Effect of Silver Nanoparticles on Biofilm Formation by Clostridium perfringens Isolated from Poultry and Molecular Typing of Strains by ERIC-PCR

Abstract

The aim of this is study was to evaluate of antibacterial activity of silver nanoparticles (AgNPs) on multidrug resistant (MDR) Cl. perfringens strains isolated from broilers as an alternative to conventional antibiotics and Molecular typing of the C. perfringens strains using ERIC-PCR for assessment of genetic relationship between strains isolated from different organs. A total of 20 isolates of Cl. perfringens were isolated from necrotic enteritis affected poultry located in El-Behera Governorate, Egypt. Antimicrobial resistance pattern was evaluated. ERIC- PCR genotyping of Cl. perfringens   isolates was performed to detect the genetic diversity and the degree of similarity between isolates. Antibacterial activity of AgNPs on MDR strains was evaluated by the minimum inhibitory concentration (MIC) and, minimum bactericidal concentration (MBC). Transmission Electron Microscopy (TEM) imaging was conducted to evaluate the effect of silver nanoparticles on C. perfringens cells. Scanning Electron Microscopy (SEM) imaging used to evaluate the effect of AgNPs on Cl. perfringens biofilm formed on glass cover slide for different periods of incubation with the same concentration of silver nanoparticles. Out of 85 collected samples from broiler chicken farms, 20 isolates of Cl. perfringens type A were isolated from broiler chickens at El-Behera governorate, Egypt. Most of isolates were MDR. ERIC PCR genotyping classified the isolates into 4 clusters (C1-C4).  MBC was 30ug/ml, while MIC was 20ug/ml and the bacterial growth completely inhibited after 24 hours. TEM and SEM images showed that AgNPs has exerted broad-spectrum bactericidal activity against MDR Cl. perfringens isolate and against biofilm.