Brent Gilpin, Lisa McCallum, Virginia Hope Institute of Environmental Science & Research Limited

Manaaki Tangata Taiao Hokiprotecting people and their environment through science

Specialist Science Solutions

PulseNet Asia Pacific – Using KAREN to help advance

international collaborations for analysis of disease causing

bacteria for both research and public health action.

Brent Gilpin, Lisa McCallum, Virginia Hope

Institute of Environmental Science & Research Limited

© ESR 2008

E. coli O157:H7

© ESR 2008

Western USA, 1993

726 cases identified4 deaths$160 million cost to company

E. coli O157:H7

© ESR 2008

0

10

20

30

40

50

60

70

1 8 15 22 29 36 43 50 57 64 71

Day of Outbreak

Nu

mb

er o

f C

ases outbreak

detected 1993Meat recall

1993 Western States E. coli O157 Outbreak

726 cases4 deaths

39 d

Slide courtesy Bala Swaminathan, CDC Atlanta

PFGEPulsed field gel electrophoresis

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 171 3 4 6 7

© ESR 2008

Loading into well

© ESR 2008

PFGE Pulsed Field Gel Electrophoresis

© ESR 2008

PulseNet in action

Pathogens isolated from faecal sample

E. coli O157, Listeria,

Salmonella, Campylobacter

PFGE DNA fingerprints compared

using electronic databases

PFGE analysis of all isolates

Initiate and guide epidemiological

investigations of outbreaksInterventions

Preventative measures

© ESR 2008

E. coli O157:H7 from Fresh Spinach

© ESR 2008

Many other examples…

© ESR 2008

S. Enteriditis• 12 May, 9:30 PM–1 AM

- Discussions with Costco led to local recall

- Nut-specific questionnaire developed

- E-mail bulletin to all states, CDC, FDA, Canada

• 13 May- PulseNet query finds matching cases elsewhere

- Traceback implicates Paramount; other brands

- National recall initiated

© ESR 2008

0

10

20

30

40

50

60

70

1 8 15 22 29 36 43 50 57 64 71

Day of Outbreak

Nu

mb

er o

f C

ases outbreak

detected 1993Meat recall

1993 Western States E. coli O157 Outbreak

726 cases4 deaths

39 d

Slide courtesy Bala Swaminathan, CDC Atlanta

2002 Colorado E. coli O157 Outbreak

0

10

20

30

40

50

60

70

1 8 15 22 29 36 43 50 57 64 71

Day of Outbreak

Nu

mb

er o

f C

as

es

outbreak detected

18 dWith

Pul

seN

et

Initial recall of 354,200pounds of ground beefExpanded to 18.6 million pounds38 cases from 11 states, 17 hospitalizations, one death

© ESR 2008

Austria, Belgium, Bulgaria, Czech Republic, Denmark, England, Finland, France, Germany, Greece, Hungary,

Iceland, Ireland, Italy, Luxembourg, Norway, Poland, Portugal, Romania, Scotland, Slovenia, Spain, Sweden, The

Netherlands28 public health labs, 17 veterinary labs

Uruguay, Chile, Argentina, Columbia, Mexico, Brazil

Vietnam, Thailand, Taiwan, The Philippines, the People’s Republic

of China, New Zealand, Malaysia, Korea, Japan, India, Hong Kong,

Bangladesh, and Australia

© ESR 2008

PulseNet Asia Pacific• Vietnam, Thailand, Taiwan, The

Philippines, the People’s Republic of China, New Zealand, Malaysia, Korea, Japan, India, Hong Kong, Bangladesh, and Australia

© ESR 2008

USDA product testing - E. coli O157:H7

Meat Processor A Meat Processor B

SupplierA

SupplierB

SupplierC

New Zealand

PFGE-XbaI

20.0

0

40.0

0

100.

00

150.

00

200.

00

250.

00

400.

00

500.

00

600.

00

800.

00

1500

PFGE-BlnI

20.0

0

40.0

0

100.

00

150.

00

200.

00

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00

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2000 Key

USDA_OB060047

USDA_OB060052

WA___9681

LabID

.

.

.

LabNoSampledDatePFGE-XbaI-pattern

EXHX01.1401

EXHX01.0074

EXHX01.1401

SourceType

.

.

.

SourceSite

.

.

.

TypeDetailsSurnameForenamesIsolationMethodProjectLocationPFGE-BlnI-pattern

EXHA26.0569

EXHA26.0569

EXHA26.0569

AB

© ESR 2008

NZ beef exports to USA

• USA 51% of total NZ beef exports• Value of approximately 1 billion dollars…

© ESR 2008

Findings• 12 human, 3 meat isolates same as US XbaI 74

- (This XbaI 5.9% of NZ isolates)

• All of have BlnI patterns different to US BlnI 569• No isolates with US XbaI pattern 1401• Two isolates with BlnI pattern 569 (different Xba)• No evidence to suggest isolates came from NZ

meat

PFGE-XbaI

20.0

0

40.0

0

100.

00

150.

00

200.

00

250.

00

400.

00

500.

00

600.

00

800.

00

1500

PFGE-BlnI

20.0

0

40.0

0

100.

00

150.

00

200.

00

300.

00

400.

00

500.

00

600.

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1000

2000 Key

USDA_OB060047

USDA_OB060052

WA___9681

LabID

.

.

.

LabNoSampledDatePFGE-XbaI-pattern

EXHX01.1401

EXHX01.0074

EXHX01.1401

SourceType

.

.

.

SourceSite

.

.

.

TypeDetailsSurnameForenamesIsolationMethodProjectLocationPFGE-BlnI-pattern

EXHA26.0569

EXHA26.0569

EXHA26.0569

© ESR 2008

Key components• Standardised methodology for lab analysis• Computer software analyse data• Databases of patterns• Communication among participants

- Outbreaks

- Education and upskilling

© ESR 2008

Role of KAREN network• Communication among participants

- Outbreak investigations

- Education

• Regional and international databases

© ESR 2008

IDC

Asia-PacificDatabase Server

New Zealand

E. coli O157 Listeria Salmonella Shigella Neisseria Vibrio Campylobacter

PWR

OK

WIC0ACT/CH0

ACT/CH1

WIC0ACT/CH0

ACT/CH1

ETHACT

COL VPN

Japan China Hong Kong India Taiwan Bangladesh New Zealand

IDC

CDC Database ServerAtlanta, GA, USA

IDC

Canadian DatabaseServer

Winnipeg, MB, Canada

Asia-Pacific BackupDatabase Server

New Zealand

IDC

IDC

National DatabaseServer

Option 1

Hardware/SoftwareDatabase managers

Regional Databases

© ESR 2008

New ZealandMain Hub

IDC

IDC

IDC

IDC

IDC

IDC

IDC

IDC

JapanE. coli O157

ChinaListeria

Hong KongSalmonella

IndiaShigella

TaiwanNeisseria

BangladeshVibrio

New ZealandCampylobacter

Data CopyData Copy

Data Copy

Data Copy

Data Copy Data Copy

Data Copy

IDC

IDC

CDC NML

IDC

IDC

IDC

IDC

IDC

IDC

IDC

IDC

Backup Server

Backup Server

Backup Server

Backup Server

Backup Server

Backup Server

Backup Server

Backup Server

Option 2Regional Databases

© ESR 2008

Export data wish to submit(XML)

Upload to folder on PNAP forum

Participants

Database coordinator

Download new submission

1. Check “compatibility”2. Incorporate into

“Regional database”3. Post updated

database into PNAP forum folder

Download updated database

Participants

Open regional database as “local copy” using BioNumerics

• Very simple solution• Every country equally important• Ensures access and sharing of data• Allows distribution of work, lower commitment required

from coordinating lab.• Benefits

- Archive of genotypes

- Evaluations of variability in genotype

- Geographical and temporal spread of genotypes

- Get clues about potential sources

- Evaluate how common/rare a genotype is

© ESR 2008

PulseNet Asia Pacific portal• PNAP portal places NZ as the coordinating

hub for the region.• Communication forum • Provides a mechanism for regional

databases for Asia Pacific• Would like capacity for video meetings of

participants from Asia Pacific, and also PulseNet International.

• Would like capacity for educational sessions on both laboratory and computer analysis, and for research and outbreak seminars.

© ESR 2008

• Connect New Zealand’s highest level bio-containment laboratory at Wallaceville to the rest of NZ and the world via video microscopy and video conferencing.

National Centre for Biosecurityand Infectious Disease - Wallaceville

Brent Gilpin, Lisa McCallum, Virginia Hope Institute of Environmental Science & Research Limited

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