Authors:J. Farkas, É. Andrássy, A. Simon, and L. Mészáros
A recombinant Bacillus subtilis strain containing a plasmid encoding a luxAB fusion, which gave bioluminescence upon addition of an exogenous long-chain aldehyde as substrate for the endogenous luciferase enzyme, was used as test organism. Its populations were treated with 300 MPa for 20 min, or 600 MPa for 20 min at around room temperature, and this treatment is foreseen as a quality-friendly, non-thermal pasteurisation of foods. Besides the estimation of viable cell counts, the extent of pressure-induced germination and post-process development were investigated by phase-contrast microscopy, turbidimetry and luminometry. Increased heat sensitivity of pressurized spore populations was observed both by viable cell counting during a linearly programmed elevation of temperature and a simultaneous differential scanning calorimetry. This was related to pressure-induced germination of spores, although a small fraction remained ungerminated. The luciferase pool built into the spores during their formation seemed to have withstood pressurization. Spore germination was accompanied by the emergence of bioluminescence which also indicated sensitively the characteristic changes of metabolic activity running parallel with the development of untreated cell populations and that of the survivors of the hydrostatic pressure treatments when the cells were incubated in a nutrient broth.
Authors:K. Horváth, Zs. Seregély, É. Andrássy, I. Dalmadi, and J. Farkas
Experimental batches of chilled boneless slices of pork meat have been stored aerobically in sterile Petri dishes and total aerobic plate counts (TAPC) and sensorial observations were made periodically during storage to monitor bacterial growth and apparent deteriorative changes at 4, 8 and 12 °C, respectively. Near infrared spectroscopy (diffuse reflectance) measurement was performed on replicate meat samples in the wavelength range of 1000–1800 nm. Second derivative and multiplicative scatter correction were performed on the spectra as data pre-treatments. Principal component analysis (PCA) and canonical discriminant analysis (CDA) were used for observation of discrimination of the samples due to loss of freshness and onset of bacterial spoilage as a function of the storage time. The percentage of correctly classified samples decreased somewhat by increasing the storage temperature. Partial least squares (PLS) chemometric model was developed to predict and quantify bacterial loads from the scatter corrected 2nd derivative spectra. PLS evaluation (predicted versus measured TAPC values) — when bacterial counts at all sampling days and storage temperatures were taken into account — resulted in a correlation coefficient of 0.977, and a root mean square error of prediction (RMSEP) 0.438 log colony forming units/g. These preliminary results indicate the potential of utilising near infrared diffuse reflectance spectroscopy in combination with multivariate statistical methods to monitor loss of freshness and detect bacterial spoilage of meat samples rapidly before deleterious microbial changes become apparent. However, much larger number of samples should be studied to ascertain properly the prediction power of the spectroscopic method.
Authors:K. Horváth, É. Andrássy, M. Korbász, and J. Farkas
Experimental batches of chilled cutlets of pork have been stored aerobically and surface growth of bacteria was determined by standard aerobic spread-plate counts, and colony counts of pseudomonads on Cetrimid Agar simultaneously with detection times of respective stomached samples in triplicate using a Malthus Microbiological Analyzer at 30 °C, applying general impedance broth and
impedance medium, respectively. To study the effects of storage temperature on growth of spoilage bacteria, numerous samples were kept in successive experiments at 4, 8 or 12 °C. Parameters of the bacterial growth curves were estimated by curve fitting of the colony counts by DMfit programme package of the ComBase softwares, using Baranyi’s dynamic growth model. Linear correlations were found between the respective colony counts and the conductimetric detection times observed in periodic investigations during storage. According to these calibrations, the conductimetric method at 30 °C incubation is able to assess within 8 h whether a sample of pork cutlets contains greater or less than 10
of psychrotrophic spoilage bacteria.
Authors:J. Farkas, É. Andrássy, Z. Formanek, and L. Mészáros
A bioluminescent derivative of Bacillus subtilis containing a plasmid encoding a luxAB fusion under control of a vegetative promoter and gives bioluminescence upon addition of an exogenous long-chain aldehyde has been used as test organism. Its spore populations have been produced and their heat- and radiation survival curves established. Heat-sensitization effect of pre-irradiation of spores was proven not only by colony counting but also with differential scanning calorimetry. Under a linearly programmed temperature increase, the heat destruction of spores surviving 2.5 kGy gamma irradiation resulted in at a few centigrade lower temperature than that of untreated spores. Heat denaturation endotherms in the DSC-thermogram of irradiated spores were shifted to lower temperatures as well. Comparative turbidimetric, luminometric and phase-contrast microscopic studies of untreated, heat-treated and irradiated spore populations showed that the kinetics of germination and the light emission during germination of radiation-inactivated spores were the same as those of untreated spores, revealing that the pre-formed luciferase enzyme packaged into the spores during sporulation remained intact after an irradiation dose causing 90% decrease in number of colony forming spores. Therefore, in contrast to heat-treated spores, the initial bioluminescence reading upon germination of irradiated spores does not reflect the viable count of their population.
Authors:É. Andrássy, J. Farkas, Zs. Seregély, I. Dalmadi, E. Tuboly, and V. Lebovics
Experiments were performed to study changes caused by irradiation or high hydrostatic pressure pasteurization of liquid egg white by differential scanning calorimetry, spectrofluorimetry, electronic nose measurements and NIR-spectrometry. The non-thermal pasteurization treatments were also assessed in relation to loss of carotenoid content, and lipid- and cholesterol oxidation of liquid egg yolk. Unlike radiation pasteurization, high pressure processing caused protein denaturation in egg white, which manifested in changes of its DSC-thermogram and intrinsic tryptophan fluorescence. Electronic nose testing showed changes of the head-space volatile composition of egg albumen, particularly as a function of radiation treatment. Both treatments caused changes in the NIR-spectrometric “fingerprint” of the liquid egg white. Various chemometric analyses of the results of the latter instrumental methods, particularly statistical techniques developed by the group of one of the co-authors of this article, demonstrated the potential for detection and characterization of the applied non-thermal processing techniques on liquid egg white. Irradiation induced more carotenoid degradation and lipid oxidation in liquid egg yolk than pressure processing.
Authors:R. Juhász, K. Horváth, I. Dalmadi, É. Andrássy, A. Salgó, and J. Farkas
Effect of 60Co irradiation on wheat and white pepper grains were investigated in this study using Rapid Visco Analyser (RVA), near infrared reflectance spectroscopy and differential scanning calorimetry. Functional properties of wheat and white pepper were affected by irradiation indicated by a decrease in viscosity values. It was caused by changes of starch structure confirmed by the NIR spectra changes between wavelength 1560–1620 nm, which is the vibration of intermolecular hydrogen bonded OH groups in polysaccharides. The radiation used did not cause significant changes in the thermal properties. RVA proved to be useful for screening radiation induced changes in dry commodities of considerable large starch content on the basis of their rheological behaviour.
Authors:Cs. Mohácsi-Farkas, J. Farkas, L. Mészáros, O. Reichart, and É. Andrássy
Thermal stability of vegetative cells of Listeria monocytogenes, Escherichia coli and Lactobacillus plantarum was studied by counting viable fractions and determining DSC curves of their suspensions. DSC curves in the 5–99°C range showed a series of endothermic transitions between 50 and 60°C, where the heat destruction of cells occurred. Heat denaturation of DNA required a higher temperature than cell killing. Thermal death was strongly influenced by the pH, composition and NaCl content of the suspending buffer. A mathematical model developed by us enabled comparison of DSC peak temperatures and temperatures required for loss of viability.
Authors:J. Csapó, J. Schmidt, Zs. Csapó-Kiss, G. Holló, I. Holló, L. Wágner, É. Cenkvári, É. Varga-Visi, G. Pohn, and G. Andrássy-Baka
In the past years several methods have been developed for the determination of the proportion of the nitrogen-containing substances of microbial origin passed from the rumen into the abomasum or the small intestine. Recently, on examining the D-amino acid content of foodstuffs, particularly milk and milk products, it has been observed that, in addition to D-Ala, D- glutamic acid (D-Glu) and D-aspartic acid (D-Asp) can also be detected in similar quantities, primarily in products which have links with bacterial activity. This gave rise to the idea of examining the diaminopimelic acid (DAPA), D-Glu and D-Asp content of bacteria extracted from the rumen of cattle and that of chyme from the same cattle, in order to determine the type of relation existing among these three components, and to establish whether D-Asp and D-Glu can be used in the estimation of protein of bacterial origin. On determination of the DAPA, D-Asp and D-Glu content by means of amino acid analyser and high performance liquid chromatography of duodenal chyme from five growing bulls and of ruminal bacteria from the same bulls, the following values were established. For chyme (and, in brackets, for ruminal bacteria) r value calculated by means of linear regression was 0.78 (0.76) between DAPA and D-Asp, and 0.70 (0.81) between DAPA and D-Glu. The r values between the crude protein content of ruminal bacteria and the markers examined were found to be the following: DAPA, 0.74; D-Asp, 0.73; D- Glu, 0.61. In the model experiment performed for the re-obtaining of values for protein of bacterial origin the theoretical values were determined on the basis of D-Asp and D-Glu and values approximately 10% higher than the theoretical value on the basis of DAPA. It is therefore recommended that in addition to DAPA these other two amino acids be included among the bacterial protein markers.