Rapid serotype-independent differential detection of biofilm-positive and biofilm-negative Salmonella using Fourier transform infrared biotyping

Abstract

Foodborne illnesses caused by Salmonella represent a global one health challenge, with biofilm-forming strains exhibiting enhanced public health risks due to their ability to persist due to resistance to antimicrobial agents, disinfectants, and environmental stresses. While food-safety and public health investigation primarily focus on Salmonella identification and source tracing, they often overlook the biofilm-forming capacity of isolates, limiting their predictive value for risks posed by biofilm producing Salmonella. This study assessed fourier transform infrared (FTIR) biotyping for rapid serotype-independent differentiatial detection of biofilm-positive (BFP) from biofilm-negative (BFN) Salmonella. A total of 270 Salmonella strains representing 12 common serotypes were classified using three conventional biofilm assays (congo red and coomassie brilliant blue agar test, calcofluor test, and tube test) into true BFP (n = 80), true BFN (n = 64), and uncertain (n = 59) biofilm producers. Biofilm production for each group was also assessed with a microtiter plate assay. FTIR biotyping was applied to a subset of 115 strains (61 BFP, 54 BFN). Using spectral windows of 1180–1050 cm⁻¹ and 1400–1200 cm⁻¹, FTIR biotyping accurately differentiated BFP from BFN strains with 93.4 % sensitivity, 83.3 % specificity, and 88.6 % overall accuracy. FTIR biotyping differentiated 59 strains with uncertain biofilm status into BFN (n = 45) and BFP (n = 14). FTIR biotyping provides a rapid, sensitive and specific method for detection of biofilm-forming Salmonella strains. Incorporating FTIR biotyping for biofilm detection in current Salmonella surveillance and source-tracing protocols can enhance food safety risk assessments and improve Salmonella outbreak prevention.