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Antibiotic-Resistant Bacterial Group in Field Soil Evaluated by a Newly Developed Method Based on Restriction Fragment Length Polymorphism Analysis

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Advances in Microbiology 2015年第12期5卷 807-816页

作者： Katsuji Watanabe naoto horinishi Kunimasa Matumoto Department of Life Environment and Materials Science Fukuoka Institute of Technology Fukuoka Japan

Spreading of antibiotic resistant bacteria into environment is becoming a major public health problem, implicating affair of the indirect transmission of antibiotic resistant bacteria to human through drinking water, or vegetables, or daily products. Until now, the risk of nosocomial infection of antibiotic resistant bacteria has mainly been evaluated using clinical isolates by phenotypic method. To evaluate a risk of community-acquired infection of antibiotic resistant bacteria, a new method has been developed based on PCR-RFLP without isolation. By comparing restriction fragment lengths of the 16S rDNA gene from bacterial mixture grown under antibiotic treatment to those simulated from the DNA sequence, bacterial taxonomies were elucidated using the method of Okuda and Watanabe [1] [2]. In this study, taxonomies of polymyxin B resistant bacteria group in field soils, paddy field with organic manure and upland field without organic manure were estimated without isolation. In the both field soils, the major bacteria grown under the antibiotic were B. cereus group, which had natural resistance to this antibiotic. In field applied with organic manure, Prevotella spp., and the other Cytophagales, which were suggested to be of feces origin and to acquire resistance to the antibiotic, were detected. When numbers of each bacterial group were roughly estimated by the most probable number method, B. cereus group was enumerated to be 3.30 × 106 MPN/g dry soil in paddy field soil and 1.32 × 106 MPN/g dry soil in upland filed. Prevotella spp. and the other Cytophagales in paddy field were enumerated to be 1.31 × 106 MPN, and 1.07 × 106 MPN·g-1 dry soil.

关键词： Polymyxin B Resistant Bacteria Field Soil Microchip Electrophoresis Multiple Enzyme Restriction Fragment Length Polymorphism Analysis the Most Probable Number Method

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A Simple Evaluation System for Microbial Property in Soil and Manure

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Advances in Microbiology 2016年第2期6卷 88-97页

作者： naoto horinishi Kunimasa Matsumoto Katsuji Watanabe Department of Life Environment and Materials Science Graduate School of Engineering Fukuoka Institute of Technology Fukuoka Japan

Analyses of microbial properties in soil and manure had always included the problem that there was no available standard method to evaluate microbial property. The one of the major problems was the vast diversity and the enormous population of soil microorganisms [1], the other was an existence of numerically dominant unculturable microorganisms which comprise 99% of soil habitat [2]. We evaluated whether our newly developed method, by which taxonomies and their number of each bacterial groups were estimated, could be used as evaluation method of microbial properties of soils and manures. In the forest soil, β-Proteobacteria, which included Burkholderia sp., Ralstonia sp., and Alcaligenes sp., was numerically dominant bacteria (3.64 × 106 MPN g-1 dry soil), followed by γ-Proteobacteria (1.32 × 106 MPN), δ-Proteobacteria (0.006 × 106 MPN), and the other gram negative bacteria (0.006 × 106 MPN). In the commercial manure, Actinobacteria, which included Streptoverticillium salmonis, Mycrococcus sp., Streptomyces bikiniensis, and Microbacterium ulmi, was numerically dominant bacterial group (30.8 × 106 MPN), followed by α-Proteobacteria (26.0 × 106 MPN), β-Proteobacteria (17.1 × 106 MPN), δ-Proteobacteria (11.2 × 106 MPN), the other Firmicutes (1.71 × 106 MPN), γ-Proteobacteria (0.5 × 106 MPN), and the other gram negative bacteria (0.05 × 106 MPN). In the upland field, the other Firmicutes, which included Paenibacillus sp., was numerically dominant bacteria (4.41 × 106 MPN), followed by Actinobacteria (2.14 × 106 MPN), Bacillus sp. (2.14 × 106 MPN), and γ-Proteobacteria (0.35 × 106 MPN). Although the precision of the affiliations became lower because of higher diversity of samples and the number of some Antinobacteria and Firmicutes might be underestimated by the used PCR condition, the method was

关键词： Evaluation System Microbial Property Soil and Manure Multiple Enzyme Restriction Fragment Length Polymorphism Analysis The Most Probable Number Method Microchip Electrophoresis

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Identification and Enumeration Method of Both Eukaryotic and Prokaryotic Microorganisms in Food Sample

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Food and Nutrition Sciences 2016年第5期7卷 345-354页

作者： Katsuji Watanabe naoto horinishi Kunimasa Matsumoto Yuji Sogabe Department of Life Environment and Materials Science Graduate School of Engineering Fukuoka Institute of Technology Fukuoka Japan Global Application Development Center Shimadzu Co. Kyoto Japan

The method to analyze both eukaryotic and prokaryotic microorganisms without preliminary microbial information of sample seemed to be useful not only for research and investigation of microorganisms but also for industry using microorganisms. In the present manuscript, preparation of a new DNA primers, new reference database for 18S rDNA for our newly developed method [1]- [3], and analyses of eukaryotic and prokaryotic microorganisms in fermentation products were presented. In komekouji, Aspergillus spp., was enumerated to be 46.5 × 106 MPN g-1, and Penicillium spp., was enumerated to be 1.5 × 106 MPN g-1. In dry yeast, Saccharomyces group, were enumerated to be 8600 × 106 MPN g-1. In komekouji-miso, no eukaryotic microorganism was detected, while the other Bacillus spp., was numerically dominant (21.5 × 106 MPN g-1) as prokaryotic microorganisms, followed by B. subtilis group (4.65 × 106 MPN g-1), and the other Firmicutes (3.7 × 106 MPN g-1). The komekouji-miso included lower number of Actinobacteria (0.15 × 106 MPN g-1), Burkhokderia sp. (1.5 × 106 MPN g-1), and the other α,β,γ-proteobacteria (0.12 × 106 MPN g-1). In sake-kasu, both prokaryote and eukaryote were not detected by the method. Present results indicated that using both universal primers for eukaryotic and prokaryotic microorganisms, each groups of prokaryotic and eukaryotic microorganisms were enumerated without any preliminary information nor setting up standard curve, which were required for real time PCR.

关键词： Eukaryotic Microorganisms Prokaryotic Microorganisms Multiple Enzyme Restriction Fragment Length Polymorphism Analysis the Most Probable Number Method Microchip Electrophoresis

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Evaluation of the Method Based on Restriction Fragment Length Polymorphism Analysis as Simple Analysis Method of Lactic Acid Bacteria in Foods

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Food and Nutrition Sciences 2016年第3期7卷 163-172页

作者： Kunimasa Matsumoto Kouya Shimada naoto horinishi Katsuji Watanabe Department of Life Environment and Materials Science Graduate School of Engineering Fukuoka Institute of Technology Fukuoka Japan Department of Life Environment and Materials Science Faculty of Engineering Fukuoka Institute of Technology Fukuoka Japan

Lactic acid bacteria have not only been used to produce various kinds of fermented food, but also used as probiotic products. As lactic acid bacterial group was consisted from diverse genera, a simple inspection method by which numbers and contained microorganisms could be automatically analyzed without any preliminary information was required to use them more effectively. In this manuscript, lactic acid bacterial groups in commercial products of kimuchi, komekouji-miso, and yoghurt were identified and enumerated by our newly developed method [1]-[3], to evaluate whether the method could be used as an inspection method of various food samples. In kimuchi, numerically dominant bacteria were Lactobacillus sakei, and L. casei (1.4 × 104 MPN g-1) and Leuconostoc spp. (l.4 × 104 MPN). In kouji-miso, numerically dominant bacteria was Bacillus spp. (3 × 103 MPN), which mainly included B. subtilis group and B. cereus group. Lactic acid bacteria such as Lactobacillus spp., or Lactococcus spp., included in the komekouji-miso, could be enumerated after 3 days incubation (1.24 × 104 MPN), but not detected after 7 days incubation. In yoghurt A and C, Lactococcus lactis was detected as numerically dominant lactic acid bacteria (3.0 × 105 MPN). In yoghurt B, Lactobacillus spp., or Lactococcus spp., was detected not only by a culturebased method but also by an unculture-based method, although there was a difference between the both estimated numbers. The present results suggested that the method might become useful as a simple inspection method of food microorganisms, because time and labor of the analysis could be reduced by using an unculture-based method and MCE-202 MultiNA. In this study, Bifidobacteriium spp. was not detected in B and C yoghurt, in spite of indicating their existence, and numbers of lactic acid bacteria were lower than the level of the daily product regulation, because 16S rDNA of Bifidobacteriium spp. might not be amplified by the used PCR conditio

关键词： Multiple Enzyme Restriction Fragment Length Polymorphism Analysis Most Probable Number Method Lactic Acid Bacteria Komekouji-Miso Kimuchi Yoghurt

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A New Evaluation Method for Antibiotic-Resistant Bacterial Groups in Environment

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Advances in Microbiology 2016年第3期6卷 133-151页

作者： Katsuji Watanabe naoto horinishi Kunimasa Matsumoto Akihiro Tanaka Kenichi Yakushido Department of Life Environment and Materials Science Fukuoka Institute of technology Fukuoka Japan National Agricultural Research Center for Kyushu-Okinawa Region National Agriculture and Food Research Organization Kumamoto Japan National Agricultural Research Center National Agriculture and Food Research Organization Tsukuba Japan

In the present manuscript it was presented whether spreading of antibiotic resistant bacterial groups in environment could be monitored by our newly developed method by enumerating antibiotic resistant bacterial groups in various biological wastes and composts. Although the numbers were not so high, diverse kinds of colistin resistant bacteria (25 mg·L-1) were included in row cattle feces (1.78 × 104 MPN g-1) and cattle feces manure (>3.84 × 104 MPN g-1). Compost originated from leftover food (>44.8 × 104 MPN g-1) and shochu lee (>320 × 104 MPN g-1) included higher numbers of chlortetracycline resistant Pseudomonas sp., (25 mg·L-1), and row cattle feces included higher numbers of chlortetracycline resistant Enterobacteriacea (15.7 × 104 MPN g-1), which mostly consisted from Pantoea sp. or Xenorhobdus doucetiae. Numbers of multi drug resistant bacteria, resistant to 25 mg·L-1 of ciprofloxacin, streptomycin, chloramphenicol, and ampicillin, were the highest in row cattle feces (>143.6 × 104 MPN g-1), followed by cattle feces manure (4.19 × 104 MPN g-1), and shochu lee (0.36 × 104 MPN g-1), which included diverse kinds of bacterial group. The present results indicated that higher numbers of multi drug resistant bacteria were typically found in row cattle feces, and the method was found suitable to enumerate and identify them. These results suggested that the method might become their environmental risk evaluation method.

关键词： Colistin Resistant Bacteria Chlortetracycline Resistant Bacteria Multi Drug Resistant Bacteria Multiple Enzyme Restriction Fragment Length Polymorphism Analysis The Most Probable Number Method Microchip Electrophoresis System

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Bacterial Groups Concerned with Maturing Process in Manure Production Analyzed by a Method Based on Restriction Fragment Length Polymorphism Analysis

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Advances in Microbiology 2015年第13期5卷 832-841页

作者： Katsuji Watanabe naoto horinishi Kunimasa Matumoto Akihiro Tanaka Kenichi Yakushido Department of Life Environment and Materials Science Fukuoka Institute of Technology Fukuoka Japan National Agricultural Research Center for Kyushu-Okinawa Region National Agriculture and Food Research Organization Koshi Japan National Agricultural Research Center National Agriculture and Food Research Organization Tsukuba Japan

Composting is a biological aerobic decomposition process consisted from different phases. Although the Japanese Standards for manure recommended that it took at least 6 months to complete the maturing phase, there was no reliable ground. In order to find out shortening method of the maturing phase, the microorganisms concerned with a progress of the maturing was determined by using the most probable number method (MPN) and PCR-RFLP of the 16S rDNA, which was found effective to provide numbers and taxonomy of polymyxin B resistant bacterial groups in the former paper [1]. Compared to the numbers after thermophilic phase, those of Actinobacteria, δ-proteobacteria, and the other gram negative bacteria increased to 50 times, 20 times, and 105 times, respectively, after maturing phase, while those of Bacillus spp., and α and β-proteobacteria decreased to 1/10, and 1/105 after maturing phase. Numbers of the other Fumicutes, and γ-proteobacteria remained in the same revel. Actinobacteria, δ-proteobacteria, and the other gram negative bacteria might be concerned with a progress of the maturing phase, because these bacterial groups were detected and enumerated due to their proliferation ability. Although number of Acitinobacteria might be underestimated because of a PCR bias, the method was found effective for the purpose to monitor bacteria actively proliferated in culture medium.

关键词： Maturing Phase Manure Production Microchip Electrophoresis Multiple Enzyme Restriction Fragment Length Polymorphism Analysis The Most Probable Number Method

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Loess magnetic properties in the Ili Basin and their correlation with the Chinese Loess Plateau

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Science China Earth Sciences 2010年第3期53卷 419-431页

作者： naoto Ishikawa Graduate School of Human and Environmental Studies Kyoto University

Over the past two decades, magnetoclimatological studies of loess-paleosol sequences in the Chinese Loess Plateau (CLP) have made outstanding achievements, which greatly promote the understanding of East Asian paleomonsoon evolution, inland aridification of Asia, and past global climate changes. Loess magnetic properties of the CLP have been well studied. In contrast, loess magnetic properties from outside the CLP in China have not been fully understood. We have little knowledge about the magnetic properties of loess in the Ili Basin, an intermontane depression of the Tianshan (or Tien Shan) Mountains. Here, we present the results of rock magnetic measurements of the Ili loess including mass magnetic susceptibility (χ) and anhysteretic remanent magnetization (ARM), high/low temperature dependence of susceptibility (TDS) and hysteresis, as well as X-ray diffraction (XRD) for mineral analysis. Based on the comparison with loess-paleosol sequences in the CLP (hereafter referred to as the Chinese loess), we discuss the possible magnetic susceptibility enhancement mechanism of the Ili loess. The results show that 1) the total magnetic mineral concentration of the Ili loess is far lower than that of the Chinese loess, though they have similar magnetic mineral compositions. The ferrimagnetic minerals in the Ili loess are magnetite and maghemite, and the antiferromagnetic mineral is hematite; XRD analysis also identifies the presence of ilmenite. The ratio of maghemite is lower in the Ili loess than in the Chinese loess, but the ratios of magnetite and hematite are higher in the Ili loess than in the Chinese loess. 2) The granularity of magnetic minerals in the Ili loess, dominated by pseudo-single domain (PSD) and multi-domain (MD) grains, is generally much coarser than that of the Chinese loess. Ultrafine pedogenically-produced magnetic grains have a very limited contribution to the susceptibility enhancement. Rather, PSD and MD particles of magnetite and maghemite are th

关键词： loess magnetic property magnetic susceptibility the Ili Basin the Chinese Loess Plateau

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Estimating camera parameters from starry night photographs

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Computational Visual Media 2020年第4期6卷 445-454页

作者： naoto Ishikawa Yoshinori Dobashi Graduate School of Information Science and Technology Hokkaido UniversitySapporoJapan

We propose an efficient, specific method for estimating camera parameters from a single starry night image. Such an image consists of a collection of disks representing stars, so traditional estimation methods for common pictures do not work. Our method uses a database, a star catalog, that stores the positions of stars on the celestial sphere. Our method computes magnitudes(i.e., brightnesses) of stars in the input image and uses them to find the corresponding stars in the star catalog. Camera parameters can then be estimated by a simple geometric calculation. Our method is over ten times faster and more accurate than a previous method.

关键词： astrophotography constellation star identification pattern matching

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High-Resolution Spectroscopy of Winds Associated with T Tauri Stars

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Research in Astronomy and Astrophysics 2016年第2期16卷 89-96页

作者： naoto Iguchi Yoichi Itoh Graduate School of Science Kobe University1-1 Rokkodai-choNada-kuKobeHyogo 657-8501Japan Nishi-Harima Astronomical Observatory Center for AstronomyUniversity of Hyogo407-2NishigaichiSayoHyogo 679-5313Japan

We carried out optical high-resolution spectroscopy of T Tauri stars using the Subaru Telescope. Using archived data from the Keck Telescope and the Very Large Telescope, we detected forbidden lines of [S II] at 4069A, in addition to those of [O I] at 5577 A and 6300A, for 13 T Tauri stars. We consider that low-velocity components of these forbidden lines emanate from the wind associated with T Tauri stars. Using two flux ratios of the three lines, we simultaneously determined the hydrogen density and temperature of the winds. The winds ofT Tauri stars have a hydrogen density of 2.5 × 106 cm^-3 - 2.5 × 10^9 cm^-3 and a temperature of 10 800 -18 000 K. The mass loss rates by the wind are estimated to lie in the range from 2.0 × 10^-10 M⊙ yr-1 to 1.4× 10^-9 M⊙ yr-1. The mass loss rates are found to increase with increasing mass accretion rates. The ratio of the mass loss rate to the mass accretion rate is 0.001-0.1 for classical T Tauri stars and 0.1-1 for transitional disk objects.

关键词： stars pre-main sequence -- stars emission lines

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Numerical Simulation of Tip Leakage Vortex Effect on Hydrogen-Combustion Flow around 3D Turbine Blade

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Journal of Thermal Science 2008年第2期17卷 186-192页

作者： naoto Miyama Kazuaki Inaba Makoto Yamamoto Department of Mechanical Engineering Tokyo University of Science Graduate Aeronautical Laboratories California Institute of Technology

In these years, a lot of environmental problems such as air pollution and exhaustion of fossil fuels have been discussed intensively. In our laboratory, a hydrogen-fueled propulsion system has been researched as an alternative to conventional systems. A hydrogen-fueled propulsion system is expected to have higher power, lighter weight and lower emissions. However, for the practical use, there exist many problems that must be overcome. Considering these backgrounds, jet engines with hydrogen-fueled combustion within a turbine blade passage have been studied. Although some studies have been made on injecting and burning hydrogen fuel from a stator surface, little is known about the interaction between a tip leakage vortex near the suction side of a rotor tip and hydrogen-fueled combustion. The purpose of this study is to clarify the influence of the tip leakage vortex on the characteristics of the 3-dimensional flow field with hydrogen-fueled combustion within a turbine blade passage. Reynolds-averaged compressible Navier-Stokes equations are solved with incorporating a k-ε turbulence and a reduced chemical mechanism models. Using the computational results, the 3-dimensional turbulent flow field with chemical reactions is numerically visualized, and the three-dimensional turbulent flow fields with hydrogen combustion and the structure of the tip leakage vortex are investigated.

关键词： numerical simulation CFD gas turbine hydrogen combustion tip leakage vortex

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