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Chapter 2 Table 2.1: Strain characteristics of all 13 strains. Strain Characteristics Strain Serovar Antigenic Structure Phage type PFGE Profile† Origin 27853 P. aeruginosa n/a n/a n/a ATCC Control Strain S08-0601 S. Agona 4,12:f,g,s PT39 1 SAGOXB.0066 3 Human faecal sample S09-0494 S. Agona 4,12:f,g,s SAGOXB.0066 3 Environmental swab from food plant S09-0046 S. Agona 4,12:f,g,s 1 Band variant to SAGOXB.0066 Drain swab S09-0371 S. Agona 4,12:f,g,s IE09019-14 4 Human faecal sample S09-0479 S. Agona 4,12:f,g,s PT3 1 IE09021-07 5 Sesame seed sample SL483 S. Agona 4,12,f,g,s 25.H.03 6 Control strain Accession No:CP001138 S09-0419 S. Typhimurium 4,5,12:i:- DT193 2 STYMXB.0131 7 Porcine carcass swab S08-0408 S. Typhimurium 4,5,12:i:2 DT193 2 STYMXB.0131 7 Human faecal sample SL1344 S. Typhimurium n/a n/a Control strain Accession No:FQ312003 PB449/LT2 S. Typhimurium n/a n/a Control Strain LT2 S09-0717 S. Enteritidis 9,12:g,m PT14b 1 n/a Human faecal sample S09-0004 S. Enteritidis 9,12:g,m PT4 1 n/a Imported Brazilian chicken Table 2.1 describes the characteristics of all 13 strains examined in this thesis, the strain collection incorporated 1 Pseudomonas aeruginosa, 6 Salmonella enterica subspecies enterica serovar Agona, 4 S. Typhimurium and 2 S. Enteritidis strains. 1 Denotes the phage type (PT) of the strain. 2 Denotes the definitive phage type (DT) of the strain. 3 Denotes the profile of the strain based on the Pulse Field Gel Electrophoresis (PFGE) assessment. 4-5 Denotes the PFGE reference number for the S. Agona strains not related to the SAGOXB.0066 strains. 6 PFGE profile produced in silico by Mark Achtman Group [185]. 7 Denotes the PFGE profile of the two S. Typhimurium strains examined in this thesis. See Appendix 3 for PFGE images supplied by the NSSLRL. Page 55
Chapter 2 2.5. Methods 2.5.1. 48 hour biofilm development method I. All experiments were performed in accordance with the current good laboratory practices and health and safety requirements outlined by National University of Ireland, Galway. II. III. IV. The test organisms were stored at -20°C on glycerol protect beads between experiments. Before each experiment the test strain was defrosted on the bench and streaked on Tryptic Soy Agar (TSA) and Xylose Lysine Deoxychocolate (XLD) plates which were incubated for 24 hours at 37°C.Isolated colonies were picked off the TSA plate and made to a 0.5 McFarland suspension (~1.5x10 8 CFU/ml) in sterile saline. 1ml of the suspension was added to 9ml of sterile Tryptic Soy Broth (TSB) and incubated in shaking incubator for 24 hours at 37°C. V. Twenty coupons (four of each of the five coupon types - glass, steel, polycarbonate, tile and concrete) were cleaned for use in each reactor run. VI. Coupons were sonicated at room temperature for 30 minutes in a detergent solution (2% Micro-90). VII. Coupons were then rinsed and sonicated for an additional 30 minutes in reagent-grade water. VIII. Using a 24-well plate the coupons were immersed in 1ml of 100% ethanol for 2 hours, ethanol was chosen as opposed to 2M HCl as this may chemical may result in degradation of surfaces i.e. rusting of stainless steel. IX. The coupons were visually inspected to ensure there was no degradation prior to use. X. The reactor was cleaned with micro-90 and ethanol prior to use. XI. The coupons were fixed into the reactor rods using a sterile forceps and set-screw, with the test area of the coupon (face) was placed inward exposed to the sheer force provided by the baffler rotator (see Figure 2.1). Page 56
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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
Page 26 and 27: Chapter 1 cracks or penetration of
Page 28 and 29: Chapter 1 detected by agglutination
Page 30 and 31: Chapter 1 become colonized with >10
Page 32 and 33: Chapter 1 antigens [z27],[z45] [1].
Page 34 and 35: Chapter 1 S. Agona has also been im
Page 36 and 37: Chapter 1 isolated from undercooked
Page 38 and 39: Chapter 1 The S. Typhimurium strain
Page 40 and 41: Chapter 1 ecosystem with the use of
Page 42 and 43: Chapter 1 [82]. Previous research h
Page 44 and 45: Chapter 1 to genes involved in flag
Page 46 and 47: Chapter 1 observation has also been
Page 48 and 49: Chapter 1 [119] and flow cell react
Page 50 and 51: Chapter 1 based disinfectant) the s
Page 52 and 53: Chapter 1 formation of two strains
Page 54 and 55: Chapter 1 shield the S. Agona cells
Page 56 and 57: Chapter 1 Key Objectives of this st
Page 58 and 59: Chapter 2 2. Abstract Food-borne pa
Page 60 and 61: Chapter 2 indicated that there may
Page 62 and 63: Chapter 2 taken from industrial set
Page 64 and 65: Chapter 2 2.1.4. CDC Biofilm Reacto
Page 66 and 67: Chapter 2 can be performed without
Page 68 and 69: Chapter 2 To summarise, previous au
Page 70 and 71: Chapter 2 2.3. Methods for examinin
Page 72 and 73: Chapter 2 and the biofilm formed th
Page 74 and 75: Chapter 2 2.4. Statistical Analysis
Page 78 and 79: Chapter 2 XII. The gas port and med
Page 80 and 81: Chapter 2 II. Primary fixative cons
Page 82 and 83: Chapter 2 IX. Once the conditions w
Page 84 and 85: Chapter 2 Figure 2.1: Image of CBR
Page 86 and 87: Chapter 2 2.6. Results 2.6.1. Asses
Page 88 and 89: Chapter 2 2.6.3. SEM Analysis of su
Page 90 and 91: Chapter 2 Figure 2.6: Complete remo
Page 92 and 93: Chapter 2 2.6.5. Mean log 10 densit
Page 94 and 95: Chapter 2 2.6.6. Mean log 10 densit
Page 96 and 97: Chapter 2 Table 2.5: Mean log 10 de
Page 98 and 99: Chapter 2 The results displayed in
Page 100 and 101: Chapter 2 Table 2.7: Difference bet
Page 102 and 103: Chapter 2 Table 2.8: The mean log 1
Page 104 and 105: Chapter 2 Figure 2.7: Graph of the
Page 106 and 107: Chapter 2 also used “high” soni
Page 108 and 109: Chapter 2 2.8. Summary All 13 strai
Page 110 and 111: Chapter 3 3. Abstract It has been e
Page 112 and 113: Chapter 3 classified as persistent
Page 114 and 115: Chapter 3 was plated onto TSA for e
Page 116 and 117: Chapter 3 Figure 3.1: The mean log
Page 118 and 119: Chapter 3 Figure 3.3: SEM image of
Page 120 and 121: Chapter 3 Table 3.2: Difference bet
Page 122 and 123: Chapter 3 3.4.3. Intra-serovar vari
Page 124 and 125: 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,
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Chapter 4 Dey/Engley (Difco) neutra
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Chapter 4 sterilisation was achieve
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Chapter 4 IX. Aliquots of 100µl of
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Chapter 4 XV. XVI. XVII. XVIII. XIX
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Chapter 4 4.3. Results 4.3.1. Suspe
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Chapter 4 Table 4.3: Mean log 10 de
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Chapter 4 Benzalkonium chloride was
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Chapter 4 4.4. Discussion Suspensio
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Chapter 4 effective at eliminating
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Chapter 4 Surprisingly, Nguyen et a
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Chapter 4 though a contact time of
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Chapter 4 provide a more standardis
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Chapter 4 hours instead of the stan
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Chapter 5 5. Abstract Examining bio
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Chapter 5 dry and rough (bdar) morp
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Chapter 5 absence of curli and cell
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Chapter 5 studies such as the micro
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Chapter 5 discussed in chapter 4. T
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Chapter 5 5.2. Methods 5.2.1. Colon
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Chapter 5 XIII. XIV. XV. XVI. XVII.
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Chapter 5 XVI. XVII. XVIII. XIX. XX
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Chapter 5 5.3.2. Repeatability of m
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Chapter 5 Figure 5.2: Stained biofi
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Chapter 5 was formed at room temper
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Chapter 5 Table 5.3: The density of
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Chapter 5 5.3.4. The density of bio
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Chapter 5 Table 5.6: The difference
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Chapter 5 Table 5.7: The difference
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Chapter 5 Table 5.8: Summary of ass
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Chapter 5 interpretation therefore
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Chapter 5 measures of S. enterica b
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Chapter 5 cells peaks [165, 191]. D
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Chapter 5 5.5. Limitations of the s
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Chapter 6 Discussion of S. enterica
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Chapter 6 However, a number of auth
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Chapter 6 It is of interest to exam
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Chapter 6 were similar at 48 hours.
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Chapter 6 attributable to a small n
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Chapter 6 chosen as an appropriate
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Chapter 6 undetermined. However, th
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Chapter 6 reason for this may be du
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Chapter 6 Moreover, based on the re
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Chapter 6 responsible for increased
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Chapter 6 not fully capture the ran
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Chapter 6 However, the extent of bi
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Chapter 6 6.10. Conclusions and rec
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Chapter 6 studies may solve some of
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Bibliography Bibliography Page 221
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Bibliography 12. T. Takata, J.L., H
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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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