Inactivation of Polymicrobial Biofilms of Foodborne Pathogens Using Epsilon Poly-L-Lysin Conjugated Chitosan Nanoparticles
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial and Mammalian Cell Lines Used in This Study
2.2. Synthesis of Chitosan Nanoparticles and Conjugation with ε-poly-L-lysine
2.3. Antibacterial Activity Testing of Chitosan Nanoparticles
2.4. Cell Proliferation and Cytotoxicity Tests of ChNP-PL
2.5. Single and Mixed Culture Biofilm Formation
2.6. Prevention of Biofilm Formation and Inactivation of Preformed Biofilm by ChNP-PL
3. Results
3.1. Synthesis of Chitosan Nanoparticles Conjugated with ε-poly-L-lysine (ChNP-PL)
3.2. ChNP and PL Exhibited Synergistic Antimicrobial Activity
3.3. ChNP-PL Is Nontoxic to Intestinal Epithelial Cells
3.4. Shifting of Population Dynamics within Mixed Culture Biofilms
3.5. ChNP-PL Effectively Prevented Biofilm Formation by Mono- or Mixed-Cultures
3.6. ChNP-PL Inactivated Preformed Biofilms by All Tested Bacteria except P. aeruginosa
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Bacteria | MIC (µg/mL) | |
---|---|---|
ChNP | ChNP-PL | |
Pseudomonas aeruginosa ATCC10145 | >37.5 | 12.5–25 |
P. putida PRI107 | >37.5 | 12.5–25 |
P. aeruginosa PRI99 | 25–37.5 | 2.5–3.75 |
Listeria ivanovii ATCC19119 | 25–37.5 | 2.5–3.75 |
L. seeligeri ATCC 35967 | 25–37.5 | 2.5–3.75 |
L. marthii ATCC BAA-1595 | 25–37.5 | 2.5–3.75 |
L. monocytogenes F40 | 25–37.5 | 2.5–3.75 |
L. monocytogenes F4244 | 25–37.5 | 1.25–2.5 |
Salmonella enterica serovar Enteritidis PT21 | >37.5 | 3.75–12.5 |
S. enterica ser. Typhimurium ST1 | >37.5 | 12.5–25 |
S. enterica ser. Heidelberg 18ENT1418 | >37.5 | 12.5–25 |
S. enterica ser. Enteritidis 18ENT1344 | >37.5 | 3.75–12.5 |
Staphylococcus aureus NRRL B767 | >37.5 | 3.75–12.5 |
S. aureus ATCC25923 | 25–37.5 | 2.5–3.75 |
S. aureus ATCC29213 | 25–37.5 | 2.5–3.75 |
Escherichia coli K12 | >37.5 | 2.5–3.75 |
E. coli O157:H7 SEA13A72 | >37.5 | 2.5–3.75 |
E. coli O157:H7 PT23 | >37.5 | 1.25–2.5 |
E. coli O157:H7 EDL933 | >37.5 | 2.5–3.75 |
Bacteria | Avg CFU/mL | Fold-Change * | ||
---|---|---|---|---|
Initial Inoculum | Monoculture Biofilm | Mixed Culture Biofilm | ||
L. monocytogenes F4244 | 3.5 × 106 | 4.1 × 108 | 2.1 × 109 | 5-fold ↑ |
S. aureus ATCC25923 | 7.4 × 106 | 4.7 × 109 | 4.2 × 108 | 10-fold ↓ |
L. monocytogenes F4244 | 3.5 × 106 | 4.1 × 108 | 4.7 × 109 | 11-fold ↑ |
P. aeruginosa PRI99 | 7.0 × 106 | 7.3 × 109 | 4.6 × 109 | 1.6-fold ↓ |
L. monocytogenes F4244 | 3.3 × 106 | 3.2 × 108 | 1.1 × 108 | 2.9-fold ↓ |
S. aureus 2747 | 1.2 × 107 | 3.0 × 108 | 2.1 × 108 | 1.4-fold ↓ |
L. monocytogenes F4244 | 3.6 × 106 | 4.2 × 108 | 5.1 × 107 | 8.4-fold ↓ |
S. enterica serovar Enteritidis 1344 | 6.4 × 106 | 7.5 × 107 | 4.0 × 107 | 1.9-fold ↓ |
L. monocytogenes F4244 | 3.6 × 106 | 4.2 × 108 | 1.4 × 107 | 30-fold ↓ |
E. coli O157:H7 EDL933 | 3.4 × 106 | 7.6 × 107 | 3.4 × 108 | 4.5-fold ↑ |
Bacteria | Avg CFU/cm2 | ||
---|---|---|---|
Untreated | ChNP * | ChNP-PL * | |
L. monocytogenes | 9.2 × 106 | <50 (>184,000-fold ↓) | <50 (>184,000-fold ↓) |
S. aureus | 8.7 × 107 | 8.9 × 106 (9.8-fold ↓) | 3.6 × 104 (2400-fold ↓) |
P. aeruginosa | 1.7 × 107 | 5.0 × 105 (34-fold ↓) | <50 (>184,000-fold ↓) |
S. enterica ser. Enteritidis | 1.1 × 107 | 4.4 × 107 (4-fold ↑) | 103 (110,000-fold ↓) |
E. coli O157:H7 | 2.7 × 107 | 1.2 × 108 (4.4-fold ↑) | <50 (>184,000-fold ↓) |
Lm + Sa mixed biofilms | |||
L. monocytogenes | 1.9 × 107 | <50 (>184,000-fold ↓) | <50 (>184,000-fold ↓) |
S. aureus | 4.2 × 106 | 3.8 × 106 (1.1-fold ↓) | 1.0 × 104 (420-fold ↓) |
Lm + Pa mixed biofilms | |||
L. monocytogenes | 8.2 × 107 | <50 (>184,000-fold ↓) | <50 (>184,000-fold ↓) |
P. aeruginosa | 4.0 × 107 | 1.0 × 103 (40,000-fold ↓) | <50 (>184,000-fold ↓) |
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Bai, X.; Xu, L.; Singh, A.K.; Qiu, X.; Liu, M.; Abuzeid, A.; El-Khateib, T.; Bhunia, A.K. Inactivation of Polymicrobial Biofilms of Foodborne Pathogens Using Epsilon Poly-L-Lysin Conjugated Chitosan Nanoparticles. Foods 2022, 11, 569. https://doi.org/10.3390/foods11040569
Bai X, Xu L, Singh AK, Qiu X, Liu M, Abuzeid A, El-Khateib T, Bhunia AK. Inactivation of Polymicrobial Biofilms of Foodborne Pathogens Using Epsilon Poly-L-Lysin Conjugated Chitosan Nanoparticles. Foods. 2022; 11(4):569. https://doi.org/10.3390/foods11040569
Chicago/Turabian StyleBai, Xingjian, Luping Xu, Atul Kumar Singh, Xiaoling Qiu, Mai Liu, Ahmed Abuzeid, Talaat El-Khateib, and Arun K. Bhunia. 2022. "Inactivation of Polymicrobial Biofilms of Foodborne Pathogens Using Epsilon Poly-L-Lysin Conjugated Chitosan Nanoparticles" Foods 11, no. 4: 569. https://doi.org/10.3390/foods11040569