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 %
t002
t008
t019
29,1 %
t044
10,1 %
t304
t223
t437
t032
t067
t127
9,6 %
nt
t010
1,3 %
1,3 %
1,3 %
1,3 %
1,4 %
1,4 %
1,4 %
1,4 %
1,9 %
2,0 %
2,2 %
t015
t026
t690
6,8 %
t021
t065
5,4 %
2,6 %
2,9 %
4,8 %
t311
t324
Others
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
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
2002
2008
2008
2008
2009
Species
Horse
Cat
Dog
Dog
Cat
Source
Unknown
MRSA positive owner
MRSA positive owner
Surgery in Spain ?
MRSA positive owner
ST/CC/spatype
ST8
ST45 t1081
ST8 t324
ST22 t032
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=1
MIC=4
ESBLs in animals - Norway
Genetic investigasjon ESBL broiler:



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

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)
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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 - Norway
One
nt difference between blaTEM-20 and blaTEM-52
blaTEM-52 mediates
A
high level resistance to cephalosporins
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
2006
2007
2008
2009
2009
2009
2010
Species
gene
broiler, healthy
blaTEM-20
dog, wound
CTX-M-15
dog, healthy
CTX-M-15
dog, infection
CTX-M-14
dog, wound postoperative blaCMY
dog, sepsis puppy
blaDHA
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
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
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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 - 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
Antimicrobial agents to farmed fish - Norway
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
Antimicrobial agents to farmed fish – future challenges


Domestication and farming of new species – new diseases
Farmed cod – Francisella spp infections
1600
1400
salmon 2008
Ca. 741 000
tons
1200
1000
active800
ingredient
600
(kg)
cod 2008
Ca. 13 500
tons
400
200
0
2001
2002
2003
2004
2005
2006
2007
2008
Norwegian Institute of Public Health
cod 65%
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 ??
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Thank you for your attention