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Acoustic Emission Signal of Lactococcus lactis before and after Inhibition with NaN3 and Infection with Bacteriophage c2

DOI: 10.1155/2013/257313

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Abstract:

The detection of acoustic emission (AE) from Lactococcus lactis, ssp lactis is reported in which emission intensities are used to follow and define metabolic activity during growth in nutrient broths. Optical density (OD) data were also acquired during L. lactis growth at 32°C and provided insight into the timing of the AE signals relative to the lag, logarithmic, and stationary growth phases of the bacteria. The inclusion of a metabolic inhibitor, NaN3, into the nutrient broth eliminated bacteria metabolic activity according to the OD data, the absence of which was confirmed using AE data acquisition. The OD and AE data were also acquired before and after the addition of Bacteriophage c2 in L. lactis containing nutrient broths during the early or middle logarithmic phase; c2 phage m.o.i. (Multiplicity of infection) was varied to help differentiate whether the detected AE was from bacteria cells during lysis or from the c2 phage during genome injection into the cells. It is proposed that AE measurements using piezoelectric sensors are sensitive enough to detect bacteria at the amount near ?cfu/mL, to provide real time data on bacteria metabolic activity and to dynamically monitor phage infection of cells. 1. Introduction Periodic, vibrational motion of Saccharomyces cerevisiae yeast cell walls was observed using atomic force microscopy (AFM) [1, 2]. The wall vibrations which were temperature dependent, had frequencies between 1.0–1.6?kHz and ceased after the addition of a metabolic inhibitor. The periodic vibrations were ascribed to forced, concerted cell wall motions caused by cellular metabolism and molecular motors such as kinesin, dynein, and myosin rather than to natural resonant cell wall oscillations. The cell wall amplitudes were 3?nm with forces ~10?nN; for human foreskin fibroblasts the cell wall motion from actin-myosin activity produced forces between 20–100?pN. Hence, the range of work ((force) × (amplitude)) associated with cell wall motion in eukaryotes can be estimated to be between ~ ?J-to- ?J (2000?aJ-to-30?aJ). These values are well above the < ?aJ sensitivities known for commercially-acquired piezoelectric-based sensors. The routine application of AFM techniques to study whether periodic motion of bacteria cell walls occurs during metabolism may be considerably more difficult than of yeast cell walls because bacteria have less rigid walls and are much smaller in size. Nevertheless, bacterial wall vibration as caused by molecular motors has been theoretically modeled to produce vibrational motion with frequencies up to 10?kHz [3]. To

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