Differentiation of four Aspergillus species and one Zygosaccharomyces with two electronic tongues based on different measurement techniques.

Two electronic tongues based on different measurement techniques were applied to the discrimination of four molds and one yeast. Chosen microorganisms were different species of Aspergillus and yeast specie Zygosaccharomyces bailii, which are known as food contaminants. The electronic tongue developed in Linköping University was based on voltammetry. Four working electrodes made of noble metals were used in a standard three-electrode configuration in this case. The St. Petersburg electronic tongue consisted of 27 potentiometric chemical sensors with enhanced cross-sensitivity. Sensors with chalcogenide glass and plasticized PVC membranes were used. Two sets of samples were measured using both electronic tongues. Firstly, broths were measured in which either one of the molds or the yeast grew until late logarithmic phase or border of the stationary phase. Broths inoculated by either one of molds or the yeast was measured at five different times during microorganism growth. Data were evaluated using principal component analysis (PCA), partial least square regression (PLS) and linear discriminant analysis (LDA). It was found that both measurement techniques could differentiate between fungi species. Merged data from both electronic tongues improved differentiation of the samples in selected cases.

Use of an electronic tongue and HPLC with electrochemical detection to differentiate molds in culture media.

A study was conducted to further evaluate an electronic tongue, using high-performance liquid chromatography (HPLC) with electrochemical (EC) and UV detection as a reference method. The electronic tongue consisted of four working electrodes made of different metals and arranged in a standard three-electrode configuration. Pulses of voltage were applied to the metals, and the current responses were sampled and collected in a data matrix. The objectives of the present investigation were to examine the ability of the electronic tongue to distinguish between two mold species growing in three different media, and to obtain support for the hypothesis that the device actually discriminates between different redox-active metabolites produced by the molds. Peak areas in EC and UV HPLC chromatograms were collected in a data matrix. The electronic tongue data and the EC and UV data were then subjected to principal component analysis (PCA). A number of peaks in the HPLC-EC chromatograms indicated that the growth media contained redox-active metabolites. Moreover, PCA of peak areas in EC chromatograms revealed differences between the distribution of redox-active metabolites produced by the two species and between the three culture media. The same pattern was apparent in a PCA score plot of electronic tongue data. The peaks in the UV and EC chromatograms differed, and these were also shown by the PCA score plots.

Use of an electronic tongue to analyze mold growth in liquid media.

The feasibility of employing an electronic tongue to measure the growth of mold in a liquid medium was studied. We used the electronic tongue developed at Linköping University, which is based on pulsed voltammetry and consists of an array of different metal electrodes. Instead of focusing on a single parameter, this device provides information about the condition or quality of a sample or process. Accordingly, the data obtained are complex, and multivariate methods such as principal component analysis (PCA) or projection to latent structures (PLS) are required to extract relevant information. A gas chromatographic technique was developed to measure ergosterol content in mold biomass and was subsequently used as a reference method to investigate the ability of the electronic tongue to measure the growth of mold in liquid media. The result shows that the electronic tongue can monitor mold growth in liquids. In PLS analysis, the electronic tongue signals correlate well with the amount of ergosterol in the mold biomass as well as the microbially induced changes in the pH of the medium.

Metal loading and enzymatic degradation of fungal cell walls and chitin.

The capacity of chitin (from crab shells) and of fungal cell walls from Trichoderma harzianum to accumulate zinc, cadmium and mercury was studied as well as the effects of adsorbed metals on the enzymatic hydrolysis by Novozym 234 of the two substrates. The total adsorbing capacity with respect to these metals was estimated to be at least 10 mmol kg-1 chitin (dry weight) and 50 mmol kg-1 fungal cell walls (dry weight), respectively, at pH 6.1. Enzymatic digestion of fungal cell walls preloaded with mercury and cadmium was significantly reduced, while zinc did not cause any significant inhibition. The effect of metal complexation by chitin on the enzymatic digestion was not as pronounced as for fungal cell walls. This could reflect the fact that chitin sorbed a lower total amount of metals. The inhibitory effect of metals on the enzymatic hydrolysis was caused by the association of the metals with the two substrates and not by the presence of free metals in solution.