• Title/Summary/Keyword: 16S-rRNA

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Comparison of bacterial communities in leachate from decomposing bovine carcasses

  • Yang, Seung Hak;Ahn, Hee Kwon;Kim, Bong Soo;Chang, Sun Sik;Chung, Ki Yong;Lee, Eun Mi;Ki, Kwang Seok;Kwon, Eung Gi
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1660-1666
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    • 2017
  • Objective: Burial is associated with environmental effects such as the contamination of ground or surface water with biological materials generated during the decomposition process. Therefore, bacterial communities in leachates originating from the decomposing bovine carcasses were investigated. Methods: To understand the process of bovine (Hanwoo) carcass decomposition, we simulated burial using a lab-scale reactor with a volume of $5.15m^3$. Leachate samples from 3 carcasses were collected using a peristaltic pump once a month for a period of 5 months, and bacterial communities in samples were identified by pyrosequencing of the 16S rRNA gene. Results: We obtained a total of 110,442 reads from the triplicate samples of various sampling time points (total of 15 samples), and found that the phylum Firmicutes was dominant at most sampling times. Differences in the bacterial communities at the various time points were observed among the triplicate samples. The bacterial communities sampled at 4 months showed the most different compositions. The genera Pseudomonas and Psychrobacter in the phylum Proteobacteria were dominant in all of the samples obtained after 3 months. Bacillaceae, Clostridium, and Clostridiales were found to be predominant after 4 months in the leachate from one carcass, whereas Planococcaceae was found to be a dominant in samples obtained at the first and second months from the other two carcasses. The results showed that potentially pathogenic microbes such as Clostridium derived from bovine leachate could dominate the soil environment of a burial site. Conclusion: Our results indicated that the composition of bacterial communities in leachates of a decomposing bovine shifted continuously during the experimental period, with significant changes detected after 4 months of burial.

Microbial Floral Dynamics of Chinese Traditional Soybean Paste (Doujiang) and Commercial Soybean Paste

  • Gao, Xiuzhi;Liu, Hui;Yi, Xinxin;Liu, Yiqian;Wang, Xiaodong;Xu, Wensheng;Tong, Qigen;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1717-1725
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    • 2013
  • Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

Bacillus oryzicola sp. nov., an Endophytic Bacterium Isolated from the Roots of Rice with Antimicrobial, Plant Growth Promoting, and Systemic Resistance Inducing Activities in Rice

  • Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.152-164
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    • 2015
  • Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.

Ecological Characteristics of Actinomycetes from Mercury and Chrome Polluted Soil (수은, 6가크롬 오염토양으로부터 분리된 방선균군의 생태학적 특성)

  • Cho Min-Hye;Han Sang-Mi;Baek Ha-Ju;Whang Kyung-Sook
    • Korean Journal of Environmental Biology
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    • v.24 no.1 s.61
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    • pp.38-45
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    • 2006
  • Ecological characteristics of microbial populations inhabiting heavy metal polluted soil were investigated. The samples were collected from 293 sites around an factory and industry at Gyeoungsangbuk-do. We measured the contents of seven heavy metal elements (Cd, Cu, As, Hg, Pb, $Cr^{6+}$, CN), seven sites have been seriously contaminated by mercury and chrome. A quantitative evaluation of microbial populations in mercury and chrome contaminated soil was examined by using plate count method. Bacterial numbers in polluted soil samples ranged from $7.4X10^5\;to\;9.3X10^7\;cfu\;g^{-1}$, about $10\sim100$ fold less than the count for the unpolluted soil. Moulds were not detected in chrome polluted soil. The log values of actinomycetes of each contaminated soil samples were log ranged from 6.18 to 7.52. The ratio of actinomycetes was similar to unpolluted soil. The investigation showed actinomycetes to be the major microbial population inhabiting the mercury and chrome polluted soil. Thirty-one isolates among the total isolates were examined for antibacterial activity. These isolates were identified based on a phylogenetic analysis using 16S rRNA gene nucleotide sequences, they were categorized in three major phylogenetic groups, belong to the Streptomyces (6 strains), Saccharopolyspora (3 strains), Nocardiodes (1 strain). On the phylogenetic tree, the clade consisting of five isolates were distantly related to all of the established Streptomycetes genera, indicating the possibility as members of new species.

Selection of Biogenic Amine-reducing Microorganisms from a Traditional Korean-style Fermented Food, $Cheonggukjang$ (전통 발효 청국장으로부터 biogenic amine 저생성 미생물의 선발)

  • Choi, Jae-Young;Hong, Sung-Wook;Chung, Kun-Sub
    • Korean Journal of Food Science and Technology
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    • v.44 no.2
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    • pp.196-201
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    • 2012
  • Microorganisms, having the lower decarboxylase activity, among the isolated strains from $cheonggukjang$ and rice-straw in this study were selected by using biogenic amine (BA) media. The selected strains were identified as $Bacillus$ $subtilis$ HH12, $B.$ $subtilis$ HR254, and $Paenibacillus$ $barcinonensis$ KR97, by using 16S rRNA analysis. PCR analysis showed that the histidine decarboxylase ($hdc$) gene was absent in the HH12, HR254, and KR97 strains. However, PCR analysis showed that the tyrosine decarboxylase ($tdc$) gene was present in the HH12, HR254, and KR97 strains. Quantitative analysis of the selected strains by using high-performance liquid chromatography showed that histamine was absent in the HH12, HR254, and KR97 strains. However, these 3 strains showed tyramine concentrations of 6.09, 3.68, and 6.30 mg/L, respectively. These strains produced lower concentrations of amines (approximately 7.9, 0, and 9.3% amines in the HH12, HR254, and KR97 strains, respectively) than the $B.$ $subtilis$ MC138 strain, which showed the higher protease activity.

Multiple Confirmation and RAPD-genotyping of Enterobacter sakazakii Isolated from Sunsik (선식에서 분리한 Enterobacter sakazakii의 복합동정 및 RAPD를 이용한 genotyping)

  • Choi, Jae-Won;Kim, Yun-Ji;Lee, Jong-Kyung;Kim, Young-Ho;Kwon, Ki-Sung;Hwang, In-Gyun;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.40 no.1
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    • pp.101-105
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    • 2008
  • Enterobacter sakazakii is implicated in severe forms of neonatal infections such as meningitis and sepsis. This organism has been isolated from a wide range of foods, including cheese, vegetables, grains, herbs, and spices, but its primary environment is still unknown. Generally, dried infant milk formula has been epidemiologically identified as the source of E. sakazakii. Sunsik (a powdered mixture of roasted grains and other foodstuffs) is widely consumed in Korea as a side dish or energy supplement. Sunsik is consumed without heat treatment; thus, lacking an additional opportunity to inactivate foodborne pathogens. Therefore, its microbiological safety should be guaranteed. In this study, the prevalence of E. sakazakii was monitored in 23 different sunsik component flours, using FDA recommended methods; but E. sakazakii medium (Neogen) and Chromogenic E. sakazakii medium (Oxoid) were used as the selective media. In total, presumptive E. sakazakii strains were isolated from 8 different sunsik powders. Subsequently, an API 20E test was conducted, and 15 strains from 5 different sunsik flours (sea tangle, brown rice, non-glutinous rice, cheonggukjang, dried anchovy) were confirmed as E. sakazakii. Fifteen strains were again confirmed by PCR amplification, using three different primer sets (tDNA sequence, ITS sequence, 16S rRNA sequence), and compared to ATCC strains (12868, 29004, 29544, 51329). They were once again confirmed by their enzyme production profiles using an API ZYM kit. Finally, RAPD (random amplified polymorphic DNA)-genotyping was carried out as a monitoring tool to determine the contamination route of E. sakazakii during processing.

Development of a new lactic acid bacterial inoculant for fresh rice straw silage

  • Kim, Jong Geun;Ham, Jun Sang;Li, Yu Wei;Park, Hyung Soo;Huh, Chul-Sung;Park, Byung-Chul
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.950-956
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    • 2017
  • Objective: Effects of newly isolated Lactobacillus plantarum on the fermentation and chemical composition of fresh rice straw silage was evaluated in this study. Methods: Lactic acid bacteria (LAB) from good crop silage were screened by growing them in MRS broth and a minimal medium with low carbohydrate content. Selected LAB (LAB 1821) were Gram-positive, rods, catalase negative, and were identified to be Lactobacillus plantarum based on their biochemical characteristics and a 16S rRNA analysis. Fresh rice straw was ensiled with two isolated LAB (1821 and 1841), two commercial inoculants (HM/F and P1132) and no additive as a control. Results: After 2 months of storage at ambient temperature, rice straw silages treated with additives were well-preserved, the pH values and butyric and acetic acid contents were lower, and the lactic acid content and lactic/acetic acid ratio were higher than those in the control (p<0.05). Acidity (pH) was lowest, and lactic acid highest, in 1821-treated silage (p<0.05). The $NH_3-N$ content decreased significantly in inoculant-treated silage (p<0.05) and the $NH_3-N$ content in 1821-treated silage was lowest among the treatments. The dry matter (DM) content of the control silage was lower than that of fresh rice straw (p<0.05), while that of the 1841- and p1174-inoculant-treated silages was significantly higher than that of HM/F-treated silage. Microbial additives did not have any significant (p>0.05) effect on acid detergent fiber or neutral detergent fiber contents. Crude protein (CP) content and in vitro DM digestibility (IVDMD) increased after inoculation of LAB 1821 (p<0.05). Conclusion: LAB 1821 increased the CP, IVDMD, lactic acid content and ratio of lactic acid to acetic acid in rice straw silage and decreased the pH, acetic acid, $NH_3-N$, and butyric acid contents. Therefore, adding LAB 1821 improved the fermentation quality and feed value of rice straw silage.

Isolation of Bacillus velezensis SSH100-10 with Antifungal Activity from Korean Traditional Soysauce and Characterization of Its Antifungal Compounds (전통재래 간장으로부터 항진균 활성 B. velezensis SSH100-10의 분리와 그 항진균 물질의 특성 구명)

  • Chang, Mi;Moon, Song Hee;Chang, Hae Choon
    • Food Science and Preservation
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    • v.19 no.5
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    • pp.757-766
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    • 2012
  • The SSH100-10 bacterial strain, which exhibits strong antifungal (anti-mold and anti-yeast) activity, was isolated from traditional korean soysauce aged 100 years. The strain was identified as Bacillus velezensis based on Gram-staining, the biochemical properties and 16S rRNA gene sequence determination. B. velezensis SSH100-10 showed strong proteinase activity and NaCl tolerance, but did not produce enterotoxin. Two-antifungal compounds from B. velezensis SSH100-10 were purified using SPE, preparative HPLC, and reverse phase-HPLC. The purified antifungal compounds were identified as $C_{14}$ and $C_{15}$ iturin through MALDI-TOF-MS and amino acid composition analysis. The stability characteristics of the antifungal compounds after temperature, pH, and enzyme treatments suggested that B. velezensis SSH100-10 produced more than two antifungal compounds; pH-stable $C_{14}$ iturin A and $C_{15}$ iturin A, and unidentified pH-unstable compounds. The results suggested that B. velezensis SSH100-10 can be used in soybean fermentation as a starter. Moreover it has potential as a biopreservative in the food and feed industry and as a biocontrol agent in the field of agriculture.

In vitro Antimicrobial Activity of a New Isolate Streptomyces sp. BCNU 1030 (신규 분리균주 Streptomyces sp. BCNU 1030의 in vitro 항균활성)

  • Bang, Ji-Hun;Choi, Hye-Jung;Ahn, Cheol-Soo;Kim, Dong-Wan;Jeong, Yong-Kee;Joo, Woo-Hong
    • Journal of Life Science
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    • v.21 no.4
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    • pp.589-595
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    • 2011
  • This work focused on screening and characterizing antibiotic-producing actinomycetes to develop new antibiotics that can overcome the growing resistance of disease-causing microbes. One-hundred actinomycetes strains were isolated from soil samples from Chungcheongbuk-do, Korea using various kinds of actinomycetes isolation media, including a starch casein agar medium and potato dextrose agar (PDA). Among them, strain BCNU 1030 was determined to show strong antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA). Biochemical, physiological, and 16S rRNA sequence analyses indicated that strain BCNU 1030 belonged to the genus Streptomyces. Strain BCNU 1030 exhibited antibiotic activity against a wide range of bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration (MIC) of BCNU 1030 dichloromethane extract was determined to be $0.78\;{\mu}g/ml$ for MRSA CCARM 3090. Therefore, Streptomyces sp. BCNU 1030 has potential for anti-MRSA drug development.

Characteristic study and isolation of Bacillus subtilis SRCM 101269 for application of cow manure (우분 적용을 위한 Bacillus subtilis SRCM 101269의 분리 및 특성 연구)

  • Jeon, SaeBom;Oh, HyeonHwa;Uhm, Tai-Boong;Cho, Jae-Young;Yang, Hee-Jong;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.74-83
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    • 2016
  • Bacillus subtilis SRCM 101269 having safety and amo gene isolated from Korean traditional fermented food and their investigated characterization to apply the cow manure such as cellulase and xylanase activities, 16S rRNA sequencing, and ability of removal of livestock manure odor. Cow manure application results for the removal of livestock manure odor, the ammonia gas was reduced more than two-folder compared to the control group after 6 days, and reduced to less than 10 ppm after 9 days. In the case of cow manure added fowl droppings and other wood-based mixture components, ammonia gas maintained constant after 3 days of fermentation. However, in the case of sample inoculated B. subtilis SRCM 101269, ammonia gas reduced in course of fermentation time, and concentration of hydrogen sulfide also reduced for 65 ppm. Changes of nitrite concentration according to fermentation time no showed different for cow manure, however nitrite concentration in mixed livestock manure increased when compared to control. And then sulfate concentration in cow manure decreased, and no showed different when compared to the initial fermentation. No apparent change of sulfate concentration in mixed livestock manure detected. Through the previously studies, B. subtilis SRCM 101269 has high potential in industrial application manufacturing the cow manure as removal of livestock manure odor.