臺灣博碩士論文加值系統

Acinetobacter baumannii associated nosocomial infections emerged as research question during the past few decades. It provides challenging opportunities to find alternative treatment option for A. baumannii to all medical researchers. The alternative to antibiotics treatment, bacteriophage therapy is a promising option for multi-drug resistant (MDR) bacterial infections. A. baumannii M3237 and 54149 clinical strains are one of the hosts of lytic phage ϕAB2 and ϕAB6, respectively. So far, depolymerase activity of tail fiber protein derived from lytic phage TFPϕAB6 has reported for hydrolyzing the exopolysaccharides of A. baumannii 54149 either by cell infection or lysis. Here, we have focused on the prevention and eradication of biofilm forming capacity of phage ϕAB6 derived tail fiber protein TFPϕAB6. The results showed that TFPϕAB6 have biofilms eradication effects on A. baumannii 54149 under the optical microscope. Secondly, the electron microscopy observation demonstrated that TFPϕAB6 cause extracellular changes of A. baumannii 54149. The similar types of effects have observed on the silicon-coated Foley catheter as the medical device. Atomic force microscopic analysis also provides more insight about TFPϕAB6 effects on morphological change of A. baumannii 54149. Thirdly, TFPϕAB6 have shown a significant difference in the colony counting of A. baumannii 54149 viable cells. Live/dead BacLight screening have confirmed the cell lysis function of TFPϕAB6 whereas, confocal laser scanning microscopy study explained the biofilm thickness reduction. Fourthly, Trapping and near-infared laser based photothermal killing function of TF2-Fe3O4@Al2O3 MNPs and TF6-Fe3O4@Al2O3 MNPs nanoprobes have demonstrated the cell lysis effects of A. baumannii M3237 and 54149 in electron microscopy. The TF2-Fe3O4@Al2O3 MNPs and TF6-Fe3O4@Al2O3 trapping based photothermal killing effects significantly reduced the colony counting of A. baumannii M3237 and 54149 viable cells. In vivo assessment of possible TFPϕAB6 therapy have shown the efficacy in a zebrafish infection model. Finally, the findings provide the insight of TFPϕAB6 have become an innovative applicable approach to detect and control medical device associated biofilm formation of A. baumannii. Altogether, TFPϕAB2 and TFPϕAB6 are good alternative candidates to overcome the MDR problem of A. baumannii.

Acinetobacter baumannii associated nosocomial infections emerged as research question during the past few decades. It provides challenging opportunities to find alternative treatment option for A. baumannii to all medical researchers. The alternative to antibiotics treatment, bacteriophage therapy is a promising option for multi-drug resistant (MDR) bacterial infections. A. baumannii M3237 and 54149 clinical strains are one of the hosts of lytic phage ϕAB2 and ϕAB6, respectively. So far, depolymerase activity of tail fiber protein derived from lytic phage TFPϕAB6 has reported for hydrolyzing the exopolysaccharides of A. baumannii 54149 either by cell infection or lysis. Here, we have focused on the prevention and eradication of biofilm forming capacity of phage ϕAB6 derived tail fiber protein TFPϕAB6. The results showed that TFPϕAB6 have biofilms eradication effects on A. baumannii 54149 under the optical microscope. Secondly, the electron microscopy observation demonstrated that TFPϕAB6 cause extracellular changes of A. baumannii 54149. The similar types of effects have observed on the silicon-coated Foley catheter as the medical device. Atomic force microscopic analysis also provides more insight about TFPϕAB6 effects on morphological change of A. baumannii 54149. Thirdly, TFPϕAB6 have shown a significant difference in the colony counting of A. baumannii 54149 viable cells. Live/dead BacLight screening have confirmed the cell lysis function of TFPϕAB6 whereas, confocal laser scanning microscopy study explained the biofilm thickness reduction. Fourthly, Trapping and near-infared laser based photothermal killing function of TF2-Fe3O4@Al2O3 MNPs and TF6-Fe3O4@Al2O3 MNPs nanoprobes have demonstrated the cell lysis effects of A. baumannii M3237 and 54149 in electron microscopy. The TF2-Fe3O4@Al2O3 MNPs and TF6-Fe3O4@Al2O3 trapping based photothermal killing effects significantly reduced the colony counting of A. baumannii M3237 and 54149 viable cells. In vivo assessment of possible TFPϕAB6 therapy have shown the efficacy in a zebrafish infection model. Finally, the findings provide the insight of TFPϕAB6 have become an innovative applicable approach to detect and control medical device associated biofilm formation of A. baumannii. Altogether, TFPϕAB2 and TFPϕAB6 are good alternative candidates to overcome the MDR problem of A. baumannii.

CHAPTER 1 6
INTRODUCTION 6
1.1. Acinetobacter baumannii 6
1.2. Antibiotic resistance of A. baumannii 6
1.3. Phage depolymerases 7
1.4. Biofilms 9
1.5. Medical devices associated biofilm formation 10
1.6. Nanoparticle technology 10
1.7. Zebrafish animal model 11
1.8. Perspective of research goal 12
CHAPTER 2 14
MATERIALS AND METHODS 14
2.1. Bacterial strains and culture conditions 14
2.2. Phage preparation of high-titer phage lysates 14
2.3. Plaque forming assay 15
2.4. Protein expression and purification 15
2.5. Spot test of tail fiber proteins (TFPϕAB2 and TFPϕAB6) 16
2.6. Minimum biofilm inhibitory concentration (MBIC) 16
2.7. Cell viability test 17
2.8. Growth curve 18
2.9. Biofilm formation assay 18
2.10. Biofilm degradation assay 19
2.11. Biofilm prevention assay 19
2.12. Biofilm inhibition assay 20
2.13. Biofilm reduction assay 20
2.14. Light microscopic observation 21
2.15. Scanning Electron Microscopy 21
2.16. Transmission electron microscopy 22
2.17. Atomic force microscopy 22
2.18. Fluorescence microscopy 23
2.19. Confocal laser scanning microscopy 24
2.20. In vivo survival study in Zebrafish 24

CHAPTER 3 25
RESULTS 25
3.1. Confirmation of ϕAB6 and ϕAB2 recombinant tail fiber protein molecular size 25
3.2. Spot test of TFPϕAB6 and TFPϕAB2 makes a semi-clear zone on Ab-54149 and Ab-M3237 25
3.3. Minimum biofilm inhibitory concentration (MBIC) of TFPϕAB6 against Ab-54149 26
3.4. The effect of TFPϕAB6 on Ab-54149 cell viability 26
3.5. TFPϕAB6 slightly reduce Ab-54149 growth curve 27
3.6. Biofilm forming ability of clinical isolates of Acinetobacter baumannii 27
3.7. Biofilm degradation efficacy of TFPϕAB6 against Ab-54149 28
3.8. Determination of biofilm prevention efficiency of TFPϕAB6 28
3.9. TFPϕAB6 influence biofilm inhibition of Ab-54149 29
3.10. Ab-54149 biofilm reduction impact by TFPϕAB6 29
3.11. Biofilm eradication assessment of TFPϕAB6 under light microscope 30
3.12. TFPϕAB2 and TFPϕAB6 caused cell morphology change of A. baumannii strains 30
3.13. Biofilm prevention and degradation activity of TFPϕAB6 on medical device 31
3.14. Observation of cell lysis effect of TFPϕAB6 in Transmission electron microscope 32
3.15. AFM images of Ab-54149 cell surface change treated with TFPϕAB6 32
3.16. Live/Dead BacLight screening of Ab-54149 by TFPϕAB6 caused cell damaged 33
3.17. Reduction of Ab-54149 biofilm thickness by TFPϕAB6 33
3.18. In vivo treatment of TFPϕAB6 against A. baumannii infection 34
3.19. TEM trapping capacity of TFPs-Fe3O4@Al2O3 MNPs on A. baumannii strains 34
3.20. TFPs-Fe3O4@Al2O3 MNPs enhanced photothermal based killing of A. baumannii 35
CHAPTER 4 36
DISCUSSION AND CONCLUSION 36
References 42
Appendixes 49

1Peleg, A. Y., Seifert, H. & Paterson, D. L. Acinetobacter baumannii: emergence of a successful pathogen. Clinical microbiology reviews 21, 538-582 (2008).
2Qi, L. et al. Relationship between antibiotic resistance, biofilm formation, and biofilm-specific resistance in Acinetobacter baumannii. Frontiers in microbiology 7, 483 (2016).
3Bergogne-Berezin, E. & Towner, K. Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features. Clinical microbiology reviews 9, 148 (1996).
4Patwardhan, R., Dhakephalkar, P., Niphadkar, K. & Chopade, B. A study on nosocomial pathogens in ICU with special reference to multiresistant Acinetobacter baumannii harbouring multiple plasmids. Indian Journal of Medical Research 128, 178 (2008).
5Su, C.-H. et al. Increase of carbapenem-resistant Acinetobacter baumannii infection in acute care hospitals in Taiwan: association with hospital antimicrobial usage. PloS one 7, e37788 (2012).
6Kempf, M. & Rolain, J.-M. Emergence of resistance to carbapenems in Acinetobacter baumannii in Europe: clinical impact and therapeutic options. International journal of antimicrobial agents 39, 105-114 (2012).
7Diancourt, L., Passet, V., Nemec, A., Dijkshoorn, L. & Brisse, S. The population structure of Acinetobacter baumannii: expanding multiresistant clones from an ancestral susceptible genetic pool. PloS one 5, e10034 (2010).
8Perez, F. et al. Global challenge of multidrug-resistant Acinetobacter baumannii. Antimicrobial agents and chemotherapy 51, 3471-3484 (2007).
9Lai, M.-J. et al. Identification and characterisation of the putative phage-related endolysins through full genome sequence analysis in Acinetobacter baumannii ATCC 17978. International journal of antimicrobial agents 42, 141-148 (2013).
10Antunes, L. C., Imperi, F., Minandri, F. & Visca, P. In vitro and in vivo antimicrobial activity of gallium nitrate against multidrug resistant Acinetobacter baumannii. Antimicrobial agents and chemotherapy, AAC. 01519-01512 (2012).
11de Breij, A. et al. CsuA/BABCDE-dependent pili are not involved in the adherence of Acinetobacter baumannii ATCC19606T to human airway epithelial cells and their inflammatory response. Research in microbiology 160, 213-218 (2009).
12Donlan, R. M. Biofilm formation: a clinically relevant microbiological process. Clinical Infectious Diseases 33, 1387-1392 (2001).
13Manchanda, V., Sanchaita, S. & Singh, N. Multidrug resistant acinetobacter. Journal of global infectious diseases 2, 291 (2010).
14Valencia, R. et al. Nosocomial outbreak of infection with pan–drug-resistant Acinetobacter baumannii in a tertiary care university hospital. Infection Control & Hospital Epidemiology 30, 257-263 (2009).
15Montero, A. et al. Antibiotic combinations for serious infections caused by carbapenem-resistant Acinetobacter baumannii in a mouse pneumonia model. Journal of Antimicrobial Chemotherapy 54, 1085-1091 (2004).
16Costerton, J. W., Lewandowski, Z., Caldwell, D. E., Korber, D. R. & Lappin-Scott, H. M. Microbial biofilms. Annual Reviews in Microbiology 49, 711-745 (1995).
17Bentancor, L. V., O'Malley, J. M., Bozkurt-Guzel, C., Pier, G. B. & Maira-Litrán, T. Poly-N-acetyl-β-(1-6)-glucosamine is a target for protective immunity against Acinetobacter baumannii infections. Infection and immunity 80, 651-656 (2012).
18Liu, F., Aubry, A. J., Schoenhofen, I. C., Logan, S. M. & Tanner, M. E. The engineering of bacteria bearing azido‐pseudaminic acid‐modified flagella. ChemBioChem 10, 1317-1320 (2009).
19Asif, M., Alvi, I. A. & Rehman, S. U. Insight into Acinetobacter baumannii: pathogenesis, global resistance, mechanisms of resistance, treatment options, and alternative modalities. Infection and drug resistance 11, 1249 (2018).
20Zhao, X. et al. Outer membrane proteins ail and OmpF of Yersinia pestis are involved in the adsorption of T7-related bacteriophage Yep-phi. Journal of virology, JVI. 01948-01913 (2013).
21Scholl, D., Rogers, S., Adhya, S. & Merril, C. R. Bacteriophage K1-5 encodes two different tail fiber proteins, allowing it to infect and replicate on both K1 and K5 strains of Escherichia coli. Journal of virology 75, 2509-2515 (2001).
22Hughes, K., Sutherland, I., Clark, J. & Jones, M. Bacteriophage and associated polysaccharide depolymerases–novel tools for study of bacterial biofilms. Journal of applied microbiology 85, 583-590 (1998).
23Pires, D. P., Oliveira, H., Melo, L. D., Sillankorva, S. & Azeredo, J. Bacteriophage-encoded depolymerases: their diversity and biotechnological applications. Applied microbiology and biotechnology 100, 2141-2151 (2016).
24Cornelissen, A. et al. Identification of EPS-degrading activity within the tail spikes of the novel Pseudomonas putida phage AF. Virology 434, 251-256 (2012).
25Casjens, S. R. & Molineux, I. J. in Viral Molecular Machines 143-179 (Springer, 2012).
26Dowah, A. S. & Clokie, M. R. Review of the nature, diversity and structure of bacteriophage receptor binding proteins that target Gram-positive bacteria. Biophysical reviews, 1-8 (2018).
27Yan, J., Mao, J. & Xie, J. Bacteriophage polysaccharide depolymerases and biomedical applications. BioDrugs 28, 265-274 (2014).
28Cornelissen, A. et al. The T7-related Pseudomonas putida phage φ15 displays virion-associated biofilm degradation properties. PLoS One 6, e18597 (2011).
29Gutiérrez, D., Martínez, B., Rodríguez, A. & García, P. Genomic characterization of two Staphylococcus epidermidis bacteriophages with anti-biofilm potential. BMC genomics 13, 228 (2012).
30Tait, K., Skillman, L. & Sutherland, I. The efficacy of bacteriophage as a method of biofilm eradication. Biofouling 18, 305-311 (2002).
31Lu, T. K. & Collins, J. J. Dispersing biofilms with engineered enzymatic bacteriophage. Proceedings of the National Academy of Sciences 104, 11197-11202 (2007).
32Lin, T.-L. et al. Isolation of a bacteriophage and its depolymerase specific for K1 capsule of Klebsiella pneumoniae: implication in typing and treatment. The Journal of infectious diseases 210, 1734-1744 (2014).
33Ceyssens, P.-J. et al. Genomic analysis of Pseudomonas aeruginosa phages LKD16 and LKA1: Establishment of the φKMV subgroup within the T7 supergroup. Journal of bacteriology 188, 6924-6931 (2006).
34Thompson, J. E. et al. The K5 lyase KflA combines a viral tail spike structure with a bacterial polysaccharide lyase mechanism. Journal of Biological Chemistry, jbc. M110. 127571 (2010).
35Majkowska-Skrobek, G. et al. Capsule-targeting depolymerase, derived from Klebsiella KP36 phage, as a tool for the development of anti-virulent strategy. Viruses 8, 324 (2016).
36Sutherland, I. W. Polysaccharide lyases. FEMS microbiology reviews 16, 323-347 (1995).
37Donlan, R. M. & Costerton, J. W. Biofilms: survival mechanisms of clinically relevant microorganisms. Clinical microbiology reviews 15, 167-193 (2002).
38Choi, A. H., Slamti, L., Avci, F. Y., Pier, G. B. & Maira-Litrán, T. The pgaABCD locus of Acinetobacter baumannii encodes the production of poly-β-1-6-N-acetylglucosamine, which is critical for biofilm formation. Journal of bacteriology 191, 5953-5963 (2009).
39Xu, G., Ryan, C., Kiefel, M. J., Wilson, J. C. & Taylor, G. L. Structural studies on the Pseudomonas aeruginosa sialidase-like enzyme PA2794 suggest substrate and mechanistic variations. Journal of molecular biology 386, 828-840 (2009).
40Howard, S. L. et al. Campylobacter jejuni glycosylation island important in cell charge, legionaminic acid biosynthesis, and colonization of chickens. Infection and immunity 77, 2544-2556 (2009).
41Kao, C.-Y., Sheu, B.-S. & Wu, J.-J. Helicobacter pylori infection: An overview of bacterial virulence factors and pathogenesis. Biomedical journal 39, 14-23 (2016).
42Lee, I.-M. et al. Pseudaminic Acid on Exopolysaccharide of Acinetobacter baumannii Plays a Critical Role in Phage-Assisted Preparation of Glycoconjugate Vaccine with High Antigenicity. Journal of the American Chemical Society 140, 8639-8643 (2018).
43Fux, C., Wilson, S. & Stoodley, P. Detachment characteristics and oxacillin resistance of Staphyloccocus aureus biofilm emboli in an in vitro catheter infection model. Journal of bacteriology 186, 4486-4491 (2004).
44Raad, I. I. et al. Vancomycin-resistant Enterococcus faecium: catheter colonization, esp gene, and decreased susceptibility to antibiotics in biofilm. Antimicrobial agents and chemotherapy 49, 5046-5050 (2005).
45Curtin, J. J. & Donlan, R. M. Using bacteriophages to reduce formation of catheter-associated biofilms by Staphylococcus epidermidis. Antimicrobial agents and chemotherapy 50, 1268-1275 (2006).
46Donlan, R. M. Biofilms and device-associated infections. Emerging infectious diseases 7, 277 (2001).
47Jacobsen, S. á., Stickler, D., Mobley, H. & Shirtliff, M. Complicated catheter-associated urinary tract infections due to Escherichia coli and Proteus mirabilis. Clinical microbiology reviews 21, 26-59 (2008).
48Milo, S. et al. Prevention of encrustation and blockage of urinary catheters by Proteus mirabilis via pH-triggered release of bacteriophage. Journal of Materials Chemistry B 5, 5403-5411 (2017).
49Cole, S. J., Records, A. R., Orr, M. W., Linden, S. B. & Lee, V. T. Catheter-associated urinary tract infection by Pseudomonas aeruginosa is mediated by exopolysaccharide independent biofilms. Infection and immunity, IAI. 01652-01614 (2014).
50Campana, R., Casettari, L., Ciandrini, E., Illum, L. & Baffone, W. Chitosans inhibit the growth and the adhesion of Klebsiella pneumoniae and Escherichia coli clinical isolates on urinary catheters. International journal of antimicrobial agents 50, 135-141 (2017).
51Rodríguez‐Baño, J. et al. Biofilm formation in Acinetobacter baumannii: associated features and clinical implications. Clinical microbiology and infection 14, 276-278 (2008).
52Liu, H.-L. & Yang, T. C.-K. Photocatalytic inactivation of Escherichia coli and Lactobacillus helveticus by ZnO and TiO2 activated with ultraviolet light. Process Biochemistry 39, 475-481 (2003).
53Zhang, L., Jiang, Y., Ding, Y., Povey, M. & York, D. Investigation into the antibacterial behaviour of suspensions of ZnO nanoparticles (ZnO nanofluids). Journal of Nanoparticle Research 9, 479-489 (2007).
54Stoimenov, P. K., Klinger, R. L., Marchin, G. L. & Klabunde, K. J. Metal oxide nanoparticles as bactericidal agents. Langmuir 18, 6679-6686 (2002).
55Sawai, J. Quantitative evaluation of antibacterial activities of metallic oxide powders (ZnO, MgO and CaO) by conductimetric assay. Journal of microbiological methods 54, 177-182 (2003).
56Ren, G. et al. Characterisation of copper oxide nanoparticles for antimicrobial applications. International journal of antimicrobial agents 33, 587-590 (2009).
57García-Saucedo, C., Field, J. A., Otero-Gonzalez, L. & Sierra-Álvarez, R. Low toxicity of HfO2, SiO2, Al2O3 and CeO2 nanoparticles to the yeast, Saccharomyces cerevisiae. Journal of hazardous materials 192, 1572-1579 (2011).
58Thill, A. et al. Cytotoxicity of CeO2 Nanoparticles for Escherichia coli. Physico-Chemical Insight of the Cytotoxicity Mechanism. Environmental Science & Technology 40, 6151-6156, doi:10.1021/es060999b (2006).
59Brayner, R. et al. Toxicological Impact Studies Based on Escherichia coli Bacteria in Ultrafine ZnO Nanoparticles Colloidal Medium. Nano Letters 6, 866-870, doi:10.1021/nl052326h (2006).
60Kreyling, W. G., Semmler-Behnke, M. & Chaudhry, Q. A complementary definition of nanomaterial. Nano today 5, 165-168 (2010).
61Hameed, A. S. H. et al. In vitro antibacterial activity of ZnO and Nd doped ZnO nanoparticles against ESBL producing Escherichia coli and Klebsiella pneumoniae. Scientific reports 6, 24312 (2016).
62Ma, C. et al. Bi-phase dispersible Fe3O4@ Au core–shell multifunctional nanoparticles: synthesis, characterization and properties. Composite Interfaces, 1-13 (2018).
63Huang, S.-Y. & Chen, Y.-C. Magnetic nanoparticle-based platform for characterization of histidine-rich proteins and peptides. Analytical chemistry 85, 3347-3354 (2013).
64Xu, C. et al. Dopamine as a robust anchor to immobilize functional molecules on the iron oxide shell of magnetic nanoparticles. Journal of the American Chemical Society 126, 9938-9939 (2004).
65Lin, J.-Y. & Chen, Y.-C. Functional magnetic nanoparticle-based trapping and sensing approaches for label-free fluorescence detection of DNA. Talanta 86, 200-207 (2011).
66Lo, C.-Y., Chen, W.-Y., Chen, C.-T. & Chen, Y.-C. Rapid enrichment of phosphopeptides from tryptic digests of proteins using iron oxide nanocomposites of magnetic particles coated with zirconia as the concentrating probes. Journal of proteome research 6, 887-893 (2007).
67Chen g-Tai Chen et al. Rapid enrichment of phosphopeptides and phosphoproteins from complex samples using magnetic particles coated with alumina as the concentrating probes for MALDI MS analysis. Journal of proteome research 6, 316-325 (2006).
68Chen, C.-T. & Chen, Y.-C. Trapping performance of Fe3O4@ Al2O3 and Fe3O4@ TiO2 magnetic nanoparticles in the selective enrichment of phosphopeptides from human serum. Journal of Biomedical Nanotechnology 4, 73-79 (2008).
69Yu, T.-J., Li, P.-H., Tseng, T.-W. & Chen, Y.-C. Multifunctional Fe3O4/alumina core/shell MNPs as photothermal agents for targeted hyperthermia of nosocomial and antibiotic-resistant bacteria. Nanomedicine 6, 1353-1363 (2011).
70Chen, W.-J. & Chen, Y.-C. Fe3O4/TiO2 core/shell magnetic nanoparticle-based photokilling of pathogenic bacteria. Nanomedicine 5, 1585-1593 (2010).
71Westerfield, M. The zebrafish book: a guide for the laboratory use of zebrafish. http://zfin. org/zf_info/zfbook/zfbk. html (2000).
72Herbomel, P., Thisse, B. & Thisse, C. Ontogeny and behaviour of early macrophages in the zebrafish embryo. Development 126, 3735-3745 (1999).
73Davis, J. M. et al. Real-time visualization of mycobacterium-macrophage interactions leading to initiation of granuloma formation in zebrafish embryos. Immunity 17, 693-702 (2002).
74Meijer, A. H. et al. Transcriptome profiling of adult zebrafish at the late stage of chronic tuberculosis due to Mycobacterium marinum infection. Molecular immunology 42, 1185-1203 (2005).
75Lieschke, G. J. & Currie, P. D. Animal models of human disease: zebrafish swim into view. Nature Reviews Genetics 8, 353 (2007).
76Yang, H., Liang, L., Lin, S. & Jia, S. Isolation and characterization of a virulent bacteriophage AB1 of Acinetobacter baumannii. BMC microbiology 10, 131 (2010).
77Shankar, R. et al. A novel antibacterial gene transfer treatment for multidrug-resistant Acinetobacter baumannii-induced burn sepsis. Journal of burn care & research 28, 6-12 (2007).
78McConnell, M. J. et al. Vaccination with outer membrane complexes elicits rapid protective immunity to multidrug-resistant Acinetobacter baumannii. Infection and immunity 79, 518-526 (2011).
79Tsai, T. et al. Chitosan augments photodynamic inactivation of gram-positive and gram-negative bacteria. Antimicrobial agents and chemotherapy (2011).
80Mihu, M. R. et al. The use of nitric oxide releasing nanoparticles as a treatment against Acinetobacter baumannii in wound infections. Virulence 1, 62-67 (2010).
81Nomura, M., Hall, B. D. & Spiegelman, S. Characterization of RNA synthesized in Escherichia coli after bacteriophage T2 infection. Journal of Molecular Biology 2, 306-IN304 (1960).
82De Siqueira, R., Dodd, C. & Rees, C. Evaluation of the natural virucidal activity of teas for use in the phage amplification assay. International journal of food microbiology 111, 259-262 (2006).
83Kropinski, A. M., Mazzocco, A., Waddell, T. E., Lingohr, E. & Johnson, R. P. in Bacteriophages 69-76 (Springer, 2009).
84Lillehaug, D. An improved plaque assay for poor plaque‐producing temperate lactococcal bacteriophages. Journal of applied microbiology 83, 85-90 (1997).
85Hathaway, H. et al. Thermally triggered release of the bacteriophage endolysin CHAPK and the bacteriocin lysostaphin for the control of methicillin resistant Staphylococcus aureus (MRSA). Journal of Controlled Release 245, 108-115 (2017).
86Tiller, J. C., Liao, C.-J., Lewis, K. & Klibanov, A. M. Designing surfaces that kill bacteria on contact. Proceedings of the National Academy of Sciences 98, 5981-5985 (2001).
87Selasi, G. N. et al. Differences in biofilm mass, expression of biofilm-associated genes, and resistance to desiccation between epidemic and sporadic clones of carbapenem-resistant Acinetobacter baumannii sequence Type 191. PloS one 11, e0162576 (2016).
88Sasikala, D. & Srinivasan, P. Characterization of potential lytic bacteriophage against Vibrio alginolyticus and its therapeutic implications on biofilm dispersal. Microbial pathogenesis 101, 24-35 (2016).
89Schooley, R. T. et al. Development and use of personalized bacteriophage-based therapeutic cocktails to treat a patient with a disseminated resistant Acinetobacter baumannii infection. Antimicrobial agents and chemotherapy, AAC. 00954-00917 (2017).
90Son, J.-S. et al. Antibacterial and biofilm removal activity of a podoviridae Staphylococcus aureus bacteriophage SAP-2 and a derived recombinant cell-wall-degrading enzyme. Applied microbiology and biotechnology 86, 1439-1449 (2010).
91Peng, S.-Y. et al. Highly potent antimicrobial modified peptides derived from the Acinetobacter baumannii phage endolysin LysAB2. Scientific reports 7, 11477 (2017).
92Liou, J.-W. et al. Visible light responsive photocatalyst induces progressive and apical-terminus preferential damages on Escherichia coli surfaces. PloS one 6, e19982 (2011).
93Zhang, Y. & Hu, Z. Combined treatment of Pseudomonas aeruginosa biofilms with bacteriophages and chlorine. Biotechnology and bioengineering 110, 286-295 (2013).
94Runci, F., Bonchi, C., Frangipani, E., Visaggio, D. & Visca, P. Acinetobacter baumannii biofilm formation in Human serum and disruption by gallium. Antimicrobial agents and chemotherapy 61, e01563-01516 (2017).
95Lai, M.-J. et al. The tail associated protein of Acinetobacter baumannii phage ΦAB6 is the host specificity determinant possessing exopolysaccharide depolymerase activity. PloS one 11, e0153361 (2016).
96Hsieh, P.-F., Lin, H.-H., Lin, T.-L., Chen, Y.-Y. & Wang, J.-T. Two T7-like bacteriophages, K5-2 and K5-4, each encodes two capsule depolymerases: isolation and functional characterization. Scientific reports 7, 4624 (2017).
97Oliveira, H. et al. Ability of phages to infect Acinetobacter calcoaceticus‐Acinetobacter baumannii complex species through acquisition of different pectate lyase depolymerase domains. Environmental microbiology 19, 5060-5077 (2017).
98Becker, S. C. et al. Triple-acting lytic enzyme treatment of drug-resistant and intracellular Staphylococcus aureus. Scientific reports 6, 25063 (2016).
99Lai, C.-C. et al. Implementation of a national bundle care program to reduce catheter-associated urinary tract infection in high-risk units of hospitals in Taiwan. Journal of Microbiology, Immunology and Infection 50, 464-470 (2017).
100Oliveira, H. et al. Functional analysis and anti-virulent properties of a new depolymerase from a myovirus that infects Acinetobacter baumannii capsule K45. Journal of virology, JVI. 01163-01118 (2018).
101Glonti, T., Chanishvili, N. & Taylor, P. Bacteriophage‐derived enzyme that depolymerizes the alginic acid capsule associated with cystic fibrosis isolates of Pseudomonas aeruginosa. Journal of applied microbiology 108, 695-702 (2010).
102Xiang, Y. et al. Crystal and cryoEM structural studies of a cell wall degrading enzyme in the bacteriophage φ29 tail. Proceedings of the National Academy of Sciences 105, 9552-9557 (2008).
103Rashel, M. et al. Tail-associated structural protein gp61 of Staphylococcus aureus phage ΦMR11 has bifunctional lytic activity. FEMS microbiology letters 284, 9-16 (2008).
104Kenny, J. G., McGrath, S., Fitzgerald, G. F. & van Sinderen, D. Bacteriophage Tuc2009 encodes a tail-associated cell wall-degrading activity. Journal of bacteriology 186, 3480-3491 (2004).
105Kasas, S., Fellay, B. & Cargnello, R. Observation of the action of penicillin on Bacillus subtilis using atomic force microscopy: technique for the preparation of bacteria. Surface and interface analysis 21, 400-401 (1994).
106Amro, N. A. et al. High-resolution atomic force microscopy studies of the Escherichia coli outer membrane: structural basis for permeability. Langmuir 16, 2789-2796 (2000).
107Liou, J.-W., Hung, Y.-J., Yang, C.-H. & Chen, Y.-C. The antimicrobial activity of gramicidin A is associated with hydroxyl radical formation. PloS one 10, e0117065 (2015).
108Wolfaardt, G. M., Lawrence, J. R. & Korber, D. R. in Microbial extracellular polymeric substances 171-200 (Springer, 1999).
109Silva, Y. J. et al. Phage therapy as an approach to prevent Vibrio anguillarum infections in fish larvae production. PLoS One 9, e114197 (2014).
110Díez-Martínez, R. et al. Improved Lethal Effect of Cpl-7, a Pneumococcal Phage Lysozyme of Broad Bactericidal Activity by Inverting Net Charge of its Cell Wall-Binding Module. Antimicrobial agents and chemotherapy, AAC. 01372-01313 (2013).
111Hooton, S. P., Timms, A. R., Rowsell, J., Wilson, R. & Connerton, I. F. Salmonella Typhimurium-specific bacteriophage ΦSH19 and the origins of species specificity in the Vi01-like phage family. Virology journal 8, 498 (2011).
112Pickard, D. et al. A conserved acetyl esterase domain targets diverse bacteriophages to the Vi capsular receptor of Salmonella enterica serovar Typhi. Journal of bacteriology 192, 5746-5754 (2010).