET 2009):

  22. Antimicrobial agents to farmed fish - Norway Disease control – a vital factor for the expansion of the aquaculture industry Problems with infectious diseases during the 80s and early 90s: • Vibrio anguillarum (cold-water vibriosis) • Vibrio salmonicida • Aeromonas salmonicida (furunculosis) → Gave mass mortality and massive antibiotic consumption Today: controlled by effective vaccines and preventive measures

  23. Antimicrobial agents to farmed fish - Norway

  24. Antimicrobial agents to farmed fish - Norway Grave and Hanssen 2005 The antimicrobial usage to farmed fish is reduced by 98% during the last 20 years

  25. Antimicrobial agents to farmed fish – future challenges • Domestication and farming of new species – new diseases • Farmed cod – Francisella spp infections salmon 2008 Ca. 741 000 tons active- ingredient (kg) cod 2008 Ca. 13 500 tons Norwegian Institute of Public Health cod 65%

  26. Methicillinresistant Staphylococcus pseudintermedius (MRSP) from dogs • First case 2008 – outbreak small animal clinic • Increase in MRSP from dogs ? - 2008: MRSP from 2 dogs - 2009: MRSP from 7 dogs - 2010: MRSP from 12 dogs • Molecular typing (MLST - PFGE): - Clonal diversity - Few isolates belonging to ST 71 (the dominant clone in Europe ?)

  27. Antimicrobial resistance in Norway – veterinary sector • Food producing animals: • relatively low/moderate occurrence of resistance • but: MRSA and ESBL detected • Companion animals: • relatively high resistance prevalences (Staphylococci) • MRSA and ESBLs detected • MRSP not uncommon ? • Farmed fish: • limited/no knowledge resistance in fish pathogens • usage of antimicrobial agents stable • but: Farmed cod – increased usage ??

  28. Thank you for your attention

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