risk-based food safety inspection during incident event in...

___________________________________________________________________________

2014/SCSC/WKSP1/008

Risk-Based Food Safety Inspection During Incident Event in Thailand

Submitted by: Thailand

Workshop on Improved Food Inspection Capacity Building Based on Risk Analysis

Seoul, Korea21-23 May 2014

Risk-Based Food Safety Inspection during Incident Event in Thailand

Dr. Waraluk Tangkanakul

Acting Deputy Director, Food Safety Extension and Support Bureau, Ministry of Public Health, Thailand

Content

• Investigation of a Multi-provinces Cholera Outbreak in Thailand which expanded to Laos People Democratic Republic in 2007

• Investigation and develop risk-based inspection (RBI) along exported Cockles' food supply chain (from Thailand across several check points to Laos PDR)

• Future RBI during incident event uses Government Information Network (GIN) system


• Investigation and develop risk-based inspection (RBI) along exported Cockles' food supply chain (from Thailand across several check points to Laos PDR)


q 988 cases with 7 death (CFR 0.7%) q spread to 50 provinces of total 76 provinces q Inaba: most occur in Tak province q Ogawa: distribute for the whole country q age group 15-44 years

Number

Lao

Lumphun

TakKhonkaen

Situation of Cholera in Thailand, Situation of Cholera in Thailand, 20072007

Age group

0

50

100

150

200

250

<1 เดือน 1เดือน-4 ปี 5-9 ปี 10-14 ปี 15-24 ปี 25-34 ปี 35-44 ปี 45-54 ปี 55-64 ปี >65 ปี

Number

Cockle farm

Hospitalized 926 Community search 433 Death 7 Geographical distribution 50 provincesOgawa 1,087 Inaba 266 Hikojima 1

O139 5

Ogawa

Cholera cases by Cholera cases by V. V. choleraecholerae serotype in Thailand, serotype in Thailand, 20072007

Hospitalized 926 Community search 433 Death 7 Geographical distribution 50 provincesOgawa 1,087 Inaba 266 Hikojima 1

O139 5

Inaba38th

In 2007: Cholera outbreak

Thailand 10 Oct. 2007

( 38thwk), expanded to Laos People

Democratic Republic

MultiMulti--provinces Cholera provinces Cholera outbreak associated with cocklesoutbreak associated with cockles

Most V. cholerae strains isolated from patients “V. cholerae O1 serotype Ogawa”


5



The cockles were assumed to be a source of cholera outbreak (OR 2.24, 95% CI 1.43 – 4.02)

The cockles were assumed to be a source of cholera outbreak (OR 2.24, 95% CI 1.43 – 4.02)

Vientiane, Laos PDR Thasadet

Check point, Nong Khai, Thailand

ThaiThai--Laos ‘s Joint investigation on Cholera outbreak, Laos ‘s Joint investigation on Cholera outbreak, 20072007

5th Oct. 2007: Laos,male27 years working in a paper factory in VientianeSuspected food : parboil cockles (half kg) Rx: Doxycycline at Mahosot hospital, Laos Organism: RSC positive for Vibrio cholerae O1 El Tor Ogawa (10th Oct.)Investigation : 120 workers in factory (80 living), 10 RSC sample found 1 positive the same organism (case’s 2 months pregnant wife)




Content


• Investigation and develop risk-based inspection (RBI) along exported cockcles’ food supply chain (from Thailand across several check points to Laos PDR)



• Investigation and develop risk-based inspection (RBI) along exported cockcles’ food supply chain (from Thailand across several check points to Laos PDR)


Scientific classification

Kingdom: Animalia

Phylum: Mollusca

Cockles: Cockles: AnadaraAnadara granosagranosa

: marine animal which is habitat of marine microorganisms

8

Phylum: Mollusca

Class: Bivalvia

Subclass: Pteriomorphia

Order: Arcoida

Family: Arcidae

Genus: Anadara

Species: A. granosa

In Thailand,cockle is usually cooked by boiled or roasted

transmitted the pathogensto human via contaminated cockle

Investigation of cockles’ food supply chain from Thailand to Laos PDR

Transport to Northeastern Provinces of Thailand

Pass out at 4 check points (Thailand) and enter at 2 check points (Laos PDR)

Farm in Surat Thani

Transport to Northeastern Provinces of Thailand

Inspection of cockles’ farm

• Departments of FisheriesCoastal Fisheries Research and Development SuratThani: care, promote and monitor of water quality affecting cockle farming. The amount of salinity of sea water should be 10-30 ppt.

• Fisheries Office Surat Thani : 1. Promote the coastal fishery and Good Aquaculture

Standard (GAP) and registration cockles’ farm.2. Research and monitor of water quality and aquatic

products from Surat Thani.3. Check for contamination and quality of aquatic

products.

• Departments of FisheriesCoastal Fisheries Research and Development SuratThani: care, promote and monitor of water quality affecting cockle farming. The amount of salinity of sea water should be 10-30 ppt.

• Fisheries Office Surat Thani : 1. Promote the coastal fishery and Good Aquaculture

Standard (GAP) and registration cockles’ farm.2. Research and monitor of water quality and aquatic

products from Surat Thani.3. Check for contamination and quality of aquatic

products.

In 2007, the salinity of sea water was less than 10 ppt. for long and continuous months (Aug. – Oct.)

Salinity of sea water in cockles’ farm

11

In 2007, the salinity of sea water was less than 10 ppt. for long and continuous months (Aug. – Oct.)

In 2007, total rainfall in SuratThani was more than 90.1 millimeter.

Total rainfall in Surat Thani in 2007

12

In 2007, total rainfall in SuratThani was more than 90.1 millimeter.

Food quality inspection for V.cholerae

13

Conclusion of Cholera Investigation (1)

• The outbreak of cholera in Laos – Thailand linked together because the investigation revealed that the suspect food was exported -imported cockles which found V. cholerae

• Decreasing of sea water’s salinity in 2007 which promoted the high prevalence of V. cholerae might be because of heavy rain fall.

• The outbreak of cholera in Laos – Thailand linked together because the investigation revealed that the suspect food was exported -imported cockles which found V. cholerae

• Decreasing of sea water’s salinity in 2007 which promoted the high prevalence of V. cholerae might be because of heavy rain fall.

Conclusion of Cholera Investigation (2)

• Transportation time of cockles from SuratThani through Loas PDR was greater than 20 hours may be an important factor in inducing the increasing number of V. cholerae until reaching infective dose (109).

• To prove the hypothesis, pulse field gel electrophoresis should be used to identified Vibrio cholerae strain caused this outbreak was the same or not ?

• GAP should be strengthening by regulatory agencies.

• Transportation time of cockles from SuratThani through Loas PDR was greater than 20 hours may be an important factor in inducing the increasing number of V. cholerae until reaching infective dose (109).

• To prove the hypothesis, pulse field gel electrophoresis should be used to identified Vibrio cholerae strain caused this outbreak was the same or not ?

• GAP should be strengthening by regulatory agencies.

Cockles samples Rectal swab samples- Patients, family

members, neightboring

Detection of Vibrio cholerae from specimens by culture method

(N= 161)

(N=328)

RBI for cholera outbreak during 2007 to 2010, Thailand

Detection of Vibrio cholerae from specimens by culture method

Detection of Vibrio cholerae serogroups by Uniplex and Duplex PCR, PFGE, Ribotyping , MLVA techniques.

V. cholerae non-O1V. cholerae O1

PCR : Polymerase Chain Reaction, PFGE : Pulsed field gel electrophoresisMLVA : Ribotyping, multiple-locus variable-number tandem-repeat analysis

Sample collection1. Khon Kaen province : 30 samples2. Udon Thani provice : 7 samples3. Nong Khai province : 56 samples4. Exported cockles: 68 Samples (sample

from truck (once/ month from Mar. to Nov. 2010)

Study method1.Standard culture method2. PCR




Risk based inspection for cockles, 2010

17

Obj: To define the prevalence of V. cholerae in cockles

Obj: To define the prevalence of V. cholerae in cockles







Detection of Vibrio spp. in cockles from 3- sources of collection

250 ml. of PBS250 g of cockle

250 g of cockle suspended in 250 ml of PBS

Added 20 ml (10 g) of cockle suspension to 80 ml of APWAdded 20 ml (10 g) of cockle suspension to 80 ml of APW

18

Added 20 ml (10 g) of cockle suspension to 80 ml of APWAdded 20 ml (10 g) of cockle suspension to 80 ml of APW

APWAPW

Incubated at 37oC for 6 hIncubated at 37oC for 6 h

A loopfulA loopful1 ml1 ml

Culture method Culture method PCR methodPCR method

DNA extractionDNA extraction

PCR amplificationPCR amplification

Streak on TCBS agarStreak on TCBS agar

Biochemical testBiochemical test

Prevalence of Vibrio spp. in 143 cockles detected by tetraplexPCR, uniplex PCR and culture method

72

10091.6

9.8

81.8

10095.1

23.1

0.7

79.7

37.1

2.1

0

20

40

60

80

100

V.cholerae V. parahaemolyticus V. vulnificus V. mimicus

Nu

mb

er o

f po

siti

ve s

amp

les

(%)

Tetraplex PCR

Uniplex PCR

Culture

19

72

10091.6

9.8

81.8

10095.1

23.1

0.7

79.7

37.1

2.1

0

20

40

60

80

100


Nu

mb

er o

f po

siti

ve s

amp

les

(%)

Tetraplex PCR

Uniplex PCR

Culture

Fig. Prevalence of Vibrio spp. in 143 cockles detected by tetraplex PCR, uniplex PCR and culture methodThe prevalence of Vibrio spp. detected by tetraplex PCR was slightly lower

than uniplex PCR however, it higher than culture methodThe prevalence of Vibrio spp. detected by tetraplex PCR was slightly lower

than uniplex PCR however, it higher than culture method

Prevalence of Vibrio spp. in three different sources of cockle samples (uniplex PCR)

85.3

10091.2

17.6

80.4

100 96.4

3.6

81.1

100 96.2

47.2

0

20

40

60

80

100


Num

ber o

f pos

itive

sam

ples

(%

)

Exported to Lao PDR (34)

Nong Khai (56)

Khon Kaen(53)

20

85.3

10091.2

17.6

80.4

100 96.4

3.6

81.1

100 96.2

47.2

0

20

40

60

80

100


Num

ber o

f pos

itive

sam

ples

(%

)

Exported to Lao PDR (34)

Nong Khai (56)

Khon Kaen(53)

Fig. Prevalence of Vibrio spp. in three different sources of cockles samples§ Vibrio spp. among three sources were not significantly different, except V. mimicus was found very high (47.2%) in Khon Kaen

§ V. parahaemolyticus was found in 100% in all 3 sources§ V. vulnificus was found about 90% in all 3 sources§ V. cholerae was found about 80% in all 3 sources

§ Vibrio spp. among three sources were not significantly different, except V. mimicus was found very high (47.2%) in Khon Kaen

§ V. parahaemolyticus was found in 100% in all 3 sources§ V. vulnificus was found about 90% in all 3 sources§ V. cholerae was found about 80% in all 3 sources

Mixed Vibrio spp. found in cockle samples in the northeastern Thailand

14.7

58.8

11.8 14.7

03.6

75

1.8

19.6

0

37.741.5

0

17

3.8

18.9

58.7

3.5

17.5

1.4

0

20

40

60

80

100

VP+VC+VV+VM VP+VC+VV VP+VC VP+VV VP

Num

ber o

f pos

itiv

e sa

mpl

es (%

) Exported to Lao PDR (34)

Nong Khai (56)

Khon Kaen(53)

Total

Fig. Mixed Vibrio spp. found in cockle samples in the northeastern Thailand

21

14.7

58.8

11.8 14.7

03.6

75

1.8

19.6

0

37.741.5

0

17

3.8

18.9

58.7

3.5

17.5

1.4

0

20

40

60

80

100

VP+VC+VV+VM VP+VC+VV VP+VC VP+VV VP

Num

ber o

f pos

itiv

e sa

mpl

es (%

) Exported to Lao PDR (34)

Nong Khai (56)

Khon Kaen(53)

Total

VC : V. cholerae, VV : V. vulnificus, VP: V. parahaemolyticus, VM : V. mimicusVC : V. cholerae, VV : V. vulnificus, VP: V. parahaemolyticus, VM : V. mimicus

All of cockle samples were found at least one Vibrio spp. and most of them were found more than two Vibrio spp. (77.6%)

All of cockle samples were found at least one Vibrio spp. and most of them were found more than two Vibrio spp. (77.6%)Most of cockle samples were contaminated with combination of

various species of Vibrio.Most of cockle samples were contaminated with combination of

various species of Vibrio.

Summary of the sensitivity of the tetraplex PCR and uniplex PCR compared with culture method using spiked Vibrio spp. in sterile cockle samples

Pathogens Tetraplex PCR

0 h 3 h 6 h 18 h

V. cholerae - 1 1 1

V. parahaemolyticus - 101 1 1

Uniplex PCR

0 h 3 h 6 h 18 h

- 1 1 1

- 1 1 1

Culture method

0 h 3 h 6 h 18 h

- 101 1 1

- 102 101 1

22

V. parahaemolyticus - 101 1 1

V. vulnificus - 102 1 1

V. mimicus - 102 102 102

- 1 1 1

- 101 1 1

- 101 101 101

- 102 101 1

- 102 101 1

- 103 102 102

After 6 h of enrichment in APW is sufficient for detection of Vibrio spp. in cockle samples

because as few as 1 CFU can be detected by tetraplex PCR

According to the DMSC food standard, the cockles distributing in the market in Nong Khai and Khon Kaen, and exported to lao PDR were not up to standard due to almost of

cockle samples were presence of V. parahaemolyticus and V. cholerae in 10 g of cockle sample

whereas the criteria for V. parahaemolyticus and V. cholerae require has to be absent in 25 g of food sample

(frozen and chilled fishery product)





Food standard

23









Conclusion RBI on cockles, 2010

• V. cholerae was detected only by PCR method.• Enrichment at least 6 hr was increased the

detection.• The reason of low prevalence of V. cholerae by

culture method because of it might be in the stage of viable but non-culturable (VBNC)

• The results showed the important of cockles to spread V. cholerae and demonstrated PCR should be used for RBI

• V. cholerae was detected only by PCR method.• Enrichment at least 6 hr was increased the

detection.• The reason of low prevalence of V. cholerae by

culture method because of it might be in the stage of viable but non-culturable (VBNC)

• The results showed the important of cockles to spread V. cholerae and demonstrated PCR should be used for RBI

Conclusion RBI on human samples, 2007 -2010, Thailand (1)

• The cholera outbreak in Thailand during 2007 – 2010 were exclusively caused by the V. cholerae O1 El Tor variant carrying the classical ctxB and El Tor rstR genes. It probably appeared in Thailand during recent years.

• PFGE differentiated Thai El Tor variant isolates into nine pulsotypes that share the similarity of 88%.

• The cholera outbreak in Thailand during 2007 – 2010 were exclusively caused by the V. cholerae O1 El Tor variant carrying the classical ctxB and El Tor rstR genes. It probably appeared in Thailand during recent years.

• PFGE differentiated Thai El Tor variant isolates into nine pulsotypes that share the similarity of 88%.

Figure 1. Pulsed-field gel electrophoresis (PFGE) patterns among 343 Thai V. cholerae O1 isolates.

Okada K, Roobthaisong A, Nakagawa I, Hamada S, et al. (2012) Genotypic and PFGE/MLVA Analyses of Vibrio cholerae O1: Geographical Spread and Temporal Changes during the 2007–2010 Cholera Outbreaks in Thailand. PLoS ONE 7(1): e30863. doi:10.1371/journal.pone.0030863http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030863

Conclusion RBI on human samples, 2007 -2010, Thailand (2)

• MLVA typing among isolates during outbreak episodes in different geographical and association of causative V. cholerae in cholera outbreak showed the different of MLVA typing.. In 2007, in northeastern region were triggered by the consumption of cockles contaminated with V. cholerae MLVA type 2.

• The study can be the data to trace the emergence, year long survival , or disappearance of a particular type (s) of isolate in terms of spatial and temporal association

• MLVA typing among isolates during outbreak episodes in different geographical and association of causative V. cholerae in cholera outbreak showed the different of MLVA typing.. In 2007, in northeastern region were triggered by the consumption of cockles contaminated with V. cholerae MLVA type 2.

• The study can be the data to trace the emergence, year long survival , or disappearance of a particular type (s) of isolate in terms of spatial and temporal association

Figure 3. Distribution of major MLVA types of V. cholerae O1 isolates during the 2007–2010 cholera outbreaks in Thailand.

Okada K, Roobthaisong A, Nakagawa I, Hamada S, et al. (2012) Genotypic and PFGE/MLVA Analyses of Vibrio cholerae O1: Geographical Spread and Temporal Changes during the 2007–2010 Cholera Outbreaks in Thailand. PLoS ONE 7(1): e30863. doi:10.1371/journal.pone.0030863http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030863

รอ้ยละ

100

67.9

85.781.8

98.2 100

80

100

120

29

50

18.232.1

50

14.3

1.8

0

20

40

60

C Nor Gm Amp E Te Sxt

Green: sensitive, light orange = intermediate, thick orange = resistance

Content


• Investigation and develop risk-based inspection (RBI) along exported Cockcles food supply chain (from Thailand across several check points to Laos PDR)



• Investigation and develop risk-based inspection (RBI) along exported Cockcles food supply chain (from Thailand across several check points to Laos PDR)


GIN system

8.00

10.00

12.00

14.00

12.78

5.81

7.47

8.58

6.11

ร้อยล

ะทีไม่ผ่าน

เกณฑ์

ปีงบประมาณ

2546

2547

2548

0.00

2.00

4.00

6.00

8.00

สารเร่งเนือแดง บอแรกซ์ สารฟอกขาว ฟอร์มาลนี สารกนัรา ยาฆ่าแมลง

1.77

5.81

3.02

5.56

7.47

0.84

2.00

1.92

4.74 5.4

46.11

0.54

0.37 1.1

1 1.35

3.98

2.17

1.50

0.51

0.57

0.01

3.14

1.63

2.82

0.40

0.38

0.21

2.23

0.83

0.24

0.09 0.3

3

0.27

1.55

0.17

0.17

0.01 0.3

8

0.25

2.27

0.82

0.52

0.52 1.3

6

0.72

2.97

1.70

0.09

0.01 0.6

8

0.25

1.59

1.50

0.62

0.08 0.6

9

0.37

2.83ร้อ

ยละท

ีไม่ผ่าน

เกณฑ์

2549

2550

2551

2552

2553

2554

2555

Beta agonist, Borax, Sodium hydrosulfite, Formalin, anti fungal, presticide residue

10

15

20

25

30

35ร้อ

ยละท

ีไม่ผ่าน

เกณฑ์

0

5

สารเร่งเนือแดง

บอแรกซ์ สารฟอกขาว

สารกนัรา ฟอร์มาลีน สารเคมีป้องกนักาํจดั

ศตัรูพืช

นํามนัทอดซํา

Coliform bacteria ในนําบริโภค

Coliform bacteria ใน

นําแขง็

อาหารพร้อมบริโภค

มือผูส้มัผสัอาหาร

ภาชนะ อืน ๆ

ภาคกลาง 3.94 1.47 0.12 0.26 0.69 0.93 2.98 5.85 8.4 3.24 3.65 3.37 11.17ภาคเหนือ 0.06 0.03 0.03 0.67 1.16 1.64 9.42 1.06 3.56 6.39 5.91 6.59 2.88ภาคตะวนัออกเฉียงเหนือ 0 0.1 0.01 0.13 0.57 2.9 7.76 22.51 34.6 9.12 12.5 15.48 18.01ภาคใต้ 2.01 0.22 0.27 0.69 0.36 7.79 4.67 14.75 1.02 11.67 14.64 13.16 8.1

Risk based monitoring is needed?

risk-based food safety inspection during incident event in...

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