Sorbitol Minimizes Calcium Carbonate Scale Generation While Maintaining the Disinfection Effect of Heated Scallop-Shell Powder for Fresh Produce.

Scallop shells subjected to heat treatment exhibit antimicrobial activity, and heated scallop-shell powder (HSSP) has recently been reported to be effective for disinfecting food. However, because the main component of these shells is calcium oxide, there is a problem that scales of calcium carbonate (CaCO3) become established on the surface of equipment used for food processing. In this study, we thus investigated whether the addition of sugar to HSSP slurry suppressed CaCO3 scale generation and whether the sugar-supplemented HSSP could be applied to the disinfection and preservation of fresh lettuce. The results showed that glucose, sucrose, and sorbitol could suppress the scale generation in HSSP slurry. However, glucose and sucrose decreased the antibacterial activity of HSSP. Since the addition of sorbitol did not affect the antibacterial activity of HSSP slurry, it was used for subsequent experiments because of its low bioavailability. Sorbitol effectively suppressed scale formation by dissolving it before the addition of HSSP. The disinfection and preservative effects of sorbitol-supplemented HSSP ( S-HSSP) treatment on lettuce did not decrease compared with those upon HSSP treatment and were almost equal to or higher than those of sodium hypochlorite treatment at 200 mg/l. The addition of sorbitol solved the major problem of scale generation by HSSP containing CaO, which contributes to expansion of usage of heated shell powder, such as HSSP, in food processing.

Antibacterial Properties of Silicone Membranes after a Simple Two-Step Immersion Process in Iodine and Silver Nitrate Solutions.

　Silicone is widely used in packing materials, medical equipment, and separation membranes. Since microbial cells easily adhere to the surface of silicone materials and form biofilms, techniques for incorporating antimicrobial activity into silicone materials are in high demand. This study describes the preparation of silver (Ag)/silicone composite membranes through a simple two-step immersion process, utilizing an iodine solution followed by a silver nitrate solution at room temperature. Scanning electron microscopy (SEM) observations revealed that particles with sizes of several nanometers to several tens of nanometers were present on the silicone membrane surface; these particles were identified as silver iodide using energy-dispersive X-ray spectroscopy (EDS) . The Ag/silicone membrane possessed excellent antibacterial efficacy against Escherichia coli and Staphylococcus aureus, and the antibacterial efficacy (R) against both types of bacteria was R > 4, even after stomacher treatment or acidic treatment of pH 2-6 for 24 h. The mechanical strength of the silicone membrane was also maintained after antibacterial treatment, with Young's modulus values of 7.9±1.2 MPa and 8.3±1.5 MPa for the untreated membrane and Ag/silicone membrane, respectively (p > 0.05) . In addition, the reduction in permeation performance of the Ag/silicone membrane was only 20%, despite the antibacterial treatment on the membrane surface. This antibacterial treatment method of silicone membranes can be conducted at room temperature (25℃) without special equipment, and may be applied to other types of silicone materials.

Changes in aquatic toxicity of potassium dichromate as a function of water quality parameters.

Potassium dichromate (K2Cr2O7) is used as a general reference toxicant in aquatic toxicity testing, but relatively little is known regarding the effects of water quality parameters on K2Cr2O7 toxicity to Daphnia magna. The acute toxicity of K2Cr2O7 to D. magna was comparatively examined in one very hard (M4 medium for D. magna assay), four hard, one moderately hard and one soft dilution water samples. The 48-h EC50 (50% effective concentration) of K2Cr2O7 to D. magna was reproducible (coefficient of variation [CV]: 13%) in tests using the same dilution water sample, but reproducibility was poor (CV: 62%) in tests using seven different dilution water samples. The observed 48-h EC50 value increased with increasing water hardness (28-250 mg CaCO3/L) and Na+ concentration (4.3-19.7 mg Na/L). The effect of Ca2+ and Mg2+ on K2Cr2O7 toxicity was equivalent in terms of molar concentration. The 48-h EC50 for K2Cr2O7 was determined according to OECD TG 202 by six contract laboratories using M4 medium and were shown to be reproducible (CV: 15%), indicating that the toxicity level can be determined with high accuracy if holding and dilution water samples are standardized. Multiple regression analysis revealed that the 48-h EC50 was strongly correlated (r2 = 0.927) with the Ca2+, Mg2+, Na+ concentration, and alkalinity of the dilution water samples. Detailed monitoring of water quality characteristics thus facilitates intra- and inter-laboratory comparisons of toxicity data and enables predictions of changes in the susceptibility of test animals.

Predicting changes in aquatic toxicity of chemicals resulting from solvent or dispersant use as vehicle.

The influence of two vehicles (N,N-dimethylformamide [DMF] as solvent and polyoxyethylene hydrogenated castor oil [HCO-40] as a dispersant) on the acute toxicity of eight hydrophobic chemicals with a non-specific mode of action to Daphnia magna was investigated according to the OECD Guidelines for the Testing of Chemicals, No. 202. An increased 48-h EC50 value for D. magna or reduced toxicity resulting from the addition of HCO-40 to the test medium was observed for five of the eight chemicals examined. Each of eight chemicals was dissolved in water at a concentration of either 10 mg/L or 1.0 mg/L, with or without DMF or HCO-40. Silicone film as a model of a biological membrane was then immersed in each solution, and the concentration of each chemical in the water was monitored until equilibrium was reached for each test substance, after which the adsorbed amount of each chemical was determined. The amounts of p-pentylphenol and four other substances with log Pow (1-octanol/water partition coefficient) values greater than 3.4 adsorbed onto the silicone film decreased with increasing concentrations of HCO-40. However, 3-chloro-4-fluoronitrobenzene and two other substances with log Pow values less than 2.6 demonstrated no changes in adsorption with either increasing HCO-40 concentration or the addition of DMF. The reduced adsorption in the presence of a vehicle on the silicone film correlated closely with changes in toxicity. These results indicate that the methodology developed in this study enables the prediction of changes in toxicity resulting from the addition of vehicles to a test system.

Antimicrobial Characteristics of Heated Eggshell Powder.

Eggshells have high bioavailability and can be used as a source of calcium. The main component is CaCO3, which, when heated, is converted to CaO. Seashells are also mainly composed of CaCO3 and were previously found to exhibit antimicrobial activity after being heated. In this study, heated eggshell powder (HESP) was found to have antimicrobial activity against bacterial vegetative cells, fungi and bacterial spores. Parameters, such as the minimum inhibitory concentration, were determined with kinetic analysis using an indirect conductimetric assay. Moreover, HESP was able to kill the Bacillus subtilis spores. There were no significant differences in the activity between HESP, heated scallop-shell powder and pure CaO. The MIC values for HESP against bacteria and fungi were 0.29-0.43 and 1.3-1.5 mg/mL, respectively. Against B. subtilis spores, a reduction of two orders of magnitude of viability was confirmed following 20 min of treatment at 10 mg/mL at 60 ℃. The active oxygen generated from the HESP slurry was examined with chemiluminescence. The intensity of this increased with increasing concentrations of the HESP slurry. This suggests that HESP could be used as a natural antimicrobial agent. Although a high pH is the main contributor to this antimicrobial activity, active oxygen species generated from HESP are likely to be the main antimicrobial agents..

Heated scallop-shell powder treatment for deactivation and removal of Listeria sp. biofilm formed at a low temperature.

The ability of heated scallop-shell powder (HSSP) to work against Listeria sp. biofilm formed at a low temperature was investigated. A biofilm of L. innocua ATCC 33090 was grown on a glass plate at 15˚C for 15 days, then immersed in HSSP slurry. Following treatment, the disinfection ability of the HSSP against the biofilm was non-destructively quantified by conductimetric assay. The biofilm grown at 15˚C was less sensitive than that grown at 37˚C to HSSP treatment and alkaline treatment. The biofilm grown at 15˚C was completely deactivated by 30 min of HSSP treatment (10 mg/mL, pH 12.5). In contrast, after 30 min treatment with alkaline solution at pH 12.5 or sodium hypochlorite (100 ppm), the activity was reduced by only one order of magnitude. The disinfection efficacy of HSSP (10 mg/mL) against L. innocua is similar to or higher than that of sodium hypochlorite (200 ppm). Fluorescence microscopy validated the results of the conductimetric assay. Therefore, HSSP treatment is a potentially powerful alternative control agent against Listeria sp. biofilms that present hazards in the food industry.

Sporicidal characteristics of heated dolomite powder against Bacillus subtilis spores.

Dolomite is a double salt composed of calcium carbonate (CaCO3) and magnesium carbonate (MgCO3). The heat treatment of CaCO3 and MgCO3 respectively generates calcium oxide (CaO) and magnesium oxide (MgO), which have antimicrobial activity. In this study, heated dolomite powder (HDP) slurry was investigated for its sporicidal activity against Bacillus subtilis ATCC 6633 spores. The B. subtilis spores used in this study were not affected by acidic (pH 1) or alkaline (pH 13) conditions, indicating that they were highly resistant. However, dolomite powder heated to 1000℃ for 1 h could kill B. subtilis spores, even at pH 12.7. Sporicidal activity was only apparent when the dolomite powder was heated to 800℃ or higher, and sporicidal activity increased with increases in the heating temperature. This temperature corresponded to that of the generation of CaO. We determined that MgO did not contribute to the sporicidal activity of HDP. To elucidate the sporicidal mechanism of the HDP against B. subtilis spores, the generation of active oxygen from HDP slurry was examined by chemiluminescence analysis. The generation of active oxygen increased when the HDP slurry concentration rose. The results suggested that, in addition to its alkalinity, the active oxygen species generated from HDP were associated with sporicidal activity.

Antibacterial characteristics of heated scallop-shell nano-particles.

Heated scallop-shell (HSS) nano-particles, prepared using a wet grinding mill, and microparticles were examined for their antibacterial activity against vegetative bacterial cells and spores. The median diameters of the nano-particles and micro-particles were approximately 20 nm and 30 µm, respectively. The antibacterial activity of HSS against Escherichia coli increased with an increase in concentration, regardless of particle size; however, the antibacterial activity of the nano-particles was much higher than that of micro-particles. The sporicidal activity of the nano-particles was also much higher than that of micro-particles, with HSS nano-particles able to kill Bacillus subtilis spores. A reduction of more than three orders of magnitude for B. subtilis spores was confirmed following a 30 min treatment at 5 mg/ml and 60℃, showing that the combination of HSS nano-particle treatment with mild heating was particularly effective for controlling bacterial spores.

Bacterial degradation and reduction in the estrogen activity of 4-nonylphenol.

Bacteria capable of degrading 4-nonylphenol (NP) were isolated and identified, and their ability to degrade NP was determined. The screening of microorganisms in river water and soil led to a collection of 23 strains of bacteria and five strains of fungi. Two strains of bacteria, identified as Pseudomonas sp. and Acidovorax sp., possessed great ability for degrading NP. The NP degradation rate of Pseudomonas sp. did not change with the NP concentration (50-100mg/L) . In contrast, the NP degradation rate of Acidovorax sp. increased with increasing NP concentration. Acidovorax sp. possessed the greatest NP degradation activity at 35°C. No NP degradation activity was observed for Pseudomonas sp. at temperatures higher than 30°C. Even when non-NP carbon sources such as glucose or sucrose were added, the NP degradation rates for both bacteria did not decrease. In addition, the estrogenic activity of NP decreased depending on the amount of NP residues determined by the yeast two-hybrid system.

Nadifloxacin, an antiacne quinolone antimicrobial, inhibits the production of proinflammatory cytokines by human peripheral blood mononuclear cells and normal human keratinocytes.

BACKGROUND: Acne vulgaris is a chronic inflammatory disease involving colonization of Propionibacterium acnes (P. acnes), activation of neutrophils and lymphocytes. Circumstantial evidence suggests that antigen-independent and -dependent immune responses against P. acnes are involved in the pathogenesis of inflammatory acne. Epidermal keratinocytes are also suggested to be involved in initiation and progression of cutaneous inflammation. Nadifloxacin, a fluorinated quinolone, has potent antimicrobial activities against Gram-negative and -positive microbes and is used to treat multiple inflamed acne lesions. However, its effect on immune conferring cells such as mononuclear cells and keratinocytes has not been examined. OBJECTIVE: To evaluate the possible involvement of potential anti-inflammatory activity of nadifloxacin in its therapeutic effect on inflammatory acne, we examined the effects of nadifloxacin, in comparison with other antibiotics used to treat acne vulgaris, on cytokine production by human peripheral blood mononuclear cells (PBMC) and keratinocytes. METHODS: Cytokine production by PBMC was determined after treatment with heat-killed P. acnes in the presence or absence of antimicrobials using a real-time PCR and ELISA. Cultured human epidermal keratinocytes were stimulated by IFN-gamma plus IL-1beta and the effects of antimicrobials were examined by using ELISA. RESULTS: Nadifloxacin as well as macrolide antibiotics and clindamycin inhibited IL-12 and IFN-gamma production by PBMC stimulated by heat-killed P. acnes. The drug also inhibited the IL-1alpha, Il-6, IL-8 and GM-CMS production by keratinocytes treated with IFN-gamma plus IL-1beta. CONCLUSIONS: Inhibitory effects of nadifloxacin to activate T cells and keratinocytes may be involved at least in part in the mechanism of its therapeutic effect against inflammatory acne.

Synergistic antiviral effect of a combination of mouse interferon-alpha and interferon-gamma on mouse hepatitis virus.

Although interferon (IFN)-alpha and IFN-gamma have been reported to exhibit a synergistic antiviral effect through the different signaling pathways in vitro, their therapeutic efficacy is not well defined in vivo. The current study was carried out to investigate the combined antiviral effect in a model of mouse hepatitis virus Type 2 (MHV-2) infection, in which fulminant hepatitis is developed. MHV-2 was injected intraperitoneally into 4-week-old ICR mice, IFN or the vehicle was administered intramuscularly for 5 days, and the antiviral effect was evaluated based on survival periods, liver histology, serum alanine transaminase (ALT) levels, and MHV-2 virus titers in the liver tissues. The animals in the group treated with a combination of IFN-alpha and IFN-gamma survived for longer periods than the groups treated with IFN-alpha alone and IFN-gamma alone (IFN-alpha 10(3) (IU/mouse)/-gamma 10(3) vs. IFN-alpha 10(3), P < 0.005; IFN-alpha 10(3)/-gamma 10(3) vs. IFN-gamma 10(3), P < 0.001). This is consistent with the lower levels of hepatocellular necrosis and serum ALT and the decreased titers of MHV-2 virus in the liver tissues (48 hr, P < 0.001; 72 hr, P < 0.001). These findings indicate that a combination of IFN-alpha and IFN-gamma exhibits a synergistic antiviral effect on MHV-2 infection. The biology of MHV-2 is quite different from that of human hepatitis viruses; however, these results suggest the beneficial combined therapy of IFN-alpha and IFN-gamma for the treatment of human viral hepatitis.