Cellular injuries upon exposure of Escherichia coli and Lactobacillus rhamnosus to high-intensity ultrasound.
OBJECTIVE
To examine cellular injuries occurring in cells of Escherichia coli (Gram-negative bacteria) and Lactobacillus rhamnosus (Gram-positive bacteria) in response to a high-intensity ultrasound treatment using classical plate count technique and flow cytometry.


RESULTS
According to plate count results, E. coli (D-value 8.3 min) was far more sensitive than L. rhamnosus (D-value 18.1 min) in their response to the ultrasound intensity applied (20 kHz, 17.6 W). The dye precursor carboxyfluorescein diacetate (cFDA) could freely diffuse across the cytoplasmic membrane of intact cells of Gram-positive bacteria L. rhamnosus, resulting in its intracellular enzymatic conversion and emission of green fluorescence. In contrast, the presence of an outer membrane on E. coli, which represents the class of Gram-negative bacteria, apparently disabled the penetration of viability marker cFDA. Ultrasound application on E. coli yielded in an increasing population with disintegrated outer membrane, which allowed penetration of cFDA and its intracellular enzymatic conversion as well as accumulation. In both organisms evaluated only a small population was labelled by propidium iodide upon exposure to ultrasound for up to 20 min. Within the experimental conditions investigated ultrasound did not considerably affect the cytoplasmic membrane, although according to plate count results viability loss occurred.


CONCLUSIONS
The results compiled suggest, that ultrasound induced cell death, which may not be related to membrane damage.


CONCLUSIONS
Limitation on the use of bacteriocins, which are aimed on destabilization of cytoplasmic membrane but inhibited by the outer membrane, could be overcome by ultrasound-assisted physical disruption of the outer membrane.
Ananta
E
E
Department of Food Biotechnology and Food Process Engineering, Berlin University of Technology, Berlin, Germany.
Voigt
D
D
Zenker
M
M
Heinz
V
V
Knorr
D
D
D002462:Cell Membrane / Q000502:physiology; D015169:Colony Count, Microbial / Q000379:methods; D004058:Diffusion; D004926:Escherichia coli / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods; D005452:Fluoresceins / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D005516:Food Microbiology; D006358:Hot Temperature; D007778:Lactobacillus / Q000502:physiology*; D014465:Ultrasonics
D005452:Fluoresceins; D005456:Fluorescent Dyes; C027780:carboxyfluoresceindiacetate