1 improving food safety by poultry carcass mapping thomas p. oscar, ph.d. usda, ars princess anne,...
TRANSCRIPT
1
Improving Food Safety by
Poultry Carcass Mapping
Thomas P. Oscar, Ph.D.
USDA, ARS
Princess Anne, MD
2
3
Definition: Poultry Carcass Map
• Map of the distribution of pathogens on the carcass of poultry.
Salmonella Campylobacter Listeria
4
• Location
• Number
• Types
Military Map: War on Pathogens
5
Why Poultry Carcass Maps?
To better assess and manage food safety risks!
6
Microbial EcologyPathogen is a minority member of the microbial community
– Unattached
• surface water layer
– Attached
• biofilms
– Entrapped
• skin crevices
• feather follicles
• deep tissues
7
Carcass Sampling Methods
• Rinse
• Swab
• Sponge
• Excision
These methods will not produce an accurate carcass map
8
Whole Carcass Enrichment
• Detects all pathogens present.
• Can be used to determine pathogen numbers.
9
= pathogen= competitor
= pre-enrichment broth= carcass part
< 1 CFU/ml > 3 CFU/ml
Whole Carcass Enrichment Concepts and principles for enumeration
10
Whole Sample EnrichmentStandard Curve for Enumeration
0 1 2 3 4 5 6 70
5
10
15
20
25
Y = 1 + 4.89X - 0.31X2
R2 = 0.9611
Salmonella spp. (log number/25 g)
PC
R d
etec
tion
time
scor
e
Oscar, T. P. (2004) J. Food Prot. 67(6):1201-1208.
11
Poultry Carcass MappingConcepts and Principles
Cornish Game Hen
12
A: Wing, left41 g
B: Wing, right41 g
G: Back, rib67 g
H: Back, sacral77 g
C: Breast, left front62 g
D: Breast, right front60 g
E: Breast, left back35 g
F: Breast, right back37 g
I: Thigh, left48 g
J: Thigh, right50 g
K: Drumstick, left45 g
L: Drumstick, right48 g
A B
G
H
D
F
J
L
C
E
I
K
Step 1Divide Carcass into 12 Parts
13
A: Wing, left0.5-1 log
B: Wing, right0.5-1 log
G: Back, rib3.5-4 log
H: Back, sacral3.5-4 log
C: Breast, left front1.5-2 log
D: Breast, right front1.5-2 log
E: Breast, left back2.5-3 log
F: Breast, right back2.5-3 log
I: Thigh, left4.5-5 log
J: Thigh, right4.5-5 log
K: Drumstick, left5.5-6 log
L: Drumstick, right5.5-6 log
A B
G
H
D
F
J
L
C
E
I
K
Step 2Spot Inoculate with 1 to 106 CFU of a MDR Salmonella
2 μl 5 μl
14
Distribution of Salmonellain a 5 μl drop of a 3 log/ml diluted culture
0 1 2 3 4 5 6 7 8 9 10 11 12 13 140.00
0.05
0.10
0.15
0.20
No (CFU)
Fre
qu
ency
Oscar, T. P. (2009) J. Food Prot. 72:304-314
15
Step 3Incubate whole parts in 300 ml BPW at 40C and 100 rpm for 24 h
16
Step 4Sample and drop plate (2 μl) onto XLH-CATS @ 1, 2, 3, 4, 5, 6, 7, 8, 24
h
17
Step 5Incubate XLH-CATS plates @ 38C for 24 h
18
Step 6Capture the Image
19
Step 7Convert to Monochrome Image
A) 10-7 (3 log10 CFU/ml) B) 10-6 (4 log10 CFU/ml)
C) 10-5 (5 log10 CFU/ml)D) 10-4 (6 log10 CFU/ml)
20
A) 1 h B) 2 h C) 3 h
E) 5 h F) 6 hD) 4 h
G) 7 h H) 8 h I) 24 h
21
Step 8Count pixels
22
Rep4
0 4 8 12 16 20 240
100
200
300
400
500
600
700
a
b
c
d
e
f
g
h
I
jk
l
Time (h)
s165
(pi
xels
)
Step 9Determine Detection Time
23
0 1 2 3 4 5 6 7 8 9 100
1
2
3
4
5
6
7
8
9
10
DT (h)
No
(log
10)
No = 7.78 ± 0.61 – (0.995*DT)
Step 10Develop a Standard Curve
24
Limitation: Only MDR strainsSolution: Real-time PCR or Impedance
25
Qualitative MapCornish Game Hen @ Retail
26
A B
G
H
D
F
J
L
C
E
I
K
Qualitative Map
Obtain chickens from retail
Divide carcass into 12 parts
Incubate part in BPW (300 ml)
Transfer 0.1 ml of BPW enrichment to RV broth
Streak RV broth enrichment onto XLT4
Grow suspect colony in BHI broth
Store isolate at –70C
Serotype, R-type and pulsotype
27
Salmonella serotypeAntibiotic Resistance
Profile Thompson Kentucky Typhimurium Glostrup Enteritidis n %
Pansusceptable 1 1 3 5 2.8%
Ax-Am-Ce-Su-Te 1 136 137 75.7%
Ax-Am-Ce-K-Su-Te 24 24 13.3%
Am-Cm-Su-Te 7 7 3.9%
Ax-Am-Ce-Te 1 1 0.6%
Ax-Am-Ce-Cef-Su-Te 1 1 0.6%
G-Su-Te 4 4 2.2%
Ax-Am-Su-Te 1 1 0.6%
Ax-Am-Ce-Cef-K-Su-Te 1 1 0.6%
n 1 2 171 4 3 181
% 0.6% 1.1% 94.5% 2.2% 1.7%
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4/1
0/0
6
4/2
5/0
6
5/3
0/0
6
7/1
7/0
6
8/1
4/0
6
9/2
5/0
6
11
/13
/06
12
/4/0
6
1/2
2/0
7
2/2
0/0
7
3/2
6/0
7
4/2
3/0
7
2/1
4/2
00
8
3/1
1/2
00
8
4/2
0/2
00
8
5/2
0/2
00
8
6/2
9/2
00
8
9/1
4/2
00
8
0
5
10
15
Date
Co
nta
min
ated
Par
ts p
er C
arca
ss
29
0 1 2 3 4 5 6 7 8 9 10 11 120
10
20
30
40
Contaminated Parts per Carcass
Fre
qu
ency
Parts = 181/840 (21.5%)
Carcasses = 40/70 (57.1%)
37 different patterns
30
A B
G
H
D
F
J
L
C
E
I
K
April 17, 2006
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
April 25, 2006 May 22, 2006
A B
G
H
D
F
J
L
C
E
I
K
May 30, 2006
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
June 5, 2006 June 12, 2006
Pos.
Neg.
31
G: Back, ribThompson
Pansusceptable
A B
G
H
D
F
J
L
C
E
I
K
April 17, 2006
L: Drumstick, rightKentucky
AxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
C: Breast, left frontTyphimuriumAxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
April 25, 2006 May 22, 2006 Pos.
Neg.
32
A B
G
H
D
F
J
L
C
E
I
K
July 31, 2006
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
June 19, 2006
A B
G
H
D
F
J
L
C
E
I
K
Sept. 18, 2006
A B
G
H
D
F
J
L
C
E
I
K
July 17, 2006
A B
G
H
D
F
J
L
C
E
I
K
Sept. 25, 2006 Oct. 16, 2006
Pos.
Neg.
33
A: Wing, leftTyphimuriumPansusceptable
G: Back, ribTyphimurium
AxAmCeKSuTe
C: Breast, left frontKentucky
AxAmCeTe
A B
G
H
D
F
J
L
C
E
I
K
October 16, 2006
Pos.
Neg.
n =33 with > 1 contaminated part
Serotype 12.1% (4/33)
Ab resistance pattern 33.3% (11/33)
PFGE pattern 100% (33/33)
34
A B
G
H
D
F
J
L
C
E
I
K
Nov. 6, 2006
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
Nov. 13, 2006 Nov. 27, 2006
A B
G
H
D
F
J
L
C
E
I
K
Dec. 4, 2006
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
Dec. 4, 2006 Jan. 8, 2006
Pos.
Neg.
35
A: Wing, leftTyphimuriumAxAmCeSuTe
B: Wing, rightTyphimuriumAxAmCeSuTe
G: Back, ribTyphimuriumAxAmCeSuTe
H: Back, sacralTyphimuriumAxAmCeSuTe
L: Drumstick, rightTyphimuriumAxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
Dec. 4, 2006
PFGE patterns = 100% similarity
Pos.
Neg.
36
A B
G
H
D
F
J
L
C
E
I
K
Jan. 22, 2007
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
Feb. 5, 2007 Feb. 20, 2007
A B
G
H
D
F
J
L
C
E
I
K
Feb. 26, 2007
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
March 5, 2007 March 12, 2007
Pos.
Neg.
37
A: Wing, leftTyphimuriumAxAmCeSuTe
B: Wing, rightTyphimuriumAxAmCeSuTe
G: Back, ribTyphimuriumAxAmCeSuTe
H: Back, sacralTyphimuriumAxAmCeSuTe
C: Breast, left frontTyphimuriumAxAmCeSuTe
D: Breast, right frontTyphimuriumAxAmCeSuTe
E: Breast, left backTyphimuriumAxAmCeSuTe
F: Breast, right backTyphimuriumAxAmCeSuTe
K: Drumstick, leftTyphimuriumAxAmCeSuTe
L: Drumstick, rightTyphimuriumAxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
March 5, 2007Pos.
Neg.
PFGE patterns = 100% similarity
38
A B
G
H
D
F
J
L
C
E
I
K
March 26, 2007
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
April 2, 2007 April 9, 2007
A B
G
H
D
F
J
L
C
E
I
K
April 16, 2007
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
April 23, 2007 April 30, 2007
Pos.
Neg.
39
B: Wing, rightGlostrupGSuTe
G: Back, ribGlostrupGSuTe
C: Breast, left frontGlostrupGSuTe
D: Breast, right frontGlostrupGSuTe
L: Drumstick, rightTyphimuriumAxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
April 9, 2007Pos.
Neg.
40
A B
G
H
D
F
J
L
C
E
I
K
Jan. 28, 2008
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
Feb. 14, 2008 Feb. 28, 2008
A B
G
H
D
F
J
L
C
E
I
K
March 5, 2008
A B
G
H
D
F
J
L
C
E
I
K
A B
G
H
D
F
J
L
C
E
I
K
April 2, 2008 April 9, 2008
Pos.
Neg.
41
H: Back, sacralTyphimuriumAxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
April 2, 2008Pos.
Neg.
A: Wing, leftTyphimurium
AxAmCeCefKSuTe
H: Back, sacralTyphimuriumAxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
April 9, 2008
A: Wing, leftTyphimuriumAxAmCeSuTe
42
A B
G
H
D
F
J
L
C
E
I
K
April 20, 2008
A B
G
H
D
F
J
L
C
E
I
K
June 3, 2008
A B
G
H
D
F
J
L
C
E
I
K
July 14, 2008
A B
G
H
D
F
J
L
C
E
I
K
Sept. 20, 2008
Pos.
Neg.
43
H: Back, sacralTyphimuriumAxAmCeSuTe
D: Breast, right frontTyphimuriumAxAmCeSuTe
A B
G
H
D
F
J
L
C
E
I
K
April 20, 2008Pos.
Neg.
I: Thigh, leftEnteritidis
Pansusceptable
44
A: Wing, left41 g
27.1%
B: Wing, right41 g
24.3%
G: Back, rib67 g
38.6%
H: Back, sacral77 g
34.3%
C: Breast, left front62 g
28.6%
D: Breast, right front60 g
25.7%
E: Breast, left back35 g7.1%
F: Breast, right back37 g8.6%
I: Thigh, left48 g
17.1%
J: Thigh, right50 g
12.9%
K: Drumstick, left45 g
10.0%
L: Drumstick, right48 g
24.3%
A B
G
H
D
F
J
L
C
E
I
K
Population Map (n = 70) Poultry InspectionWPE of all edible parts
45
Wings versus Thighs
Fisher's exact test
P value 0.0371
P value summary *
One- or two-sided Two-sided
Significant? (P<0.05) Yes
Data analyzed Positive Neg. Total
A + B 36 104 140
I + J 21 119 140
Total 57 223 280
ExplanationCarcass hung upside down and washed from back to front
46
Left versus Right Drumstick
Fisher's exact test
P value 0.0420
P value summary *
One- or two-sided Two-sided
Significant? (P<0.05) Yes
Data analyzed Positive Neg. Total
K 7 63 70
L 17 53 70
Total 24 116 140
ExplanationViscera hung on right drumstick
47
Part code Part Weight, g Positive Total Incidence
Gskin Back, rib 5.2 0 6 0.0%
Ginside 62.9 5 6 83.3%
Fine Map: Sub-parts
ExplanationContamination from crop
48
Future Mapping Research
• Broilers and Turkeys
• Other Plants
• Process Steps
• Sub-part maps
• Other Pathogens
• Other Foods
• Quantitative
49
• Location
• Number
• Types
Take Home Message:Correct Approach = Accurate Map = Good Food Safety
Decisions = Improved Food Safety