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Chapter 5 5.3.4. The density of biofilm formed over 168 hours using the microtitre plate system When S. enterica biofilm was developed over an extended period of time (168 hours at 22°C), a similar pattern emerged in that the OD 590 readings for replicate wells of the same strain indicated that there was a large variation in repeated measurements. As displayed in Table 5.5, the mean OD 590 and the standard deviation is underlined for 5 S. Agona strains due to a small number of high OD 590 values for some observations for these strains. The mean OD 590 of stained biofilm would be smaller if these outliers were excluded. The full set of observations for all 13 strains is provided in Appendix 2- Table 4. When the mean density of biofilm formed by the strains was examined after 168 hours incubation there was evidence to suggest that some strains of S. Agona may form a more dense biofilm but overall this pattern was not consistent for all strains after incubation at 22°C. This trend was also not consistent for some strains when incubated at 37°C for the 168 hour duration (Table 5.6) most likely due to high and low outliers in some of the repeated reading of the OD 590 of S. Agona biofilm. The full set of observations are provided in Appendix 2 – Table 5. If the strains were to be grouped into the respective serovars as displayed in Table 5.6, the S. Agona serovar has the largest mean OD 590 for all 3 serovars. As previously discussed, because of the variation in OD 590 readings within individual strains caution should be exercised when grouping the strains together to compare the three S. enterica serovars. Page 177
Chapter 5 Table 5.5: The Density of biofilm formed over 168 hours Strain Number Organism 168 hours- 22°C 168 hours -37°C N Rank Mean SD Rank Mean SD 27853 P. aeruginosa 9 3 0.329 0.08 3 0.788 0.55 S08-0601 S. Agona Outbreak 9 10 0.193 0.12 4 0.699 0.89 S09-0494 S. Agona Outbreak 9 5 0.302 0.36 1 1.200 1.32 S09-0046 S. Agona variant 9 6 0.280 0.25 6 0.334 0.79 S09-0371 S. Agona 9 2 0.360 0.70 9 0.234 0.21 S09-0479 S. Agona 9 4 0.308 0.28 11 0.146 0.11 SL483 S. Agona 9 8 0.233 0.17 2 1.162 1.34 S09-0419 S. Typhimurium 9 1 0.368 0.15 10 0.147 0.17 S08-0408 S. Typhimurium 9 7 0.251 0.17 13 0.095 0.06 SL1344 S. Typhimurium 9 12 0.173 0.08 12 0.099 0.06 LT2 S. Typhimurium 9 13 0.166 0.09 5 0.519 0.69 S09-0717 S. Enteritidis 9 9 0.199 0.13 7 0.294 0.38 S09-0004 S. Enteritidis 9 11 0.173 0.09 8 0.235 0.26 Table 5.5 displays the mean OD 590 of biofilm formed on the surface of a microtitre plate following incubation at room temperature and 37°C for 168 hours. Strains include 1 Pseudomonas aeruginosa, 6 Salmonella enterica subspecies enterica serovar Agona, 4 Salmonella Typhimurium and 2 Salmonella Enteritidis strains. Table 5.5 also displays the mean and standard deviation of the mean which was calculated on 9 observations OD 590 (3 plates x 3 replicate wells in each plate). The most dense biofilm former (based on mean) is ranked as number one followed the next strain to form a dense biofilm et cetera. The mean OD 590 and standard deviation is underlined for 6 strains due to the large OD 590 for some observations for these strains. The full set of observations for all OD 590 measurements for all 13 strains is provided in Appendix 2- Table 4 and Table 5. Page 178
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Salmonella enterica - biofilm forma
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Table of Contents Page Number 1.16.
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Table of Contents Page Number 4.1.2
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Table of Contents Page Number 6.8.4
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List of Abbreviations Table Page Nu
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List of Abbreviations List of Abbre
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Summary of Content Summary of Conte
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“To show your true ability is alw
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Acknowledgements ever met, Fiona th
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Acknowledgements This Ph.D. thesis
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Chapter 1 1.1. Salmonella Salmonell
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Chapter 1 may impose enormous costs
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Chapter 1 cracks or penetration of
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Chapter 1 detected by agglutination
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Chapter 1 become colonized with >10
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Chapter 1 antigens [z27],[z45] [1].
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Chapter 1 S. Agona has also been im
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Chapter 1 isolated from undercooked
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Chapter 1 The S. Typhimurium strain
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Chapter 1 ecosystem with the use of
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Chapter 1 [82]. Previous research h
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Chapter 1 to genes involved in flag
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Chapter 1 observation has also been
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Chapter 1 [119] and flow cell react
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Chapter 1 based disinfectant) the s
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Chapter 1 formation of two strains
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Chapter 1 shield the S. Agona cells
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Chapter 1 Key Objectives of this st
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Chapter 2 2. Abstract Food-borne pa
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Chapter 2 indicated that there may
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Chapter 2 taken from industrial set
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Chapter 2 2.1.4. CDC Biofilm Reacto
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Chapter 2 can be performed without
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Chapter 2 To summarise, previous au
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Chapter 2 2.3. Methods for examinin
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Chapter 2 and the biofilm formed th
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Chapter 2 2.4. Statistical Analysis
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Chapter 2 Table 2.1: Strain charact
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Chapter 2 XII. The gas port and med
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Chapter 2 II. Primary fixative cons
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Chapter 2 IX. Once the conditions w
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Chapter 2 Figure 2.1: Image of CBR
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Chapter 2 2.6. Results 2.6.1. Asses
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Chapter 2 2.6.3. SEM Analysis of su
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Chapter 2 Figure 2.6: Complete remo
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Chapter 2 2.6.5. Mean log 10 densit
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Chapter 2 2.6.6. Mean log 10 densit
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Chapter 2 Table 2.5: Mean log 10 de
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Chapter 2 The results displayed in
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Chapter 2 Table 2.7: Difference bet
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Chapter 2 Table 2.8: The mean log 1
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Chapter 2 Figure 2.7: Graph of the
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Chapter 2 also used “high” soni
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Chapter 2 2.8. Summary All 13 strai
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Chapter 3 3. Abstract It has been e
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Chapter 3 classified as persistent
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Chapter 3 was plated onto TSA for e
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Chapter 3 Figure 3.1: The mean log
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Chapter 3 Figure 3.3: SEM image of
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Chapter 3 Table 3.2: Difference bet
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Chapter 3 3.4.3. Intra-serovar vari
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Chapter 3 3.4.4. Strain variation i
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Chapter 3 3.4.5. The density of bio
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Chapter 3 3.5. Discussion As illust
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Chapter 3 As discussed in section 3
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Chapter 3 increased biofilm appeare
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Chapter 4 Examining the efficacy of
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Chapter 4 through laboratory based
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Chapter 4 acid and quaternary ammon
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Chapter 4 the surface the temperatu
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Chapter 4 availability of multiple
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Chapter 4 peracetic acid (100, 200,
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Chapter 4 As discussed previously,
Page 148 and 149: Chapter 4 Dey/Engley (Difco) neutra
Page 150 and 151: Chapter 4 sterilisation was achieve
Page 152 and 153: Chapter 4 IX. Aliquots of 100µl of
Page 154 and 155: Chapter 4 XV. XVI. XVII. XVIII. XIX
Page 156 and 157: Chapter 4 4.3. Results 4.3.1. Suspe
Page 158 and 159: Chapter 4 Table 4.3: Mean log 10 de
Page 160 and 161: Chapter 4 Benzalkonium chloride was
Page 162 and 163: Chapter 4 4.4. Discussion Suspensio
Page 164 and 165: Chapter 4 effective at eliminating
Page 166 and 167: Chapter 4 Surprisingly, Nguyen et a
Page 168 and 169: Chapter 4 though a contact time of
Page 170 and 171: Chapter 4 provide a more standardis
Page 172 and 173: Chapter 4 hours instead of the stan
Page 174 and 175: Chapter 5 5. Abstract Examining bio
Page 176 and 177: Chapter 5 dry and rough (bdar) morp
Page 178 and 179: Chapter 5 absence of curli and cell
Page 180 and 181: Chapter 5 studies such as the micro
Page 182 and 183: Chapter 5 discussed in chapter 4. T
Page 184 and 185: Chapter 5 5.2. Methods 5.2.1. Colon
Page 186 and 187: Chapter 5 XIII. XIV. XV. XVI. XVII.
Page 188 and 189: Chapter 5 XVI. XVII. XVIII. XIX. XX
Page 190 and 191: Chapter 5 5.3.2. Repeatability of m
Page 192 and 193: Chapter 5 Figure 5.2: Stained biofi
Page 194 and 195: Chapter 5 was formed at room temper
Page 196 and 197: Chapter 5 Table 5.3: The density of
Page 200 and 201: Chapter 5 Table 5.6: The difference
Page 202 and 203: Chapter 5 Table 5.7: The difference
Page 204 and 205: Chapter 5 Table 5.8: Summary of ass
Page 206 and 207: Chapter 5 interpretation therefore
Page 208 and 209: Chapter 5 measures of S. enterica b
Page 210 and 211: Chapter 5 cells peaks [165, 191]. D
Page 212 and 213: Chapter 5 5.5. Limitations of the s
Page 214 and 215: Chapter 6 Discussion of S. enterica
Page 216 and 217: Chapter 6 However, a number of auth
Page 218 and 219: Chapter 6 It is of interest to exam
Page 220 and 221: Chapter 6 were similar at 48 hours.
Page 222 and 223: Chapter 6 attributable to a small n
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Page 226 and 227: Chapter 6 undetermined. However, th
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Page 230 and 231: Chapter 6 Moreover, based on the re
Page 232 and 233: Chapter 6 responsible for increased
Page 234 and 235: Chapter 6 not fully capture the ran
Page 236 and 237: Chapter 6 However, the extent of bi
Page 238 and 239: Chapter 6 6.10. Conclusions and rec
Page 240 and 241: Chapter 6 studies may solve some of
Page 242 and 243: Bibliography Bibliography Page 221
Page 244 and 245: Bibliography 12. T. Takata, J.L., H
Page 246 and 247: Bibliography 29. Barker, J., M. Nae
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Bibliography 47. Threlfall, E.J., H
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Bibliography 65. CDC. Investigation
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Bibliography 83. Ledeboer, N., Frye
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Bibliography 100. Chia, T., Goulter
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Bibliography 117. Buckingham-Meyer,
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Bibliography 133. European Committe
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Bibliography Supernatant of a Hafni
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Bibliography 165. Díez-García, M.
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Bibliography cultivation of Staphyl
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Bibliography Compounds. 2012. Appli
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Bibliography 215. Buck, et al., A q
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Bibliography Canadian Journal of Ps
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Appendix 1 Appendix 1— List of Re
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Appendix 1 Appendix 1 - List of Equ
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Appendix 2 Colour index High outlie
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Appendix 3 Appendix 3-Figure 2: PFG
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Appendix 4 Poster Presentation Envi
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