Effect of N-Acetylcysteine in Combination with Antibiotics on the Biofilms of Three Cystic Fibrosis Pathogens of Emerging Importance
Abstract
:1. Introduction
2. Results
2.1. Neutralised NAC Is Ineffective against Planktonic Bacterial Growth
2.2. Effect of Antibiotics against Planktonic Bacterial Growth
2.3. Combination Therapy Demonstrated Synergism and Additive Effect against Planktonic Bacteria in Checkerboard Assays
2.4. Effect of Synergistic and Additive Combination Therapies against Irreversibly Attached Bacteria
2.5. Time Course Bacteriostatic Effect of Synergistic and Additive Combination Therapies
2.6. Effect of Synergistic and Additive Combination Therapies against Mature Biofilms In Vitro
3. Discussion
4. Materials and Methods
4.1. Cystic Fibrosis Opportunist Strains Tested
4.2. Culture Conditions and Preparations of Treatments
4.3. Preparation of NAC Medium
4.4. Preparation of Antibiotics
4.5. Effect of Antioxidant and Antibiotics against PLANKTONIC Bacteria
4.6. Synergy Testing via Checkerboard Assays
4.7. Effect of Antioxidant and Antibiotics on Prevention of Biofilm Maturation Following Irreversible Bacterial Attachment
4.8. Time-Kill Assay of Single versus Combination Treatments against Planktonic Cultures
4.9. In Vitro Mature Biofilm Susceptibility Testing with Single and Combination Treatments Using Colony Forming Units (CFU/mL)
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Bacteria | Strain | Source | Antibiotic Profile | ||||
---|---|---|---|---|---|---|---|
TZP110 | CAZ30 | MEM10 | CIP5 | SXT25 | |||
B. cenocepacia | ATCC 25608TM | ATCC (Manassas, Virginia) (Incision wound) | NA | S | S | NA | S |
M2167 + | CF Sputum (+) | NA | R | R | NA | R | |
E5452 + | CF Sputum (+) | NA | S | R | NA | R | |
E5328 + | CF Sputum (+) | NA | S | S | NA | R | |
S. maltophilia | ATCC 13637TM | ATCC (Oropharyngeal region) | NA | NA | NA | NA | S |
445595 ^ | CF Sputum ^ | NA | NA | NA | NA | S | |
19308 ^ | CF Sputum ^ | NA | NA | NA | NA | R | |
A. xylosoxidans# | ATCC 27601TM | ATCC (Ear discharge) | S | I | S | R | R |
6268 ^ | CF Sputum ^ | S | I | S | R | R | |
6908 ^ | CF Sputum ^ | S | I | S | R | R | |
4365 ^ | CF Sputum ^ | NA | NA | NA | NA | R |
Bacteria | Neutral NAC (mg/mL) | Ciprofloxacin (μg/mL) | Colistin (μg/mL) | Tobramycin (μg/mL) | |
---|---|---|---|---|---|
A. xylosoxidans | ATCC 27601TM | >16.3 | 32 | >128 | >128 |
6268 | >16.3 | 32 | 32 | >128 | |
6908 | >16.3 | 32 | 8 | >128 | |
4365 | >16.3 | 32 | >128 | >128 | |
B. cenocepacia | ATCC 25608 TM | >16.3 | 64 | 16 | >128 |
M2167 | >16.3 | 64 | >128 | >128 | |
E5452 | >16.3 | 64 | 64 | >128 | |
E5328 | >16.3 | 64 | >128 | >128 | |
S. maltophilia | ATCC 13637 TM | >16.3 | 16 | 32 | 64 |
445595 | >16.3 | 64 | 64 | >128 | |
19308 | >16.3 | 32 | 64 | 64 |
Letter Reference | Neutral NAC (mg/mL) | Antibiotic (μg/mL) |
---|---|---|
A | 2 | 32 μg/mL Ciprofloxacin |
B | 4.1 | |
C | 8.2 | |
D | 1 | 64 μg/mL Ciprofloxacin |
E | 2 | 8 μg/mL Colistin |
F | 4.1 | |
G | 8.2 | |
H | 2 | 16 μg/mL Colistin |
I | 4.1 | |
J | 8.2 | |
K | 16.3 | |
L | 16.3 | 32 μg/mL Colistin |
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Aiyer, A.; Visser, S.K.; Bye, P.; Britton, W.J.; Whiteley, G.S.; Glasbey, T.; Kriel, F.H.; Farrell, J.; Das, T.; Manos, J. Effect of N-Acetylcysteine in Combination with Antibiotics on the Biofilms of Three Cystic Fibrosis Pathogens of Emerging Importance. Antibiotics 2021, 10, 1176. https://doi.org/10.3390/antibiotics10101176
Aiyer A, Visser SK, Bye P, Britton WJ, Whiteley GS, Glasbey T, Kriel FH, Farrell J, Das T, Manos J. Effect of N-Acetylcysteine in Combination with Antibiotics on the Biofilms of Three Cystic Fibrosis Pathogens of Emerging Importance. Antibiotics. 2021; 10(10):1176. https://doi.org/10.3390/antibiotics10101176
Chicago/Turabian StyleAiyer, Aditi, Simone K. Visser, Peter Bye, Warwick J. Britton, Gregory S. Whiteley, Trevor Glasbey, Frederik H. Kriel, Jessica Farrell, Theerthankar Das, and Jim Manos. 2021. "Effect of N-Acetylcysteine in Combination with Antibiotics on the Biofilms of Three Cystic Fibrosis Pathogens of Emerging Importance" Antibiotics 10, no. 10: 1176. https://doi.org/10.3390/antibiotics10101176