Role of fatty acid salts as anti Acanthamoeba agents for disinfecting contact lens.

　Acanthamoeba is found in seawater, fresh water, and soil and is an opportunistic pathogen that causes a potentially blinding corneal infection known as Acanthamoeba keratitis. The anti-amoeba activity of 9 fatty acid salts (potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K), and linolenate (C18:3K)) was tested on Acanthamoeba castellanii ATCC 30010 (trophozoites and cysts). Fatty acid salts (350 mM and pH 10.5) were prepared by mixing fatty acids with the appropriate amount of KOH. C8K, C10K, and C12K showed growth reduction of 4 log-units (99.99% suppression) in A. castellanii upon 180 min incubation at 175 mM, whereas the pH-adjusted control solution showed no effect. After the amoeba suspension was mixed with C10K or C12K, cell membrane destruction was observed. The minimum inhibitory concentration of C10K and C12K was also determined to be 2.7 mM. Confirmation tests were conducted using contact lenses to evaluate the effectiveness of C10K and C12K as multi-purpose solutions. Experiments using increasing concentrations showed reduced numbers of living cells in C10K (5.5 mM, 10.9 mM) and in C12K (5.5 mM, 10.9 mM). These results demonstrate the inhibitory activity of C10K and C12K against A. castellanii and indicate their potential as anti-amoeba agents.

Antibacterial Effect of Fatty Acid Salts on Oral Bacteria.

Fatty acid salts are a type of surfactant known to have potent antibacterial activity. We therefore examined the antibacterial activities of fatty acid salts against Streptococcus mutans. Potassium caprylate (C10K), potassium laurate (C12K), potassium myristate (C14K), potassium oleate （C18:1K), potassium linoleate (C18:2K), and potassium linolenate (C18:3K), used at a concentration of 175 mM, resulted in a 7 log-unit reduction of S. mutans after a 10-min incubation. The minimum inhibitory concentration (MIC) of C18:2K and C18:3K was 5.5 mM. C12K also demonstrated high antibacterial activity (MIC of 21.9 mM). These results indicate that C12K, C18:2K, and C18:3K have high antibacterial activity against S. mutans, and possess great potential as antibacterial agents.

Influence of cations and anions on the induction of cell density-independent luminescence in Photorhabdus luminescens.

Bioluminescence is emitted by various living organisms, including bacteria. While the induction mechanism in marine luminescent bacteria, such as Vibrio fischeri and V. harveyi, has been well characterized, this mechanism has not been studied in detail in the non-marine luminescent bacterium Photorhabdus luminescens. Therefore, we investigated the effect of cations and anions on the induction of luminescence by P. luminescens. Cultivation of cells in an inorganic salts solution (ISS) containing KCl, CaCl2 , MgCl2 , NaHCO3 , and MgSO4 resulted in a rapid increase in luminescence intensity. Moreover, the induction of luminescence in the ISS medium was not dependent on cell density, since cell densities remained unchanged during 48 h. Furthermore, we found that compounds containing K(+) , Mg(2+) , and HCO3(-) were necessary to induce cell density-independent luminescence. The intensity of luminescence per cell cultured in medium containing KCl, MgCl2 , and NaHCO3 was approximately 100-fold higher than that cultured in NB. In contrast, when cells actively grew in normal growth condition, the intensity of luminescence per cell was not increased even in the presence of K(+) , Mg(2+) , and HCO3(-) . Thus, these results suggest that the luminescence of P. luminescens is regulated by 2 independent cell density-dependent and -independent mechanisms.

Selenium cannot substitute for sulfur in cell density-independent bioluminescence in Vibrio fischeri.

It has been proposed that selenium, an element chemically similar to sulfur, can participate in some of the same biological pathways as sulfur, although only a few studies have been confirmed this. In this study, we investigated the relationship between selenium and sulfur-dependent luminescence in Vibrio fischeri. The luminescence of V. fischeri was induced by the addition of sulfur-containing compounds such as Na2SO4 and L-cystine, and their luminescence was suppressed, in a dose-dependent manner, by the addition of the selenium-containing compounds Na2SeO4 and L-selenocystine. Since the viability of V. fischeri was not affected by the addition of low concentration of selenium-containing compounds, the decrease in luminescence intensity cannot be explained by cell death. Kinetic analysis performed using Lineweaver-Burk plots demonstrate that Na2SeO4 and L-selenocystine act as competitive suppressors in inorganic sulfur (Na2SeO4)-dependent luminescence. In contrast, these selenium-containing compounds act as uncompetitive suppressors in organic sulfur (L-cystine)-dependent luminescence.

Requirements for sulfur in cell density-independent induction of luminescence in Vibrio fischeri under nutrient-starved conditions.

Despite the universal requirement for sulfur in living organisms, it is not known whether the luminescence of Vibrio fischeri is sulfur-dependent and how sulfur affects the intensity of its luminescence. In this study, we investigated the requirement for sulfur in V. fischeri luminescence under nutrient-starved conditions. Full induction of V. fischeri luminescence required MgSO(4); in artificial seawater cultures that lacked sufficient MgSO(4), its luminescence was not fully induced. This induction of luminescence was not dependent on autoinduction because the cell density of V. fischeri did not reach the critical threshold concentration. In addition to MgSO(4), this cell density-independent luminescence was induced or maintained by nontoxic concentrations of l-cysteine, sulfate, sulfite, and thiosulfate. Moreover, the addition of N -3-oxo-hexanoyl homoserine lactone and N -octanoyl homoserine lactone, which are known autoinducers in V. fischeri, did not induce luminescence under these conditions. This result suggested that the underlying mechanism of luminescence may be different from the known autoinduction mechanism.

Interactions between bicarbonate, potassium, and magnesium, and sulfur-dependent induction of luminescence in Vibrio fischeri.

In spite of its central importance in research efforts, the relationship between seawater compounds and bacterial luminescence has not previously been investigated in detail. Thus, in this study, we investigated the effect of cations (Na(+) , K(+) , NH(4) (+) , Mg(2+) , and Ca(2+) ) and anions (Cl(-) , HCO(3) (-) , CO(3) (2-) , and NO(3) (-) ) on the induction of both inorganic (sulfate, sulfite, and thiosulfate) and organic (L-cysteine and L-cystine) sulfur-dependent luminescence in Vibrio fischeri. We found that HCO(3) (-) (bicarbonate) and CO(3) (2-) (carbonate), in the form of various compounds, had a stimulatory effect on sulfur-dependent luminescence. The luminescence induced by bicarbonate was further promoted by the addition of magnesium. Potassium also increased sulfur-dependent luminescence when sulfate or thiosulfate was supplied as the sole sulfur source, but not when sulfite, L-cysteine, or L-cystine was supplied. The positive effect of potassium was accelerated by the addition of magnesium and/or calcium. Furthermore, the additional supply of magnesium improved the induction of sulfite- or L-cysteine-dependent luminescence, but not the l-cystine-dependent type. These results suggest that sulfur-dependent luminescence of V. fischeri under nutrient-starved conditions is mainly controlled by bicarbonate, carbonate, and potassium. In addition, our results indicate that an additional supply of magnesium is effective for increasing V. fischeri luminescence.

Effects of magnesium sulfate on the luminescence of Vibrio fischeri under nutrient-starved conditions.

In this study, we investigated the relationship between MgSO(4) and luminescence in Vibrio fischeri under nutrient-starved conditions. When V. fischeri was cultured in an artificial seawater medium, the luminescence intensity was low relative to that observed under normal growth conditions. It decreased during the initial 14 h, and then increased slightly at 24 h. This regulation of luminescence was not dependent on the quorum-sensing mechanism, because the cell densities had not reached a critical threshold concentration. Under MgSO(4)-starved conditions, luminescence was not fully induced at 14 h, and decreased at 24 h. In contrast, induction of luminescence occurred under MgSO(4)-supplemented conditions, but MgSO(4) alone was insufficient to induce luminescence, and required NaHCO(3) or KCl. These results suggest that the luminescence of V. fischeri is controlled by an exogenous sulfur source under nutrient-starved conditions. In addition, they indicate that the induction of sulfur-dependent luminescence is regulated by the NaHCO(3) or KCl concentration.