14-Jul 16-Jul 18-Jul 20-Jul 22-Jul 25-Jul 28-Jul

DAY 0 2 4 6 8 11 14
PARAOXON 23.5º PTE 196.46 193.67 189.98 194.42 203.71 194.73 194.39
198.15 191.91 192.91 198.29 203.00 194.91 192.80
Average 197.31 192.79 191.45 196.36 203.36 194.82 193.60
23.5º Control 229.59 230.00 229.20 225.45 228.77 227.95 228.41

4º PTE 198.04 175.09 186.80 205.38 198.19 202.14 190.69

198.96 192.66 194.76 195.18 204.59 197.88 200.13
Average 198.50 183.88 190.78 200.28 201.39 200.01 195.41
4º Control 230.22 230.00 230.19 230.36 229.69 227.82 229.03

-15.5º PTE 191.25 190.45 173.62 202.96 200.00 201.86 199.79

202.84 187.42 191.05 197.84 198.98 200.98 197.07
Average 197.05 188.94 182.34 200.40 199.49 201.42 198.43
-15.5º Control 228.48 231.03 232.38 226.98 229.84 229.47 230.69

PARATHION 23.5º PTE 204.50 190.67 193.72 183.20 202.10 196.80 204.51
197.23 195.85 193.84 202.23 208.75 198.57 201.43
Average 200.87 193.26 193.78 192.72 205.43 197.69 202.97
23.5º Control 233.95 231.30 229.17 225.33 227.47 227.65 227.02

4º PTE 197.78 196.38 195.10 205.60 202.69 204.44 205.70

202.40 194.57 192.47 197.51 196.67 199.43 201.30
Average 200.09 195.48 193.79 201.56 199.68 201.94 203.50
4º Control 229.64 231.81 229.55 231.86 231.23 230.50 230.60

-15.5º PTE 204.30 194.18 181.98 203.55 197.90 209.55 186.71

197.63 185.69 191.01 199.48 186.68 206.12 204.59
Average 200.97 189.94 186.50 201.52 192.29 207.84 195.65
-15.5º Control 229.66 231.67 230.59 227.90 230.72 229.32 228.10

0 2
COUMAPHOS 23.5º PTE 130.62 203.56 252.06 222.82 254.77 251.91 150.71
152.88 197.43 244.46 111.22 254.86 246.85 177.55
Average 141.75 200.50 248.26 167.02 254.82 249.38 164.13
23.5º Control 166.74 160.08 145.07 212.30 225.39 229.82 218.34

4º PTE 130.62 203.56 252.06 232.04 254.08 252.97 209.48

137.25 205.09 253.00 197.14 249.80 250.31 186.94
Average 133.94 204.33 252.53 214.59 251.94 251.64 198.21
4º Control 166.74 160.08 145.07 230.57 238.17 239.99 228.37

-15.5º PTE 143.18 205.18 252.04 218.62 254.27 251.47 185.17

152.88 197.43 244.46 208.59 254.90 251.68 208.97
Average 148.03 201.31 248.25 213.61 254.59 251.58 197.07
-15.5º Control 166.74 160.08 145.07 206.03 213.08 213.48 211.94
Notes:
- Using PTE purified from Pseudomonas putida, with pVLT-31 + opd gene
- Day 0 colour tested after 15 min incubation at approx. 5:30pm, thereafter colour allowed to develop over 15 min. Day 2 and
- Paraoxon and parathion blue light value in ImageJ used to represent change in yellow colour
- Coumaphos red, green, blue light colour values recorded to detect blue fluoresence change
- For coumaphos the photos taken typically show up a bright white blue fluorescence and in Image J the green and blue value

DAY 0 2 4 6 8 11 14
Coumaphos 23.5º PTE 1 1 1 1 1 1 1
(qualitative) 23.5º control 0 0 0 0 0 0 0
4º PTE 1 1 1 1 1 1 1
4 controlº 0 0 0 0 0 0 0
-15.5º PTE 1 1 1 1 1 1 1
-15.5º control 0 0 0 0 0 0 0
31-Jul 4-Aug 9-Aug 13-Aug 18-Aug 23-Aug
17 21 26 30 35 40
196.10 197.93 193.85 196.72 196.41 188.04
192.66 188.64 199.75 196.09 193.03 188.35
194.38 193.29 196.80 196.41 194.72 188.20
224.54 222.79 226.70 225.05 224.32 226.21

190.32 203.75 188.61 180.75 198.13 184.84

189.32 202.54 191.44 199.48 188.57 187.27
189.82 203.15 190.03 190.12 193.35 186.06
226.92 220.26 224.17 224.68 221.89 218.04

196.33 187.53 192.14 195.60 199.52 174.81

191.97 202.43 196.90 196.72 198.69 187.26
194.15 194.98 194.52 196.16 199.11 181.04
229.15 223.69 226.84 224.75 228.58 221.77

208.39 199.36 199.67 190.40 180.85 191.69

202.02 197.43 197.01 197.73 195.90 -
205.21 198.40 198.34 194.07 188.38 191.69
228.31 224.65 226.00 225.48 222.73 231.37

205.31 204.14 200.40 204.23 201.69 180.73

189.30 201.16 201.92 205.20 203.97 191.54
197.31 202.65 201.16 204.72 202.83 186.14
227.72 224.72 220.64 229.29 226.21 225.23

193.31 201.16 189.20 204.19 193.73 173.90

203.22 197.06 196.14 204.75 202.51 -
198.27 199.11 192.67 204.47 198.12 173.90
229.05 227.56 222.97 228.45 226.61 223.53

4 6 8
252.78 248.77 207.15 245.17 254.74 155.55 254.42 243.50 148.85 253.97
247.99 249.44 145.11 247.35 254.47 94.96 254.48 236.96 186.67 252.62
250.39 249.11 176.13 246.26 254.61 125.26 254.45 240.23 167.76 253.30
228.68 236.04 216.69 204.58 205.06 225.81 225.02 244.16 191.31 195.93

253.28 251.39 197.97 252.98 254.66 240.33 253.99 254.22 178.89 254.72
254.77 251.05 207.34 254.90 254.41 210.49 254.34 254.47 188.35 254.58
254.03 251.22 202.66 253.94 254.54 225.41 254.17 254.35 183.62 254.65
234.84 238.89 224.65 215.30 219.35 230.40 227.23 241.81 201.12 242.69

253.14 250.38 207.94 254.94 254.77 215.08 254.02 254.25 109.32 254.92
252.47 251.79 156.90 253.70 254.85 148.41 252.83 254.21 217.47 254.86
252.81 251.09 182.42 254.32 254.81 181.75 253.43 254.23 163.40 254.89
225.18 235.46 197.54 187.67 192.50 205.44 196.99 210.73 130.62 153.37
p over 15 min. Day 2 and 4 were incubated longer, approx 30 min…

J the green and blue values max out at 255.

Preformed by Chris Langley and David Collings

17 21 26 30 35 40
1 1 1 1 1 1
0 0 0 0 0 0
1 1 1 1 1 1
0 0 0 0 0 0
1 1 1 1 1 1
0 0 0 0 0 0
 11 14 17 21
242.83 244.70 253.30 252.20 215.94 253.24 250.38 179.84 253.95 250.95
246.15 240.40 252.60 252.30 180.03 252.00 246.00 159.69 251.54 248.51
244.49 242.55 252.95 252.25 197.99 252.62 248.19 169.77 252.75 249.73
192.73 246.20 241.60 221.60 232.08 230.37 214.20 192.37 190.63 178.24

245.80 227.50 247.10 248.00 192.80 249.90 248.50 190.50 254.03 250.60
247.10 233.80 252.80 250.50 217.85 254.60 250.00 211.54 253.77 251.22
246.45 230.65 249.95 249.25 205.33 252.25 249.25 201.02 253.90 250.91
247.80 243.42 237.45 216.54 222.17 219.83 205.43 197.46 198.29 192.50

238.38 187.80 253.90 247.00 213.20 254.33 250.27 161.26 254.91 249.73
249.85 214.44 252.03 249.65 194.17 251.84 247.95 173.71 254.63 250.53
244.12 201.12 252.97 248.33 203.69 253.09 249.11 167.49 254.77 250.13
174.17 236.50 230.90 213.80 220.32 215.83 203.32 186.81 186.52 179.16
Langley and David Collings beginning July 14th, 2010
 26 30 40
162.41 254.10 248.86 205.10 254.00 250.00
163.99 248.00 250.00 200.00 254.00 250.00
163.20 251.05 249.43 202.55 254.00 250.00
194.32 185.05 167.38 192.20 190.20 178.90

192.08 254.80 250.00 191.00 249.00 249.00

229.30 254.80 252.10 180.50 249.70 249.60
210.69 254.80 251.05 185.75 249.35 249.30
212.52 205.00 193.50 223.00 217.50 199.00
.
193.90 251.70 250.10 222.76 254.60 251.80
208.20 254.40 250.83 221.00 254.00 251.00
201.05 253.05 250.47 221.88 254.30 251.40
229.30 221.70 198.70 219.50 214.60 198.00
 Immobilized PTE Stability Test - 1mM Paraoxon

170.00
23.5º PTE
4º PTE
180.00 -15.5º PTE
23.5º Control
4º Control
190.00
-15.5º Control
Mean blue histogram value

200.00

210.00

220.00

230.00

240.00
0 5 10 15 20 25 30 35 40

Days
 Immobilized PTE Stability Test - 1mM Parathion

170.00

23.5º PTE

180.00 4º PTE
-15.5º PTE
23.5º Control
190.00 4º Control
Mean blue histogram value

-15.5º Control

200.00

210.00

220.00

230.00

240.00
0 5 10 15 20 25 30 35 40

Days
 1mM COUMAPHOS Blue

0 5 10 15 20 25 30 35
0.00

-15.5º Control
23.5 PTE
50.00
4 PTE
-15.5 PTE
4 Control
100.00
Mean blue detection

23.5 Control

150.00

200.00

250.00

300.00

Days
 Immobilized PTE Stability Test - 1mM Coumaphos (Qualitative)
1.2

1
Visible fluorescence under UV handheld

0.8

23.5º PTE
0.6 4º PTE
-15.5º PTE
23.5º control
0.4 4 controlº
-15.5º control

0.2

-1.66533453693773E-16
0 5 10 15 20 25 30 35 40

-0.2

Days