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ability of Acinetobacter species to survive in human and environmental reservoirs in which the genes of resistance may be transferred (Vila 1998). For multi-drug resistant Acinetobacter infections, several studies have demonstrated clinical effectiveness of sulbactam in combination with ampicillin or cefoperazone. The only effective antibacterial agent to this bacterium is colistin (Winn et al., 2006). One report has demonstrated the efficiency and safety of colistin in patients with Acinetobacter infection that was not susceptible to carbapenem and shown that 57% of patients cured with colistin therapy, without prolonged neuro-muscular blockade as a side effect of therapy (Torres et al., 2007) Recently Enoch and his colleges (2008) completed a six month study of the outbreak caused by multi-drug-carbapenem-resistant Acinetobacter baumanii (MRAB-C) strain in UK hospitals. This outbreak would have increased the rate of mortality if not controlled properly. However, isolation of the patients infected with MRAB-C, education of staff dealing with those patients, early patient and environmental screening, and effective hygiene all helped to control this outbreak. Also, patients infected with MRAB-C were treated with colistin and tigecycline and improved. 1.3.8 Acinetobacter Treatment with Honey: Due to the emergence of bacteria resistant to antibiotics, the bactericidal properties of manuka honey have been extensively researched. Currently, few studies have reported the antibacterial activity of honey against Acinetobacter. George & Cutting, (2007) initiated an in-vitro study of the antibacterial activity of Medihoney against 130 clinical isolates of multi-drug resistant organisms including Acinetobacter. The study showed that the concentration needed to inhibit the resistant Acinetobacter strains was 8% (v/v) of honey. 50
More recently, a study was carried out using Malaysian tualang honey in comparison to manuka honey against wound pathogens including Acinetobacter. The antibacterial activity for both honeys was same with the MIC ranges between (11.25 & 12.5% w/v) (Tan et al., 2009). Although Acinetobacter spp. have been found to be susceptible to honey, more objective evidence derived from clinical trails and animal models to determine whether honey has a similar antimicrobial effect in-vivo are required. 1.4 Alternative Antimicrobial Therapies: As concerns about antimicrobial resistance increase, efforts of the pharmaceutical industry to develop new drugs have diminished and the possibility of running out of effective antimicrobial agents has increased. It takes 10-12 years for new antibiotic to be developed and costs approximately £250 million for each one (Greenwood 2003). Yet the chance of developing a new drug that will have excellent bactericidal properties without causing the emergence of resistance is impossible. The introduction and application of honey into clinical practice several years ago has put attention on using it for treatment of various infections. An extensive review of clinical studies by Molan suggested that honey might be successfully used as an antimicrobial agent and also in promoting healing of wounds (Molan 2006). 1.4.1 Ancient Use of Honey as a Medicine: Honey has been used for many thousands of years as a food, a medicine and it has been incorporated into cosmetics. A large number of different cultures have extensively used honey as a medicine for many disorders. It has been used in wound 51
Page 1 and 2: Honey as an antimicrobial agent aga
Page 3 and 4: 1.2.3 Emergence of ESBL: 9 1.2.4 Ri
Page 5 and 6: 2.2.1 Honey sample collection: 51 2
Page 7 and 8: 2.5 Effect of honey on bacterial st
Page 9 and 10: viable count 3.4.2.1 Acinetobacter
Page 11 and 12: Declaration This work has not previ
Page 13 and 14: Acknowledgments I wish to express m
Page 15 and 16: Index of Tables Tables Title Page T
Page 17 and 18: Index of Tables (Continued) Tables
Page 19 and 20: Index of Figures Figure Title Page
Page 21 and 22: Index of Figures (Continued) Figure
Page 27 and 28: ESBL- Extended spectrum beta lactam
Page 29 and 30: TEM- Temoneira UTI- Urinary tract i
Page 31 and 32: Chapter 1 Introduction 31
Page 33 and 34: such as methicillin resistant Staph
Page 35 and 36: Beta-lactamases in this classificat
Page 37 and 38: cefotaxime faster than ceftazidime;
Page 39 and 40: 1.2.3 Emergence of ESBL: The produc
Page 41 and 42: Acinetobacter spp are widespread in
Page 43 and 44: many secretions of normal people an
Page 45 and 46: 1.3.4.1 The production of exopolysa
Page 47 and 48: 1.3.5 Emergence of Resistance: A.ba
Page 49: In the last few years resistant str
Page 53 and 54: Gideonsen 2005), malignant ulcers (
Page 55 and 56: Nezeako & Hamdi, (2000) tested six
Page 57 and 58: In addition, honey contains several
Page 59 and 60: with the help of an enzyme (glucose
Page 61 and 62: (Wahdan 1998), and vanillic acid, c
Page 63 and 64: Table 1.2: Comparisons between pero
Page 65 and 66: Another study compared the effectiv
Page 67 and 68: of raw honey samples from Acacia, h
Page 69 and 70: 1.5 Aims and Objectives: The aim of
Page 71 and 72: The materials used throughout this
Page 73 and 74: Table 2.3: Chemicals and Reagents u
Page 75 and 76: Table 2.5: Equipment used in electr
Page 77 and 78: Fifty five clinical isolates belong
Page 79 and 80: 2.1 Characterization of test organi
Page 81 and 82: 2.2 Characterization of honey sampl
Page 83 and 84: transfered into a large square assa
Page 85 and 86: 2.2.3 Determination of pH: This met
Page 87 and 88: once the tubing opened at one end a
Page 89 and 90: was re-suspended in 1 ml liquefied
Page 91 and 92: difference between this absorbance
Page 93 and 94: and Omani honey (B) with hydrogen p
Page 95 and 96: 2.3.1.2 Broth dilution method: 2.3.
Page 97 and 98: 2.4 Time Kill Curve Assay: 2.4.1 St
Page 99 and 100: 2.5.1.2 Preparation of cells in the
Page 101 and 102:
2.5.2.2 Preparation of cells for tr
Page 103 and 104:
Table 2.14: Reagents and buffers us
Page 105 and 106:
samples (Fig 2.3). Each sample was
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tray was covered and wrapped in sar
Page 109 and 110:
Chapter 3 Results 109
Page 111 and 112:
Table 3.1a: Identification and anti
Page 113 and 114:
Table 3.2: Confirmation of identity
Page 115 and 116:
Page 117 and 118:
Page 119 and 120:
The diameter of the zones of inhibi
Page 121 and 122:
Table 3.7: Antibacterial activity o
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with visual estimation. The colour
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Figure 3.3: Image of pollen present
Page 127 and 128:
3.3 Determination of antibacterial
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3.3.1.2 Broth dilution method: The
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The MIC range of 30 Acinetobacter i
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Mean MIC and MBC (%w/v) values of m
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Table 3.15: Susceptibility of 15 En
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3.3.2 Sensitivity of MDR and ESBLs
Page 139 and 140:
Table 3.18: Susceptibility of 11 Kl
Page 141 and 142:
Table 3.20: Susceptibility of 15 En
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Table 3.22: Susceptibility of 8 Ser
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3.4 Time Kill Curves One representa
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Figure 3.6: The effect of manuka ho
Page 149 and 150:
3.4.2 Inhibition of test organisms
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This data was done in triplicate Nu
Page 153 and 154:
This data was done in triplicate Fi
Page 155 and 156:
Page 157 and 158:
Page 159 and 160:
Page 161 and 162:
Figure 3.15: The effect of manuka h
Page 163 and 164:
All isolates showed statistically s
Page 165 and 166:
Figure 3.16: Growth curve of Acinet
Page 167 and 168:
Figure 3.20: Growth curve of Citrob
Page 169 and 170:
(Table 3.26). The length of individ
Page 171 and 172:
Figure 3.24: SEM micrograph of untr
Page 173 and 174:
Figure 3.28: SEM micrographs of unt
Page 175 and 176:
C D Cell division Multiple septa in
Page 177 and 178:
Figure 3.31: SEM micrographs of Aci
Page 179 and 180:
Page 181 and 182:
3.5.2.3 SEM of Klebsiella Klebsiell
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Page 185 and 186:
Figure 3.39: SEM micrographs of Kle
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Figure 3.40: SEM micrographs of Kle
Page 189 and 190:
C D 189
Page 191 and 192:
Page 193 and 194:
Page 195 and 196:
3.5.2.5 SEM of Citrobacter For Citr
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Figure 3.47: SEM micrographs of Cit
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Figure 3.49: SEM micrographs of Cit
Page 201 and 202:
Page 203 and 204:
Table 3.26: Comparison of changes i
Page 205 and 206:
Figure 3.54: Transmission micrograp
Page 207 and 208:
Page 209 and 210:
3.5.2.2. TEM for E.coli: TEM images
Page 211 and 212:
Page 213 and 214:
Page 215 and 216:
Figure 3.62: 2-D protein electropho
Page 217 and 218:
“The therapeutic potential of unc
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Non-peroxide honey shown to be effe
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Sugars are the major content of hon
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Acacia (thorn trees), Myrtacae euca
Page 225 and 226:
major wound pathogens, consistently
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MIC % (w/v) Previous studies MIC %
Page 229 and 230:
Figure 4.3: Mean MIC and MBC (%w/v)
Page 231 and 232:
4.4 Inhibition of test organisms by
Page 233 and 234:
honey chemistry. Nevertheless, obse
Page 235 and 236:
In TEM, the cell membrane was inter
Page 237 and 238:
with more than (3-log10 reduction).
Page 239 and 240:
To date there is little research on
Page 241 and 242:
The effect of defensin peptide was
Page 243 and 244:
4.6 Further investigations: There i
Page 245 and 246:
Chapter 5 References 245
Page 247 and 248:
� Al-Jabri, A. A., Nzeako, B. C.,
Page 249 and 250:
� Bang, L. M., Buntting, C. and M
Page 251 and 252:
� Bou, G., Oliver, A. and Martine
Page 253 and 254:
� Cooper, R. (2005) The Antimicro
Page 255 and 256:
� Efem, S. E., Udoh, K. T. and Iw
Page 257 and 258:
� Greenwood, D., Finch, P., Davey
Page 259 and 260:
� Kagan, B.L., Ganz, T. and Lehre
Page 261 and 262:
esistant Staphylococcus aureus (CA-
Page 263 and 264:
� Murray, P. R., Baron, E. J., Jo
Page 265 and 266:
� Scholar, E. M. and Pratt, W. B.
Page 267 and 268:
� Subrahmanyam, M. (1998) A prosp
Page 269 and 270:
� Wadi, M., Al-Amin, H., Farouq,
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