Raman Spectroscopy Applied to Early Detection of Clostridium Infection in Milk

Abstract

Detecting Clostridium in milk presents a significant challenge for the dairy industry given that traditional methods are time-consuming and not specific for these bacteria. Microbiological techniques are expensive and require qualified personnel. Clostridium, in the form of spores, can withstand pasteurization and revert to its vegetative form during cheese aging. These gas-producing bacteria are known for their production of carbon dioxide and hydrogen, causing the formation of slits, cracks, and irregular eyes in hard and semi-hard cheeses. However, gas analysis in the vial headspace of appropriate culture can be exploited to specifically detect Clostridium presence, since the closest competing bacterial Bacilli produces only carbon dioxide. The aim of this paper is to present a Raman-spectroscopy-based instrument for a rapid, inexpensive identification of Clostridium in milk with a limit of detection of 29 spores/L. The proposed measurement procedure is analog to that routinely used, based on the most probable number method. The Raman-based instrument speeds up the detection of a vial's positivity. A test conducted with Clostridium spores demonstrated its effectiveness in almost halving the time needed for the measurement campaign compared to the traditional method.