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14-Jul

PARAOXON

DAY
23.5º PTE

Average

18-Jul

20-Jul

22-Jul

25-Jul

28-Jul

2

4

6

8

11

14

196.46
198.15

193.67
191.91

189.98
192.91

194.42
198.29

203.71
203.00

194.73
194.91

194.39
192.80

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
198.96

175.09
192.66

186.80
194.76

205.38
195.18

198.19
204.59

202.14
197.88

190.69
200.13

198.50

183.88

190.78

200.28

201.39

200.01

195.41

230.22

230.00

230.19

230.36

229.69

227.82

229.03

191.25
202.84

190.45
187.42

173.62
191.05

202.96
197.84

200.00
198.98

201.86
200.98

199.79
197.07

197.05

188.94

182.34

200.40

199.49

201.42

198.43

228.48

231.03

232.38

226.98

229.84

229.47

230.69

204.50
197.23

190.67
195.85

193.72
193.84

183.20
202.23

202.10
208.75

196.80
198.57

204.51
201.43

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
202.40

196.38
194.57

195.10
192.47

205.60
197.51

202.69
196.67

204.44
199.43

205.70
201.30

200.09

195.48

193.79

201.56

199.68

201.94

203.50

229.64

231.81

229.55

231.86

231.23

230.50

230.60

204.30
197.63

194.18
185.69

181.98
191.01

203.55
199.48

197.90
186.68

209.55
206.12

186.71
204.59

200.97

189.94

186.50

201.52

192.29

207.84

195.65

229.66

231.67

230.59

227.90

230.72

229.32

228.10

130.62
152.88

203.56
197.43

0
252.06
244.46

222.82
111.22

254.77
254.86

2
251.91
246.85

150.71
177.55

Average
4º Control
-15.5º PTE

Average
-15.5º Control
PARATHION

16-Jul
0

23.5º PTE

Average

Average
4º Control
-15.5º PTE

Average
-15.5º Control

COUMAPHOS 23.5º PTE

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
137.25

203.56
205.09

252.06
253.00

232.04
197.14

254.08
249.80

252.97
250.31

209.48
186.94

133.94

204.33

252.53

214.59

251.94

251.64

198.21

166.74

160.08

145.07

230.57

238.17

239.99

228.37

143.18
152.88

205.18
197.43

252.04
244.46

218.62
208.59

254.27
254.90

251.47
251.68

185.17
208.97

148.03

201.31

248.25

213.61

254.59

251.58

197.07

166.74

160.08

145.07

206.03

213.08

213.48

211.94

Average
4º Control
-15.5º PTE

Average
-15.5º Control

5º PTE 23.Day 0 colour tested after 15 min incubation at approx.Using PTE purified from Pseudomonas putida.5º control 4º PTE 4 controlº -15. blue light colour values recorded to detect blue fluoresence change .5º control 0 1 0 1 0 1 0 2 1 0 1 0 1 0 4 1 0 1 0 1 0 6 1 0 1 0 1 0 8 1 0 1 0 1 0 11 1 0 1 0 1 0 14 1 0 1 0 1 0 .For coumaphos the photos taken typically show up a bright white blue fluorescence and in Image J the green and blue value Coumaphos (qualitative) DAY 23.Paraoxon and parathion blue light value in ImageJ used to represent change in yellow colour . Day 2 and . 5:30pm. green. thereafter colour allowed to develop over 15 min.5º PTE -15. with pVLT-31 + opd gene .Notes: .Coumaphos red.

39 249.66 197.21 109.55 94.26 254.48 198.77 208.41 194.43 192.81 181.85 215.09 196.65 226.16 199.33 210.25 254.70 254.08 148.23 193.57 184.04 196.06 225.33 191.75 228.31 189.21 225.54 225.14 196.22 254.45 240.54 188.51 - 173.16 204.19 204.45 226.52 196.81 187.31 224.83 254.38 193.30 204.61 191.41 254.65 201.05 197.34 252.90 - 191.69 204.31-Jul 4-Aug 9-Aug 13-Aug 18-Aug 23-Aug 17 21 26 30 35 40 196.89 225.04 188.74 254.35 194.34 254.84 187.62 250.98 254.90 229.67 192.27 199.43 254.04 216.75 199.62 153.96 148.97 228.16 191.78 247.75 202.09 182.34 194.23 167.56 222.06 189.10 192.38 251.31 195.96 254.22 202.54 187.62 254.42 254.69 228.23 163.44 180.72 224.27 189.05 227.75 253.99 4 248.69 253.31 202.89 218.02 244.65 215.85 195.65 234.44 196.11 176.61 125.73 191.40 197.72 202.01 190.79 226.69 226.41 240.42 254.15 194.81 225.73 231.80 196.12 242.32 203.53 252.41 254.37 .16 197.32 254.12 173.11 192.20 224.13 246.36 197.48 222.39 202.03 188.21 190.69 203.92 204.86 252.92 220.67 204.15 145.00 225.75 196.31 203.04 229.85 186.26 254.30 228.67 197.14 201.66 254.98 194.18 235.79 207.60 196.84 238.47 250.82 203.05 224.29 196.07 188.61 223.11 245.50 205.54 197.44 207.23 205.92 254.66 253.12 193.38 191.83 186.58 254.90 195.73 130.22 201.72 196.20 201.89 188.67 253.54 222.53 202.70 225.97 180.68 236.97 252.03 190.99 254.32 189.64 193.39 251.35 6 254.77 251.35 183.90 254.75 193.77 254.28 254.14 252.85 199.48 8 243.72 188.47 254.14 204.72 220.40 227.47 178.97 187.14 227.94 253.43 199.93 253.03 251.02 252.20 196.37 205.68 221.32 226.73 202.90 254.77 249.46 197.40 254.81 201.84 224.17 254.81 251.93 188.15 190.17 224.35 186.69 205.02 199.13 188.26 194.40 201.94 254.26 224.90 198.35 230.30 219.58 221.49 253.69 174.15 223.73 180.89 224.94 156.50 236.40 198.76 253.35 254.41 193.11 181.21 198.72 199.52 198.47 155.97 207.23 241.47 198.64 229.16 200.32 217.72 254.29 226.06 226.58 205.17 247.99 210.

Day 2 and 4 were incubated longer.p over 15 min. approx 30 min… J the green and blue values max out at 255. Preformed by Chris Langley and David Collings 17 1 0 1 0 1 0 21 1 0 1 0 1 0 26 1 0 1 0 1 0 30 1 0 1 0 1 0 35 1 0 1 0 1 0 40 1 0 1 0 1 0 .

85 249.00 249.80 243.95 249.45 230.13 174.63 249.80 247.65 249.45 216.50 230.25 249.90 252.51 244.00 250.00 17 250.63 178.08 230.90 254.27 247.11 242.20 192.03 253.50 238.50 192.73 250.83 203.30 215.20 241.43 197.99 252.09 249.11 167.81 186.25 201.80 217.46 198.55 252.54 222.20 194.12 201.38 246.49 242.53 244.80 247.65 213.15 244.80 220.50 233.70 240.60 248.83 246.91 247.24 245.33 251.77 250.25 205.50 250.95 248.25 197.33 252.77 252.83 205.32 215.71 254.12 252.03 253.49 254.54 254.52 179.24 252.60 221.10 252.80 248.00 179.10 227.29 192.22 246.40 253.44 253.37 190.75 249.95 161.95 251.73 246.02 253.69 253.60 14 252.42 237.60 251.16 .38 249.30 252.32 186.19 169.95 252.84 250.90 250.97 248.54 21 250.03 247.94 180.26 173.20 252.69 253.50 211.84 159.60 232.73 192.33 203.00 190.85 187.80 214.37 214.17 219.91 254.62 248.90 213.17 254.77 250.17 236.

Langley and David Collings beginning July 14th. 2010 .

30 251.00 194.70 249.05 249.43 202.00 250.00 251.00 249.00 205.00 193.00 252.00 .32 185.76 221.70 254.60 198.00 249.99 254.05 250.55 254.50 249.50 199.80 254.83 222.60 210.52 205.80 251.10 200.00 193.75 249.05 253.80 250.40 250.00 254.20 251.90 208.80 251.10 250.86 250.00 250.50 223.50 214.20 190.60 254.00 254.90 192.35 249.00 201. 40 .10 248.20 251.20 178.00 254.00 30 250.88 254.40 229.05 167.30 221.162.10 191.70 219.00 180.70 198.30 212.30 254.05 185.38 192.00 217.69 254.41 163.00 26 248.47 221.08 229.00 163.

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5º Control .00 Mean blue histogram value 180.5º Control 4º Control -15.00 Days 4º PTE -15.00 23.00 240.Immobilized PTE Stability Test .00 190.00 220.00 200.5º PTE 210.00 230.1mM Paraoxon 0 10 20 30 40 170.5º PTE 23.

5º PTE 210.00 220.00 200.1mM Parathion 0 10 20 30 40 170.00 23.00 190.00 230.5º Control .5º PTE 23.00 Days 4º PTE -15.00 Mean blue histogram value 180.00 240.5º Control 4º Control -15.Immobilized PTE Stability Test .

00 -15.00 250.1mM COUMAPHOS Blue 0 5 10 15 20 25 30 35 0.5º Control 150.00 300.00 200.00 Mean blue detection 50.00 100.00 Days Row 36 Row 41 Row 46 Row 42 Row 37 .

2 0 5 10 15 20 Days 25 30 35 40 .5º control 0.8 23.2 Visible fluorescence under UV handheld 1 0.1mM Coumaphos (Qualitative) 1.5º PTE 4º PTE -15.4 0.6 0.Immobilized PTE Stability Test .2 0 -0.5º control 4 controlº -15.5º PTE 23.