Construction and characterization of bioluminescentReading outbreak and non-outbreak strains.

Abstract

UNLABELLED: enterica serotype Reading has recently been identified as a significant foodborne pathogen from contaminated poultry products. There is a critical need for close monitoring of this newly emerged pathogen. This study developed bioluminescent strains ofReading for real-time pathogen tracking using bioluminescence imaging. Two strains of. Reading were used: an outbreak strain and a non-outbreak strain. The chloramphenicol acetyltransferase gene was cloned into the plasmid pBS-GFP, which carriesoperon and an ampicillin resistance gene. The newly constructed plasmid was then transformed into the outbreak and non-outbreak strains of. Reading by electroporation. The resulting colonies were confirmed by visualizing bioluminescence using animaging system and by testing their resistance to chloramphenicol. These strains demonstrated a high bioluminescence level (10-10Photons/s/cm/sr) and were tested for growth and plasmid stability by daily subculturing in Luria-Bertani medium with and without antibiotics. The plasmid remained stable for 8 days under non-selective conditions, and growth rates were comparable to non-bioluminescent parent strains in antibiotic-free conditions. However, growth was notably different in the presence of chloramphenicol, indicating successful plasmid retention and function. This study successfully created stable bioluminescent. Reading strains, marking a significant step forward in monitoring and potentially reducing the spread of this emergent foodborne pathogen in the poultry industry. IMPORTANCE: serotype Reading has recently become a significant foodborne pathogen linked to poultry products. To enhance pathogen monitoring, this study developed bioluminescent strains ofReading by inserting the chloramphenicol acetyltransferase gene into a plasmid containing a bioluminescence gene cluster. These modified strains were transformed into outbreak and non-outbreak bacterial strains via electroporation. The bioluminescent strains demonstrated stable plasmid retention and high bioluminescence levels. They also showed growth comparable to their parent strains, even in the absence of antibiotics. These bioluminescent strains could potentially facilitate real-time monitoring and control ofReading in poultry industries.