Near-infrared spectroscopy coupled with chemometrics algorithms for the quantitative determination of the germinability of Clostridium perfringens in four different matrices.

ABSTRACT

Clostridium perfringens (C. perfringens) has the ability to form metabolically-dormant spores that can survive food preservation processes and cause food spoilage and foodborne safety risks upon germination outgrowth. This study was conducted to investigate the effects of different AGFK concentrations (0, 50, 100, 200 mM/mL) on the spore germination of C. perfringens in four matrices, including Tris-HCl, FTG, milk, and chicken soup. C. perfringens spore germinability was investigated using near infrared spectroscopy (NIRS) combined with chemometrics. The spore germination rate (S), the OD600%, and the Ca2+-DPA% were measured using traditional spore germination methods. The results of spore germination assays showed that the optimum germination rate was obtained using 100 mM/L concentrations of AGFK in the FTG medium, and the S, OD600% and Ca2+-DPA% were 98.6%, 59.3% and 95%, respectively. The best prediction models for the S, OD600% and Ca2+-DPA% were obtained using SNV as the preprocessing method for the original spectra, with the competitive adaptive weighted resampling method (CARS) as the characteristic variables related to the selected spore germination methods from NIRS data. The results of the S showed that the optimum model was built by CARS-PLSR (RMSEV = 0.745, Rc = 0.897, RMSEP = 0.769, Rp = 0.883). For the OD600%, interval partial least squares regression (CARS-siPLS) was performed to optimize the models. The calibration yielded acceptable results (RMSEV = 0.218, Rc = 0.879, RMSEP = 0.257, Rp = 0.845). For the Ca2+-DPA%, the optimum model with CARS-siPLS yielded acceptable results (RMSEV = 44.7, Rc = 0.883, RMSEP = 50.2, Rp = 0.872). This indicated that quantitative determinations of the germinability of C. perfringens spores using NIR technology is feasible. A new method based on NIR was provided for rapid, automatic, and non-destructive determination of the germinability of C. perfringens spores.