Bacterial cytoplasmic membrane permeability assay using ion-selective electrodes

Ohmizo, Chie; Yata, Masaya; Katsu, Takashi

doi:10.1016/j.mimet.2004.06.008

Cited by 39 publications

(35 citation statements)

References 22 publications

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“…Moreover, it has been evidenced, using flow cytometry experiments, that the mode of action mediated by bacterial cell permeabilization is similar in both Gram-positive and negative bacteria [17]. The ability to maintain the membrane potential and pH gradient is necessary for cell survival, and a decrease in these parameters is indicative of significant damage to the cell membrane [18].…”

Section: Cytotoxicity Activitymentioning

confidence: 99%

Plant Essential Oil: An Alternative to Emerging Multidrug Resistant Pathogenso

2017

JMEN

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Section: Cytotoxicity Activitymentioning

confidence: 99%

Plant Essential Oil: An Alternative to Emerging Multidrug Resistant Pathogenso

2017

JMEN

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“…perfringens CCM 4435 T was measured by means of the K + ion-selective electrode 20-19+ and AgCl reference electrode 10-251 (2Theta, Ceský Tesin, Czech Republic), connected to a pH/mV meter PHI 04 (Labio, Prague, Czech Republic). A procedure of Ohmizo et al (2004) with a few modifications was used. Bacterial cells were harvested in the exponential phase of growth by centrifugation at 3 000 g for 20 min (centrifuge K-23, Janetzki, Germany).…”

Section: E Colimentioning

confidence: 99%

Susceptibility of Escherichia coli, Salmonella sp. and Clostridium perfringensto organic acids and monolaurin

Skřivanová¹,

Marounek²,

Benda³

et al. 2006

Vet. Med.

176

128

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The antimicrobial activity of fatty acids, monolaurin, citric, succinic, fumaric, malic and lactic acid was determined in cultures of two strains of Escherichia coli, three strains of Salmonella sp. and two strains of Clostridium perfringens. Antimicrobial activity was expressed as minimum inhibitory concentration (MIC) that prevented growth and glucose utilization in treated cultures. Caprylic acid was the only acid inhibiting glucose utilization in all cultures. Its MIC varied from 1 to 3 mg/ml. Strains CCM 3954 and CCM 4225 of E. coli were inhibited also by capric acid at 5 mg/ml. Strains CCM 4435T and CNCTC 5459 of Cl. perfringens were inhibited by medium-chain fatty acids (C8 to C14), oleic acid and one strain also by linoleic acid. The minimum MICs were those of lauric and myristic acid (between 0.1 and 0.2 mg/ml). Growth of Cl. perfringens, but not other bacteria, was inhibited also by monoglyceride of lauric acid (MIC = 3 mg/ml), and by citric acid (MIC = 4 mg/ml). Inhibitory effects of other acids were not observed at 5 mg/ml. Caprylic and lauric acid did not influence the K+ permeability of the cytoplasmic membrane in cells of E. coli CCM 4225 and Cl. perfringens CCM 4435T, respectively. In cultures of both strains of E. coli treated with caprylic acid at 5 mg/ml, and in those of Cl. perfringens CCM 4435T treated with lauric acid at 1 mg/ml, or with its monoglyceride at 5 mg/ml, the transmission electron microscopy revealed damage of cytoplasmatic structures. In cells of Cl. perfringens the separation of inner and outer membranes was apparent, the integrity of the outer membrane, however, was maintained. It can be concluded that medium-chain fatty acids are more efficient antimicrobials than other, more polar organic acids tested.

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“…[1][2][3][4] We were interested in monitoring the time-dependent process of this damage, using electrochemical sensors, such as ion-selective and oxygen electrodes. Although these sensors are widely used to analyze the actions of biologically-active substances in living cells, [5][6][7][8] including photodynamic action, 9 no study has reported the in situ monitoring of the photodynamic inactivation of bacteria under photo-irradiation.…”

mentioning

confidence: 99%

“…Cells were grown at 37 C in a medium containing 1.5% polypeptone, 0.5% bovine extract, 0.5% NaCl and 0.5% K2HPO4. 5 The cells were harvested in the exponential phase of growth, washed twice with buffer (150 mM NaCl and 10 mM NaH2PO4/Na2HPO4 (pH 7.2)) and suspended in this buffer at 4 × 10 10 CFU ml -1 . Figure 1 shows the experimental setup for measuring bacterial respiration.…”

mentioning

confidence: 99%

“…All of the measurements were performed with stirring in a thermostat adjusted to 37 C. The K + and TPP + electrodes were constructed as reported previously. 5,6,14 Melittin (final concentration, 10 μM) was used to disrupt the cytoplasmic membrane of S. aureus cells in order to determine both the 100% level of the K + efflux from bacteria and the 0% level of the membrane potential (the complete dissipation of membrane potential). 5,15 The maximal uptake of TPP + , observed just prior to photo-irradiation, was defined as 100% membrane potential.…”

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confidence: 99%

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