Classification of Swiss cheese starter and adjunct cultures using Fourier transform infrared microspectroscopy

Abstract

The acceptability of Swiss cheese largely depends on the flavor profile, and strain variations of cheese cultures will affect the final quality. Conventional biochemical methods to identify the cultures at the strain level are time-consuming and expensive, and require skilled labor. Our objective was to develop rapid classification methods of starter cultures at the strain level by using a combination of hydrophobic grid membrane filters and Fourier transform infrared (FTIR) spectroscopy. Forty-four pulsed-field gel electrophoresis–verified strains of starter and nonstarter cultures including Streptococcus thermophilus, Propionibacterium freudenreichii, and Lactobacillus spp. were analyzed. The strains were grown on their respective agar media, transferred to broth media, and incubated. Then, cultures were centrifuged and the pellets were resuspended in saline solution (10μL). Aliquots (2μL) of the suspended bacterial solution were placed onto a grid of the hydrophobic grid membrane filters, having 6 grids per each strain analyzed. The dried filters were read by FTIR microspectroscopy fitted with an attenuated total reflectance probe. Collected spectra were statistically analyzed by a soft independent modeling of class analogy (SIMCA) for pattern recognition. Classification models were developed for Streptococcus thermophilus, Propionibacterium freudenreichii, and Lactobacillus spp. strains. The models showed major discrimination in the spectral region from 1,200 to 900cm−1 associated with signals from phosphate-containing compounds and various polysaccharides in the cell wall. The developed method allowed for rapid classification of several Swiss cheese starter and nonstarter cultures at the strain level. This information provides a detailed overview of microbiological status, which would enable corrective measures to be taken early in the cheese making process, limiting production of inferior quality cheese and minimizing defects. This method could be an effective tool to identify and monitor activity of cheese and other dairy starter cultures.