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Interpreting the Results of the Conventional Plate Culture and Gene Detection Methods for Legionella Detection in Environmental Water Samples. 环境水样中军团菌常规平板培养和基因检测方法结果的解释。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.121
Hiroaki Inoue

The conventional plate culture method is widely used as a method for detection of Legionella in environmental water samples, but to obtain results takes more than a week. Because it is much quicker, the gene detection method has become widespread as an alternative detection method. However, the results of gene detection and plate culture methods may differ even when the same sample is examined; the gene detection method shows a higher detection ratio than the plate culture method. The reason for this difference is that the plate culture method detects Legionella cells that have the ability to form colonies on an agar plate, whereas the gene detection method detects any Legionella genes present regardless of the state of the Legionella. In this paper, we consider the factors that cause differences between the results of the plate culture and gene detection methods, and how to interpret the results of each.

传统的平板培养法被广泛用于环境水样中军团菌的检测,但获得结果需要一周以上的时间。由于其速度快得多,基因检测法作为一种替代检测方法已得到广泛应用。然而,基因检测和平板培养方法的结果可能不同,即使在检查相同的样品;基因检测法比平板培养法的检出率更高。造成这种差异的原因是,平板培养方法检测的是有能力在琼脂平板上形成菌落的军团菌细胞,而基因检测方法检测的是任何军团菌基因,而不管军团菌的状态如何。在本文中,我们考虑了导致平板培养和基因检测方法结果差异的因素,以及如何解释每种结果。
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引用次数: 0
The Effects of Peptidoglycan on the Photocatalytic Bactericidal Activity of Titanium Dioxide. 肽聚糖对二氧化钛光催化杀菌活性的影响。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.167
Ayano Takao, Tomonori Suzuki

For the elucidation of the mechanism underlying the photocatalytic bactericidal activity of titanium dioxide (TiO2), we focused on the peptidoglycan layer, a component of the bacterial cell wall. The effect of this layer on the photocatalytic bactericidal activity of TiO2 was evaluated by determining the survival rates of Lactobacillus plantarum (intact cells) and its protoplast cells. Mesoplasma florum, which does not originally possess the peptidoglycan layer, was also used. Our results revealed that the survival rates of the intact cells were lower than those of the protoplast cells. In addition, there was no significance between the survival rates of M. florum cells and the protoplast cells of L. plantarum. It was suggested that the presence of the peptidoglycan layer increases the bactericidal effect by the photocatalysis.

为了阐明二氧化钛(TiO2)光催化杀菌活性的机制,我们重点研究了细菌细胞壁的一种成分肽聚糖层。通过测定植物乳杆菌(完整细胞)及其原生质体细胞的存活率来评估该层对TiO2光催化杀菌活性的影响。本研究还使用了原本不具有肽聚糖层的花中原体。结果表明,完整细胞的存活率低于原生质体细胞的存活率。此外,花芽孢杆菌细胞的存活率与植物芽孢杆菌原生质体细胞的存活率无显著性差异。结果表明,肽聚糖层的存在增加了光催化杀菌效果。
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引用次数: 2
Analysis of Fungal Flora in the Dust of Bedding in Japanese Houses and Genetic Identification of Yeasts Isolated from the Dust. 日本住宅床上用品粉尘中真菌区系分析及分离酵母的遗传鉴定。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.193
Kazuhiro Hashimoto, Fumi Yamazaki, Noriko Kohyama, Yuji Kawakami

This study examined the fungal flora contained in the dust of bedding used in 50 houses in Japan. The result showed that the mycoflora having the largest isolation rate was yeasts, which were isolated by 42 out of 50 houses (84%), and exceeded the isolation rate of Cladosporium spp. (80%) and Aspergillus spp. (66%). In addition, the isolation rate of Alternaria, which was an important fungus causing asthma, 66% was being considered as a high isolation rate, and this result was very interesting. The isolation rate of xerophilic fungi such as Aspergillus restrictus and Wallemia often found in house dust on the floor, was not very high. Forty-one strains of yeasts isolated from each dust sample were identified, and Naganishia diffluens species complex and Filobasidium magnum had a larger number of 13 strains, respectively. Since N. diffluens was the yeasts often isolated from human skin, it was thought to be an association between the fungal skin flora and fungal flora of bed dust. Meanwhile, there was no report of isolation of F. magnum from house dust previously. To the best of our knowledge, this is the first study showing its isolation from bedding with relatively high frequency.

本研究检测了日本50个家庭使用的床上用品粉尘中含有的真菌菌群。结果表明,真菌菌群分离率最高的是酵母菌,50个菌群中有42个(84%)分离出酵母菌,超过了枝孢杆菌(80%)和曲霉菌(66%)的分离率。此外,哮喘重要真菌Alternaria的分离率高达66%,被认为是很高的分离率,这一结果非常有趣。室内地板灰尘中常见的限制性曲霉、瓦勒菌等嗜干真菌的分离率不高。从每个粉尘样品中分离到41株酵母菌,其中naganishi diffluens种复合体和Filobasidium magnum菌数最多,分别为13株。由于散流奈索菌是通常从人类皮肤中分离出来的酵母菌,因此它被认为是皮肤真菌菌群与床尘真菌菌群之间的联系。同时,尚无从室内粉尘中分离到该菌的报道。据我们所知,这是第一次研究表明它与相对高频率的床上物隔离。
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引用次数: 6
Effect of Hyperosmotic Salt Concentration and Temperature on Viability of Escherichia coli during Cold Storage. 高渗盐浓度和温度对冷藏过程中大肠杆菌活力的影响。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.55
Xue Zhang, Yoshiko Nakaura, Junzhang Zhu, Zhenya Zhang, Kazutaka Yamamoto

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.

将大肠杆菌细胞悬浮在生理(0.9%)和高渗(3.5、5.0和10.0%)氯化钠(NaCl)浓度的磷酸盐缓冲盐水溶液(pH 7.4)中,并在5、10、15、20和25°C下储存48 d。在5和10°C下储存期间,活细胞计数约从9 log CFU/ml减少到6-7 log CFU/ml, NaCl对减少有轻微的保护作用。在15℃、20℃和25℃的贮藏条件下,随着NaCl浓度和(或)贮藏温度的增加,细菌数量减少。10.0% NaCl悬浮液在25°C下保存28 d后,细胞已无法检测到。在某些储存条件下(NaCl≤5%、20和25°C),细胞计数接近恒定值,可能是对NaCl的适应。在5.0和10.0% NaCl下观察损伤细胞。但在20°C条件下,仅在5.0% NaCl条件下恢复。此外,在37°C非选择性培养基上比在25°C孵育时检测到更多的细胞。较高的高渗NaCl溶液在较高的储存温度下降低了大肠杆菌的活菌量。
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引用次数: 2
Mite control effect of branched chain fatty acid on the house dust mite Dermatophagoides pteronyssinus. 支链脂肪酸对房尘螨的防螨效果。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.63
Aki Maruoka, Toshinari Koda, Hiroshi Morita

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.

室内环境,如枕头和地毯中存在室内尘螨。本研究研究了支链脂肪酸(2-乙基己酸(isoc8)、2-丁酸(isoc12)、异铝酸(isoc16)的防螨效果及其作用机理。这些脂肪酸比直链脂肪酸具有更高的杀螨活性。其中,iso-C12的杀螨活性最高(LC50: 13mM),在0.20% (8.0mM)浓度下的驱螨率超过50%。在熏蒸剂死亡率生物测定中,iso-C8的气相毒性是iso-C12和iso- 16的4倍。而支链脂肪酸直接接触的杀螨活性均高于熏蒸处理。随着这些脂肪酸浓度的降低,死亡人数减少,逃逸人数增加。在螨虫表皮没有明显的变化,由于接触任何脂肪酸使用。所有支链脂肪酸在暴露后90min内固定了一半以上的螨虫。这些结果与固定化型杀螨剂以呼吸系统为靶点的趋势一致。
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引用次数: 0
Investigation into the Physiological State of Heat Stressed Escherichia coli Used in the Evaluation Testing of an Intrinsic Fluorescence-Based RMM. 热应激大肠杆菌生理状态的研究用于本征荧光基RMM评价试验。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.91
Kanami Irie, Allison Scott, Norio Hasegawa

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.

快速微生物学方法(RMMs)已成为一种新型的质量控制技术。由于传统方法在压力细菌检测中的局限性,RMMs检测压力细菌的能力尤其受到人们的持续关注。因此,有必要更好地表征RMM检测应激微生物的能力。在此之前,我们报道了在55°C加热8分钟后,使用50%损伤(根据集落形成能力确定)的细菌细胞组检测基于本特性荧光的RMM的能力。在本研究中,我们通过使用CTC荧光染色研究呼吸活性和DnaK(一种热休克蛋白)的表达,进一步了解了热处理后大肠杆菌的生理状态,以及增殖能力。结果发现,热处理后89%的细胞(对照组96%)保留了呼吸活性,而只有20%的细胞(对照组41%)保留了增殖能力。具有呼吸活性的细胞百分比与仍能增殖的细胞百分比之间的差异进一步支持了测试样本中存在可存活但不可培养的应激细胞。此外,我们建议在评估RMM检测应激微生物的能力时,这种分析将是确认使用应激细胞而不是死细胞的一种方法。
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引用次数: 1
Changes in the Survivability of Marine Vibrio sp. under Hyper KCl Stress in the Presence of Betaine as Well as with Exposure to 37ºC. 甜菜碱和37℃环境下高氯化钾胁迫下海洋弧菌存活能力的变化
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.17
Yue Yin, Haruo Mimura
Survivability at hyper KCl stress was examined at 30ºC and 37ºC in the presence and absence of an osmoprotectant by using resting cells prepared from marine Vibrio sp. grown at early stationary phase. Survivability was decided by counting colonies. The number of initial cells, 107.1 ± 0.2 (CFU·mL-1), was reduced to 105.1 ± 0.5 and < 101.0 (CFU·mL-1) at 30ºC and 37ºC, respectively, by the exposure of resting cells, that were prepared from cells grown for 8 h at 0.5 M NaCl at 30ºC, to 1.2 M KCl and 50 mM NaCl for 3 h. Betaine externally existed as a final concentration of 50 mM mitigated hyper KCl stress to the resting cells at 37ºC. The number of surviving cells was maintained 104.9 ± 0.3 (CFU·mL-1) when resting cells, 106.5 ± 0.1 (CFU·mL-1), that were prepared from pre-adapted cells to relatively high concentration of KCl in the growth for 10 h at 0.8 M KCl and 50 mM NaCl at 37ºC, were exposed to 1.2 M KCl, 50 mM NaCl, and 50 mM betaine at 37ºC for 3 h. The results indicate that osmoadaptation system(s) in resting cells is temperature sensitive and betaine functions to mitigate hyper KCl stress to the resting cells at 37ºC.
在30ºC和37ºC条件下,在存在和不存在渗透保护剂的情况下,研究了高氯化钾胁迫下的存活能力,方法是使用在早期固定阶段生长的海洋弧菌制备的静息细胞。生存能力是通过计算菌落来决定的。在30℃0.5 M NaCl条件下培养8 h的细胞,在30℃和37℃条件下培养至1.2 M KCl和50 mM NaCl条件下培养3 h后,初始细胞数(107.1±0.2 (CFU·mL-1))分别减少到105.1±0.5和< 101.0 (CFU·mL-1)。甜菜碱以50 mM的终浓度外源存在,减轻了37℃条件下静息细胞的高KCl胁迫。预适应细胞在较高KCl浓度下,在0.8 M KCl和50 mM NaCl条件下生长10 h后,在1.2 M KCl, 50 mM NaCl条件下,静息细胞数为106.5±0.1 (CFU·mL-1),存活细胞数维持在104.9±0.3 (CFU·mL-1)。结果表明,静息细胞的渗透适应系统具有温度敏感性,甜菜碱可减轻静息细胞在37℃下的高氯化钾胁迫。
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引用次数: 1
Evaluation of Legiolert for Quantification of Legionella pneumophila from Bath Water Samples. Legiolert用于洗澡水嗜肺军团菌定量的评价。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.179
Hiroaki Inoue, Masumi Baba, Seina Tayama

Testing for Legionella spp. in public bath water samples is regulated in Japan. In this study, we used a total of 132 public bath water samples to compare the performance of Legiolert® and the conventional plate culture method for the enumeration of Legionella pneumophila. When Legiolert and plate culturing were performed at the same detection limit, L. pneumophila was detected in 26.5% of 132 samples by Legiolert, while 12.9% contained Legionella spp. (11.4% contained L. pneumophila) based on the plate culture method. Moreover, results of 83.3% of the total samples were consistent between the two methods, meaning that they were both positive or both negative. In this study, we demonstrated that Legiolert is a simpler and more effective method of monitoring for L. pneumophila in bath water samples.

日本对公共洗澡水样本中的军团菌进行检测。本研究采用132份公共洗澡水样本,比较Legiolert®与传统平板培养法对嗜肺军团菌的计数效果。军团菌与平板培养在相同检出限下,132份样品中,军团菌检出率为26.5%,平板培养法检出军团菌12.9%,其中嗜肺乳杆菌11.4%。而且,83.3%的总样本结果在两种方法之间是一致的,即两者都是阳性或都是阴性。在这项研究中,我们证明了Legiolert是一种更简单、更有效的监测洗澡水样品中嗜肺乳杆菌的方法。
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引用次数: 8
Factors Affecting Reduction in Preservative Efficacy in Nonwoven Fabrics. 影响非织造布防腐效果降低的因素。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.149
Yuko Endo, Masumi Mune, Jun Usukura

Insufficient preservative efficacy leads to microbial contamination. Cosmetic-impregnated products composed of nonwoven fabrics, such as wipes and masks, can be contaminated with microbes owing to their special form. However, the reduction of preservative efficacy in cosmetic-impregnated products remains unverified. This study aimed to investigate whether preservative efficacy is reduced in nonwoven fabrics impregnated with a cosmetic liquid and the factors affecting this reduction. First, we evaluated the preservative efficacy of face wipes and confirmed that the preservative efficacy was reduced after impregnation of cosmetic liquids into nonwoven fabrics. We thus hypothesized that the adsorption of the antimicrobial components onto nonwoven fabrics decreases the preservative efficacy. Unexpectedly, the antimicrobial components were scarcely adsorbed onto the fabrics, while microbial growth activity was significantly increased on the fabrics, as determined through microbial calorimetry. Furthermore, the antibacterial effects were reduced in the nonwoven fabrics. These results indicate that the nonwoven fabrics enhanced microbial growth, thus decreasing the preservative efficacy. Our results provide novel insights into the microbial control of products composed of nonwoven fabrics.

保鲜效果不足导致微生物污染。湿巾、口罩等由非织造布构成的化妆品浸渍产品,由于其特殊的形态,有可能被微生物污染。然而,在化妆品浸渍产品中防腐剂功效的降低仍未得到证实。本研究旨在探讨化妆品液体是否会降低非织造布的防腐效果,以及影响这种降低的因素。首先,我们对湿巾的防腐效果进行了评价,确认化妆品液体浸渍在无纺布中后,湿巾的防腐效果降低。因此,我们假设抗菌成分在非织造布上的吸附会降低防腐效果。出乎意料的是,抗菌成分几乎没有被吸附到织物上,而微生物的生长活性却显著增加,这是通过微生物量热法测定的。此外,非织造布的抗菌效果也有所降低。这些结果表明,非织造布促进了微生物的生长,从而降低了保鲜效果。我们的结果提供了新的见解,微生物控制产品组成的非织造布。
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引用次数: 1
Diversity of the Photosynthetic Bacterial Communities in Highly Eutrophicated Yamagawa Bay Sediments. 高富营养化山川湾沉积物光合细菌群落的多样性。
IF 1.2 4区 农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2020-01-01 DOI: 10.4265/bio.25.25
Islam Teiba, Takeshi Yoshikawa, Suguru Okunishi, Makoto Ikenaga, Mohammed El Basuini, Hiroto Maeda

Yamagawa Bay, located in Ibusuki, Kagoshima Prefecture, Japan, is a geographically enclosed coastal marine inlet, and its deteriorating seabed sediments are under an anoxic, reductive, sulfide-rich condition. In order to gain insight into diversity of anoxygenic photosynthetic bacteria (AnPBs) and their ecophysiological roles in the sediments, three approaches were adopted: isolation of AnPBs, PCR-DGGE of 16S rDNA, and PCR-DGGE of pufM. Among the bacterial isolates, relatives of Rhodobacter sphaeroides were most dominant, possibly contributing to transforming organic pollutants in the sediments. Abundance of Chlorobium phaeobacteroides BS1 was suggested by 16S rDNA PCR-DGGE. It could reflect intensive stratification and resultant formation of the anoxic, sulfide-rich layer in addition to extreme low-light adaptation of this strain. Diverse purple non-sulfur or sulfur bacteria as well as aerobic anoxygenic photoheterotrophs were also detected by pufM PCR-DGGE, which could be associated with organic or inorganic sulfur cycling. The outcome of the present study highlights ecophysiologically important roles of AnPBs in the organically polluted marine sediments.

山川湾位于日本鹿儿岛县井市,是一个地理上封闭的沿海海洋入口,其不断恶化的海底沉积物处于缺氧、还原性、富硫化物的状态。为了深入了解无氧光合细菌(AnPBs)的多样性及其在沉积物中的生态生理作用,采用了分离AnPBs、16S rDNA PCR-DGGE和pufM PCR-DGGE三种方法。在分离的细菌中,球形红杆菌(Rhodobacter sphaeroides)的近缘菌最为占优势,可能对沉积物中的有机污染物有转化作用。16S rDNA PCR-DGGE分析结果显示,该菌株BS1丰度较高。它可以反映出密集的分层和由此形成的缺氧、富硫化物层,以及该菌株对极弱光的适应。pufM PCR-DGGE还检测到多种紫色无硫或含硫细菌以及好氧无氧光异养生物,这些细菌可能与有机或无机硫循环有关。本研究结果强调了AnPBs在有机污染海洋沉积物中的重要生态生理作用。
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引用次数: 10
期刊
Biocontrol science
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