OWLS APPLICATION NOTES NO-010

EC-OWLS FOR STUDYING THE EFFECT OF DIFFERENT STRESS

FACTORS ON LACTIC ACID BACTERIA
Abstract
Lactic acid bacteria are part of the normal human microflora and play an important role
in the fermentation of different food products, such as milk, cheese or meats. During the
fermentation process lactobacilli are confronted with many inhibitor factors, such as high
salt content, temperature changes, organic acids. It is of great importance to study the
effect of different stress factors on the microbial status of lactic acid bacteria, their
growth, viability and survival.

Lactobacillus plantarum 2142 (Culture Collection of Sensor response of different rate of living and damaged
the Institute of Dairy Microbiology, University of cells
P e r u g i a ) wa s u s e d . F r o z e n c u l t u r e o f t h e l a c t o b a c i l l i D i f f e r e n t r a t i o o f l i v i n g a n d h e a t t r e a t e d c e l l s we r e
was transferred in deMan Rogosa Sharpe (MRS) m i xe d a n d m e a s u r e d . U s i n g t h e s a m e o v e r a l l c e l l
broth (Merck, Darmstadt) using 1% inocula and number, the rate of living cells could be determined.
i n c u b a t e d a t 3 0 °C f o r 2 4 h . B e f o r e e x p e r i m e n t s i t
was subcultured at least two times in sterile MRS
broth.
Electrochemical Optical W aveguide Lightmode
Spectroscopy (EC-OW LS) com bines evanescent-
field optical sensing with electrochemical control of
surface adsorption processes. A layer of indium tin
o xi d e ( I T O ) s e r v e s a s a c o n d u c t i v e e l e c t r o d e f o r
electrochemical sensing and together with the other
glass-type layers of the sensor functioned as a high
refractive index waveguide for optical sensing.
Mass signal of immobilized cells as function of
polarizing potential
Effect of acetic acid treatment on the lactobacillus cells

500 5 M acetic acid

450 1M acetic acid
500 mM acetic acid
400
Mass (arbitrary unit)…..

100 mM acetic acid

350 20 mM acetic acid
300 0 acetic acid
250
200
150
100
50
0
50 60 70 80 90 100
Time (min)
The effect of polarization potential on the number of
immobilized cells, measured as the sensor
response, was studied. The highest response was Effect of hydrogen peroxide treatment on the
o b t a i n e d w h e n 0 . 8 - 1 . 0 V p o t e n t i a l wa s u s e d . A s a lactobacillus cells
result of polarizing potential, the bacterial cells
were adsorbed in native form on the surface of the
sensor.
Calibration curve of lactobacillus cell concentration

References:
Adányi, N., Németh E.; Halász A.; Szendrı I.;
Váradi M.(2006): Application of electrochemical
optical waveguide lightmode spectroscopy
(EC-OW LS) for studyi ng the effect of different
The calibration curve of the bacteria count was stress factors on lactic acid bacteria. Anal Chim
obtained from 102 –108 CFU/mL and by using the
Acta 573-574. 41-47.
average of three replicate measurements. According
to the calibration curve, a typical first-order kinetic
profile in the semi-logarithmic scale can be
observed.

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