Antimicrobial and Antibiofilm Capacity of Chitosan Nanoparticles against Wild Type Strain of Pseudomonas sp. Isolated from Milk of Cows Diagnosed with Bovine Mastitis

Abstract

Bovine mastitis (BM) is the most prevalent bacterial infection in the livestock sector, affecting the dairy industry greatly. The prevention and treatment of this disease is mainly made via antibiotics, but the increasing antimicrobial resistance of pathogens has affected the efficiency of conventional drugs. Pseudomonas sp. is one of the pathogens involved in this infection. The therapeutic rate of cure for this environmental mastitis-causing pathogen is practically zero, regardless of treatment. Biofilm formation has been one of the main virulence mechanisms of Pseudomonas hence presenting resistance to antibiotic therapy. We have manufactured chitosan nanoparticles (NQo) with tripolyphosphate (TPP) using ionotropic gelation. These NQo were confronted against a Pseudomonas sp. strain isolated from milk samples of cows diagnosed with BM, to evaluate their antimicrobial and antibiofilm capacity. The NQo showed great antibacterial effect in the minimum inhibitory concentrations (MIC), minimum bactericidal concentration (MBC) and disk diffusion assays. Using sub lethal concentrations, NQo were tested for inhibition of biofilm formation. The results show that the nanoparticles exhibited biofilm inhibition and were capable of eradicate pre-existing mature biofilm. These findings indicate that the NQo could act as a potential alternative to antibiotic treatment of BM.

Antimicrobial Actions and Applications of Chitosan

“…The mechanisms of action of chitosan against bacteria and fungi have been investigated and reported in many articles [ 72 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 ]. Although the antimicrobial properties of chitosan are highly associated with its structure, physicochemical characteristics, and environmental conditions, in addition to the reactive hydroxyl groups at the C-3 and C-6 positions [ 4 , 37 , 39 , 43 , 90 , 103 , 105 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 ], the mode of action of chitosan against microbes can be classified as extracellular effects, intracellular effects, or both based on the targeting site of the antimicrobial effects [ 36 , 68 ,…”

“…A major antimicrobial effect of chitosan is electrostatic interactions between this cationic molecule and negatively charged cell walls [ 72 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 ]. The pKa values of the amino groups of chitosan are 6.3–6.5, indicating that it is insoluble in alkaline solutions, organic solvents, and water when the pH is higher than 6.5 [ 36 , 68 , 90 , 103 , 107 , 121 ].…”

“…Chitosan nanoparticles are synthesized for different purposes by various methods, such as ionotropic gelation, polyelectrolyte complexation, emulsification solvent diffusion, microemulsion, and reverse micelle formation [ 204 , 205 ]. Moreover, the antimicrobial effects of chitosan-based nanoparticles for drug and drug-free delivery systems have been intensively investigated against bacteria and fungi [ 56 , 57 , 58 , 60 , 63 , 67 , 68 , 106 , 119 , 203 , 206 , 207 , 208 , 209 , 210 , 211 , 212 ]. For example, chitosans of different DDAs and MWs exhibited synergistic activity with sulfamethoxazole against P. aeruginosa [ 213 ].…”

Enhancement of Nano-Biopolymer Antibacterial Activity by Pulsed Electric Fields

“…To demonstrate such a new combination, the antibacterial effects of ChNPs and ELF-PEFs were tested against Gram positive and negative bacteria models as an example of highly infectious wounds. In chronic wounds, Staphylococcus aureus ( S. aureus ) and Pseudomonas aeruginosa ( P. aeruginosa ) colonize about 93.5% and 52.2% of patients with chronic leg ulcers, respectively [ 26 , 27 , 28 , 29 ]. The bacterium P. aeruginosa has the ability to establish itself in biofilms [ 30 , 31 ].…”

A Potential Quorum-Sensing Inhibitor for Bronchiectasis Therapy: Quercetin–Chitosan Nanoparticle Complex Exhibiting Superior Inhibition of Biofilm Formation and Swimming Motility of Pseudomonas aeruginosa to the Native Quercetin

“…The nanoplex was prepared by a simple drug-polyelectrolyte complexation technique that involved only bulk mixing of the ionized drug solution (i.e., QUE) and the oppositely charged polyelectrolytes (PE) (i.e., chitosan, CHI). CHI was selected as the PE for two reasons; first, CHI is a strong polycation hence capable of electrostatic binding with the anionic QUE, and second, CHI is known to possess potent antibacterial activities making it an ideal adjuvant for incorporation into bronchiectasis’ treatment regimen [ 29 , 30 ].…”