Povidone-iodine (PVP-I) is used for infection control and preoperative sterilization of the oral and pharyngeal regions. Marketed preparations containing cetylpyridinium chloride (CPC) are used to inhibit growth of oral bacteria. We conducted an in vitro study of the sterilizing effects of these microbicides on 10 oral bacterial strains and fungi related to pneumonia and periodontal disease, after dilution with phosphate-buffered saline (PBS), saliva, and components in saliva. The CPC solution was evaluated at 50 mg/100 mL, which is the concentration used in products. CPC sterilized all strains within 1 minute. Prolongation of the sterilization time associated with dilution was more gradual in comparison to PVP-I solution. CPC sterilized 7 of 10 microbial strains within 3 minutes at 3 mg/100 mL. At 500 mg/100 mL, which is near the upper limit of the concentration that is actually used, PVP-I solution sterilized 7 microbial strains within 3 minutes. However, PVP-I had no sterilization effect when diluted to 100 mg/100 mL or lower. With addition of saliva, PVP-I sterilized 2 microbial strains within 3 minutes at 500 mg/100 mL, whereas CPC solution sterilized 9 microbial strains within 1 minute at 50 mg/100 mL. Our results show that in use influenced by dilution with saliva, CPC is likely to maintain a strong sterilization effect, whereas PVP-I may have a reduced effect.
Escherichia coli cells were suspended in phosphate-buffered saline solutions (pH 7.4) at physiological (0.9 %) and hyperosmotic (3.5, 5.0, and 10.0 %) concentrations of sodium chloride (NaCl) and stored at 5, 10, 15, 20, and 25 °C up to 48 d. During storage at 5 and 10 °C, viable cell counts decreased approximately from 9 log CFU/ml to 6-7 log CFU/ml, and NaCl showed slight protective effect on the decrease. When stored at 15, 20, and 25 °C, the counts decreased with increases in NaCl concentration and/or storage temperature. The cells in 10.0 % NaCl suspension became nondetectable after storage at 25 °C for 28 d. Under some storage conditions (NaCl ≤ 5 %, 20 and 25 °C), the counts approached constant values, indicating possible adaptation to NaCl. Injured cells were observed at 5.0 and 10.0 % NaCl. However, recovery was observed only at 5.0 % NaCl during storage at 20 °C. In addition, more cells were detected on nonselective medium when incubated at 37 °C than at 25 °C. Higher hyperosmotic NaCl solutions at higher storage temperatures reduced more viable cells of E. coli.
House dust mites, Dermatophagoides pteronyssinus are present in the indoor environments, such as pillows and carpets. In this study, we investigated the mite control effect of branched chain fatty acids (2-ethylhexanoic acid (iso-C8), 2-butyloctanoic acid (iso-C12), isopalmitic acid (iso-C16) and their mechanism of action. These fatty acids showed a higher acaricidal activity than the straight chain fatty acids. Among these, iso-C12 had the highest acaricidal activity (LC50: 13mM) and more than 50% repellence rate at 0.20% (8.0mM) concentration. In the fumigant mortality bioassay, iso-C8 was 4 times more toxic than iso-C12 and isoC-16 in the gas phase. However, all branched chain fatty acids showed higher acaricidal activities on direct contact than fumigation. As the concentration of these fatty acids decreased, the number of deaths decreased and the number of escapes increased. There was no significant change in the mite epidermis due to contact with any of the fatty acids used. All branched chain fatty acids immobilized more than half of the mites within 90min of exposure. These results were consistent with the tendency of immobilizer type miticides targeting the respiratory system.
Photosynthetic bacterium (PSB) was isolated from sediment samples of Yamagawa Bay, Kagoshima, Japan. Phylogenetic analysis results of PSB isolate were closely related to Rhodobacter sphaeroides, purple non-sulfur photosynthetic bacteria (PNSB). Pink-colored smooth edges of single bacterial colonies were observed after 3-5 days of incubation period on Basic I medium agar plates. Rhodobacter sphaeroides microscopic examination showed a short rod cell (1-2 µm length) with round ends. Sediment and water samples used for ciliates cultivation were collected from Kuwano-ura Bay, Koshiki Island, Japan. Ciliates were cultivated using fish meal with radish leaves medium (MI), with sediment into MI (MII) and algae media (MIII). The use of the algae media (MIII) in cultivation mixture produced the highest total number of ciliates. Big size ciliates were identified as Euplotes minuta and Cyclidium varibonneti, while small size was identified as Micrometopion nutans, based on PCR-DGGE. When ciliates were cultured with the PSB isolate, Rhodobacter sphaeroides as a feed, ciliates grow to 2,081 individual ml-1 72 hrs later. These ﬁndings indicate that PNSB can be used to promote ciliates growth.
Rapid microbiological methods (RMMs) have been used as novel quality control technologies in industry. The ability of RMMs to detect stressed bacteria, in particular, is of continued interest due to the limitations of the conventional method in stressed bacteria detection. Accordingly, there is a need to better characterize an RMM’s ability to detect stressed microorganisms. Previously we reported on the detection ability of an intrinsic fluorescence-based RMM using a 50% injured (determined based on colony-forming ability) bacterial cell group after heat treatment at 55°C for 8 min. In this study, we added further information about the physiological state of the heat treated Escherichia coli, besides proliferation ability, by investigating respiratory activity using CTC fluorescent staining and expression of DnaK, a heat shock protein. It was found that 89% of cells (control 96%) retained respiratory activity, but only 20% (control 41%) retained proliferation ability after heat treatment. The difference between the percentage of cells with respiratory activity versus that of cells still capable of proliferation further supports the existence of viable but non-culturable stressed cells in the test sample. Also, we suggest such analysis would be one approach to confirming the use of stressed as opposed to dead cells when evaluating an RMM’s ability to detect stressed microorganisms.
Acanthamoeba is one of the organisms that cause corneal infection. In this study, attention was focused on potassium isostearate (iso-C18K, a branched chain fatty acid salt) for use in a multipurpose solution (MPS) against Acanthamoeba. An anti-amoebic test against Acanthamoeba castellanii ATCC 30010 (trophozoites type) was conducted. As a result, a growth reduction effect of 4 log units (99.99% suppression) was observed after incubation with 150 mM (5.0 w/v%) iso-C18K for 10 minutes. Furthermore, after the amoeba suspension was mixed with iso-C18K, disruption of cell membranes were observed, and the minimum amoebacidal concentration (MAC) at that time was 9.6 mM (0.31 w/v%). To evaluate the effectiveness as an MPS, assessment by verification tests was conducted using contact lenses. Reducing the concentration of iso-C18K caused a decrease in the number of viable cells, which was confirmed at a MAC of 1.2 mM (0.039 w/v%).
In this study, lactic acid bacteria (LAB) strains were isolated from ground beef, and it was analyzed if they have any effect on the growth of two reference bacteria (Salmonella sp. and Escherichia coli). It was found that five isolates showed an inhibitory effect in both reference bacteria by spot at the lawn assay. These bacteria were selected to perform growth kinetics in co-culture to determine if they modify the growth parameters of the reference bacteria. Subsequently, LAB cultures and three treatments (crude extract, thermally treated and thermally treated with neutral pH) of cells free supernatants (CFS) were screened by the agar well diffusion assay. In co-culture, selected LAB altered the growth rate and reduce the maximum population of both reference bacteria. While, LAB cultures and CFS also showed antimicrobial activity, and there was no significant difference among CFS treatments. LAB isolated from ground beef showed an antimicrobial effect against the reference bacteria that could be used for meat biopreservation purposes.
Aspergillus section Versicolores species, except Aspergillus sydowii, produce a carcinogenic mycotoxin sterigmatocystin (STC). Since these fungi are found in varied environmental milieu including indoor dust and food products, our aim was to develop a sensitive and convenient assay to detect STC producing fungal strains. We made use of a high discrimination DNA polymerase (HiDi DNA polymerase), for single nucleotide polymorphism (SNP)-based PCR amplification. Using specific primer pairs based on the SNPs between A. sydowii and other strains of Aspergillus section Versicolores, we succeeded in amplifying the genomic DNA all target strains except A. sydowii. These results confirm that the SNP-based PCR amplification technique, using a high discrimination DNA polymerase, was a reliable and robust screening method for target fungal strains.