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

MRSA from animals in Norway

ESBLs from animals in Norway

Antimicrobial usage to farmed fish

Methicillin resistant Staphylococcus pseudintermedius (MRSP) from dogs in Norway

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

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

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

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

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

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 ?

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 ??

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

11,7 %

10,1 %

9,6 %

6,8 %

5,4 %

4,8 %2,9 %2,6 %2,2 %2,0 %

1,9 %1,4 %

1,4 %1,4 %

1,4 %1,3 %

1,3 %1,3 %

1,3 %

29,1 %

t002

t008

t019

t044

t304

t223

t437

t032

t067

t127

nt

t010

t015

t026

t690

t021

t065

t311

t324

Others

Most frequent spa-types in Norway in 2008Norwegian reference laboratory for MRSA

EARSS Annual Report 2006. Bilthoven: European Antimicrobial Resistance Surveillance System (EARSS), 2007

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 ?

MRSA in animals in Norway

Other known animal MRSA cases in Norway:

Year Species Source ST/CC/spatype

2002 Horse Unknown ST82008 Cat MRSA positive owner

ST45 t10812008 Dog MRSA positive owner ST8

t3242008 Dog Surgery in Spain ?

ST22 t0322009 Cat MRSA positive owner ST8

t008

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

ESBLs in animals - Norway

ESBL from broiler, NORM-VET 2006 Intestinal flora of healthy animal Low MICs to cephalosporins:

MIC=1MIC=4

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 ?

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 ? ?

ESBLs in animals - NorwayOne 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)

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)

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

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

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):

Antimicrobial agents to farmed fish - NorwayDisease control – a vital factor for the expansion of the aquaculture

industryProblems 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

Antimicrobial agents to farmed fish - Norway

Antimicrobial agents to farmed fish - Norway

The antimicrobial usage to farmed fish is reduced by 98% during the last 20 years

Grave and Hanssen 2005

Antimicrobial agents to farmed fish – future challenges Domestication and farming of new species – new

diseases Farmed cod – Francisella spp infections

0

200

400

600

800

1000

1200

1400

1600

2001 2002 2003 2004 2005 2006 2007 2008

active-ingredient (kg)

cod 65%

salmon 2008Ca. 741 000 tons

cod 2008Ca. 13 500 tons

Norwegian Institute of Public Health

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 ?)

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 ??

Thank you for your attention