Abstract
Antibiotic resistance in bacterial pathogens has become a worldwide public health problem in veteri- nary and human medicine. The emergence of methicillin resistance in Staphylococcus sp. has led to complications in treatment of bovine mastitis, wound infections in horses, dogs, pigs etc. and bacterimia in humans ranging from postules, sepsis to even death. To address this problem of antibiotic resistance, several attempts are being made of which, phage therapy offers a promising alternative. Herein, we report a phage based antibacterial nanoplatform hav- ing bactericidal potential against an isolated methicillin resistant skin bacterium, Staphylococcus arlettae as a model organism. The bacteriophages (phages) used in the study were isolated from sewage water via enrichment method against an isolated methicillin resitant skin bacterium, Staphylococcus arlettae (GenBank accession no. KF589201.1). The isolated phages were characterized thereafter for their host specificity and morphology by plaque assay and transmission electron microscopy (TEM) respectively. Intrinsic charge characteristic of phages was utilized for their immobilization on silica NPs monolyer using different conjugation chemistry, through covalent/electrostatic/ phy- sisorption interactions. The efficacy of each immobilization approach was compared using plaque assay. Silica NPs of size ~ 300 nm were synthesized by hydrolysis and condensation of tetraethylorthosilicate in an alkaline envi- ronment, and characterized using scanning electron microscopy (SEM). The particles were dispersed at air/water interface for monolayer formation on polyvinyllidine fluoride (PVDF) membrane via Langmuir-Blodgett (LB) thin film deposition technique at 15 mN/m surface pressure. It was observed that 3-aminopropyltrimethoxysilane (APTMS) treated membranes act as a better bactericidal platform due to proper orientation of phages by means of electrostatic and covalent interaction as compared to phage bound by physical adsorption. Moreoever, high sur- face/volume ratio of NPs further helped in improving bactericidal performance, associated with large loading ca- pacity of phages on film. Thus, the present study can facilitate in design and development of an effective anti-bac- terial compositions and dressings in veterinary medicine and animal agriculture to combat Staphylococcal infections.
Highlights
Staphylococcus sp. is an important gram-positive bacterial pathogen responsible for several infections in animals such as soft tissue abscesses, wound infections, endocarditis, osteomyelitis, bacteremia, mastitis etc. and is a May 2015 | Volume 3 | Special issue 1 | Page 1 cause of great concern worldwide
The emergence of antibiotic resistance in Staphylococcus sp. and ability to persist undetected in herds further apprehends the need for their control measures (DeGraves and Fetrow, 1993; Mann, 2008)
Bacteriophages have emerged as promising alternative to antibiotics and have shown to be one of the highly efficient and effective solutions for controlling Staphylococcal infections as their life cycle is connected exclusively with bacterial cells (Banowski and Simmering, 2011; Donovan, 2007)
Summary
Staphylococcus sp. is an important gram-positive bacterial pathogen responsible for several infections in animals such as soft tissue abscesses, wound infections, endocarditis, osteomyelitis, bacteremia, mastitis etc. and is a May 2015 | Volume 3 | Special issue 1 | Page 1 cause of great concern worldwide. Bacteriophages have emerged as promising alternative to antibiotics and have shown to be one of the highly efficient and effective solutions for controlling Staphylococcal infections as their life cycle is connected exclusively with bacterial cells (Banowski and Simmering, 2011; Donovan, 2007) These are stable macromolecular assemblies, which can maintain their ability to infect for decades, and are relatively insensitive to wide range of temperature, pH and ionic strength (Pearson et al, 2013; Kutter and Sulkvelidze, 2005).The antibacterial acitivity of bacteriophages has been exploited for their applications as sanitizers, disinfectants, food packagings and in detection of bacterial pathogens (Shin et al, 2011; Singh et al, 2009; Yang et al, 2013). The phages were conjugated to silica nanoparticles (NPs) Langmuir-Blodgett (LB) film in an oriented manner using different surface chemistries and their bactericidal effectiveness was investigated in comparison to random immobilisation by physical adsorption method
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