FLUORESCENT MICROSCOPY AND GAS CHROMATOGRAPHY TO ASSESS THE VIABILITY AND METABOLIC ACTIVITY OF PROPIONIBACTERIUM SP. STRAINS
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Publication date: 2010-12-31
Pol. J. Food Nutr. Sci. 2010;60(4):363-368
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ABSTRACT
The aim of the study was to assess usefulness of the LIVE/DEAD fluorescent staining method and gas chromatography to monitor the viability and metabolic activity of Propionibacterium strains in long-term cultures in milk. The effect of 4% NaCl addition and a temperature of 10oC on the growth of Propionibacterium freudenreichii ssp. freudenreichii 111, 109C, 108 strains was studied for 28 days. Bacterial cells were assessed in cultures by microscopic and plate counting methods in regular intervals. The cultures were additionally determined for the content of volatile fatty acids: C2 to C7. The total cell counts of all strains in cultures assessed by the microscopic method were noticed to be 1 to 5 logarithmic cycles higher in comparison to those determined with the plate counting method. In following days and weeks of culture, increasing discrepancies were observed between the results obtained using microscopic and plate methods. Both methods revealed similar trends in the viability of strains under control conditions and a little impact of NaCl addition on cell growth and decrease. The cultures run at a temperature of 10oC exhibited different course of growth and decline of the number of monitored populations depending on strain and method applied. Individual strains possessed different acid formation activity. From the beginning of incubation, the highest concentrations were reported for propionic and acetic acids, whereas the other acids in number from 4 to 6 appeared subsequently. The temperature of 10oC inhibited acids formation by all strains, whereas 4% addition of NaCl stimulated the acid-forming activity and during incubation under those conditions the contents of volatile acids were recorded to be the highest.