• Title/Summary/Keyword: 16S-23S rRNA

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Short-Term Effect of Elevated Temperature on the Abundance and Diversity of Bacterial and Archaeal amoA Genes in Antarctic Soils

  • Han, Jiwon;Jung, Jaejoon;Park, Minsuk;Hyun, Seunghun;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • 제23권9호
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    • pp.1187-1196
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    • 2013
  • Global warming will have far-reaching effects on our ecosystem. However, its effects on Antarctic soils have been poorly explored. To assess the effects of warming on microbial abundance and community composition, we sampled Antarctic soils from the King George Island in the Antarctic Peninsula and incubated these soils at elevated temperatures of $5^{\circ}C$ and $8^{\circ}C$ for 14 days. The reduction in total organic carbon and increase in soil respiration were attributed to the increased proliferation of Bacteria, Fungi, and Archaea. Interestingly, bacterial ammonia monooxygenase (amoA) genes were predominant over archaeal amoA, unlike in many other environments reported previously. Phylogenetic analyses of bacterial and archaeal amoA communities via clone libraries revealed that the diversity of amoA genes in Antarctic ammonia-oxidizing prokaryotic communities were temperature-insensitive. Interestingly, our data also showed that the amoA of Antarctic ammonia-oxidizing bacteria (AOB) communities differed from previously described amoA sequences of cultured isolates and clone library sequences, suggesting the presence of novel Antarctic-specific AOB communities. Denitrification-related genes were significantly reduced under warming conditions, whereas the abundance of amoA and nifH increased. Barcoded pyrosequencing of the bacterial 16S rRNA gene revealed that Proteobacteria, Acidobacteria, and Actinobacteria were the major phyla in Antarctic soils and the effect of short-term warming on the bacterial community was not apparent.

Isolation of an Indigenous Imidacloprid-Degrading Bacterium and Imidacloprid Bioremediation Under Simulated In Situ and Ex Situ Conditions

  • Hu, Guiping;Zhao, Yan;Liu, Bo;Song, Fengqing;You, Minsheng
    • Journal of Microbiology and Biotechnology
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    • 제23권11호
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    • pp.1617-1626
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    • 2013
  • The Bacterial community structure and its complexity of the enrichment culture during the isolation and screening of imidacloprid-degrading strain were studied using denaturating gradient gel electrophoresis analysis. The dominant bacteria in the original tea rhizosphere soil were uncultured bacteria, Rhizobium sp., Sinorhizobium, Ochrobactrum sp., Alcaligenes, Bacillus sp., Bacterium, Klebsiella sp., and Ensifer adhaerens. The bacterial community structure was altered extensively and its complexity reduced during the enrichment process, and four culturable bacteria, Ochrobactrum sp., Rhizobium sp., Geobacillus stearothermophilus, and Alcaligenes faecalis, remained in the final enrichment. Only one indigenous strain, BCL-1, with imidacloprid-degrading potential, was isolated from the sixth enrichment culture. This isolate was a gram-negative rod-shaped bacterium and identified as the genus Ochrobactrum based on its morphological, physiological, and biochemical properties and its 16S rRNA gene sequence. The degradation test showed that approximately 67.67% of the imidacloprid (50 mg/l) was degraded within 48 h by strain BCL-1. The optimum conditions for degradation were a pH of 8 and $30^{\circ}C$. The simulation of imidacloprid bioremediation by strain BCL-1 in soil demonstrated that the best performance in situ (tea soil) resulted in the degradation of 92.44% of the imidacloprid (100 mg/g) within 20 days, which was better than those observed in the ex situ simulations that were 64.66% (cabbage soil), 41.15% (potato soil), and 54.15% (tomato soil).

Report of 21 unrecorded bacterial species in Korea belonging to Betaproteobacteria and Epsilonproteobacteria

  • Kim, Min-Kyeong;Seong, Chi-Nam;Jahng, Kwangyeop;Cha, Chang-Jun;Joh, Ki-seong;Bae, Jin-Woo;Cho, Jang-Cheon;Im, Wan-Taek;Kim, Seung-Bum
    • Journal of Species Research
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    • 제6권1호
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    • pp.15-24
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    • 2017
  • During the extensive survey of the prokaryotic species diversity in Korea, bacterial strains belonging to Betaproteobacteria and Epsilonproteobacteria were isolated from various sources including freshwater, sediment, soil and fish. A total of 23 isolates were obtained, among which 22 strains were assigned to the class Betaproteobacteria and one strain to the class Epsilonproteobacteria. The 22 betaproteobacterial strains were further assigned to Comamonadaceae (11 strains), Burkholderiaceae (6 strains), Oxalobacteraceae (2 strains), Neisseriaceae (1 strain) and unclassified family groups (2 strains). For the strains of Burkholderiaceae, 3 strains were identified as 3 species of Burkholderia, and 2 strains were as 2 species of Cupriavidus. For the strains of Comamonadaceae, 4 strains were identified as 2 species of the genus Hydrogenophaga, 2 strains as 2 species of Acidovorax, 2 strains as 2 species of Limnohabitans, and each of the remaining strains as single species of Comamonas, Curvibacter and Rhodoferax, respectively. For the strains of Oxalobacteraceae, 1 strain was identified as a species of Undibacterium, and the other strain as a species of Herbaspirillum. The strain belonging to Neisseriaceae was identified as a species of Iodobacter. The remaining strains of Betaproteobacteria were identified as species of Sphaerotilus and Methylibium respectively (family unassigned). The epsilonproteobacterial strain was identified as a species of Arcobacter of the family Camplyobacteraceae. The detailed description of each unrecorded species is provided.

Cyanobacterial Diversity Shifts Induced by Butachlor in Selected Indian Rice Fields in Eastern Uttar Pradesh and Western Bihar Analyzed with PCR and DGGE

  • Kumari, Nidhi;Narayan, Om Prakash;Rai, Lal Chand
    • Journal of Microbiology and Biotechnology
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    • 제22권1호
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    • pp.1-12
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    • 2012
  • The present study examines the effects of 30 mg/kg butachlor on the cyanobacterial diversity of rice fields in Eastern Uttar Pradesh and Western Bihar in India. A total of 40 samples were grouped into three classes [(i) acidic, (ii) neutral, and (iii) alkaline soils], based on physicochemical and principle component analyses. Acidic soils mainly harbored Westillopsis, Trichormus, Anabaenopsis, and unicellular cyanobacteria; whereas Nostoc, Anabaena, Calothrix, Tolypothrix, and Aulosira were found in neutral and alkaline soils. Molecular characterization using 16S rRNA PCR and DGGE revealed the presence of 13 different phylotypes of cyanobacteria in these samples. Butachlor treatment of the soil samples led to the disappearance of 5 and the emergence of 2 additional phylotypes. A total of 40 DGGE bands showed significant reproducible changes upon treatment with butachlor. Phylogenetic analyses divided the phylotypes into five major clusters exhibiting interesting links with soil pH. Aulosira, Anabaena, Trichormus, and Anabaenopsis were sensitive to butachlor treatment, whereas uncultured cyanobacteria, a chroococcalean member, Westillopsis, Nostoc, Calothrix, Tolypothrix, Rivularia, Gloeotrichia, Fischerella, Leptolyngbya, and Cylindrospermum, appeared to be tolerant against butachlor at their native soil pH. Butachlor-induced inhibition of nitrogen fixation was found to be 65% (maximum) and 33% (minimum) in the soil samples of pH 9.23 and 5.20, respectively. In conclusion, low butachlor doses may prove beneficial in paddy fields having a neutral to alkaline soil pH.

Conjugated linoleic acid producing potential of lactobacilli isolated from goat (AXB) rumen fluid samples

  • Tyagi, Amrish Kumar;Kumar, Sachin;Choudhury, Prasanta Kumar;Tyagi, Bhawna;Tyagi, Nitin
    • Asian-Australasian Journal of Animal Sciences
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    • 제33권8호
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    • pp.1233-1241
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    • 2020
  • Objective: The present investigation was aimed to explore the potential of lactobacilli for conjugated linoleic acid (CLA) production, isolated from rumen fluid samples of lactating goats. Methods: A total of 64 isolates of lactobacilli were obtained using deMan-Rogosa-Sharpe (MRS) agar from rumen fluid of goats and further subjected to morphological and biochemical characterizations. Isolates found as gram-positive, catalase negative rods were presumptively identified as Lactobacillus species and further confirmed by genus specific polymerase chain reaction (PCR). The phylogenetic tree was constructed from the nucleotide sequences using MEGA6. Results: Out of the 64 isolates, 23 isolates were observed positive for CLA production by linoleate isomerase gene-based amplification and quantitatively by UV-spectrophotometric assay for the conversion of linoleic acid to CLA as well as gas chromatography-based assay. In all Lactobacillus species cis9, trans11 isomer was observed as the most predominant CLA isomer. These positive isolates were identified by 16S rRNA gene-based PCR sequencing and identified to be different species of L. ingluviei (2), L.salivarius (2), L. curvatus (15), and L. sakei (4). Conclusion: The findings of the present study concluded that lactic acid bacteria isolated from ruminal fluid samples of goat have the potential to produce bioactive CLA and may be applied as a direct fed microbial to enhance the nutraceutical value of animal food products.

Mitigating $CH_4$ Emissions in Semi-Aerobic Landfills: Impacts of Operating Conditions on Abundance and Community Structure of Methanotrophs in Cover Soils

  • Li, Huai;Chi, Zi-Fang;Lu, Wen-Jing;Wang, Hong-Tao
    • Journal of Microbiology and Biotechnology
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    • 제23권7호
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    • pp.993-1003
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    • 2013
  • Methanotrophs are the most important sink of $CH_4$, which is a more highly potent greenhouse gas than $CO_2$. Methanotrophic abundance and community diversity in cover soils from two typical semi-aerobic landfills (SALs) in China were detected using real-time polymerase chain reaction (real-time-PCR) and denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes, respectively. Real time-PCR showed that Type I methanotrophs ranged from $1.07{\times}10^6$ to $2.34{\times}10^7$ copies/g soil and that of Type II methanotrophs from $1.51{\times}10^7$ to $1.83{\times}10^8$ copies/g soil. The ratio of Type II to Type I methanotrophic copy numbers ranged from 5.61 to 21.89, indicating that Type II methanotrophs dominated in SAL. DGGE revealed that Type I methanotrophs responded more sensitively to the environment, changing as the community structure varied with different soil types and locations. Methylobacter, Methylosarcina, and Methylomicrobium for Type I, and Methylocystis for Type II were most prevalent in the SAL cover layer. Abundant interflow $O_2$ with high $CH_4$ concentration in SALs is the reason for the higher population density of methanotrophs and the higher enrichment of Type II methanotrophs compared with anaerobic landfills and other ecosystems, which proved a conclusion that increasing the oxygen supply in a landfill cover layer would greatly improve $CH_4$ mitigation.

장기간 발효 김치인 묵은지에서 분리한 Lactobacillus sakei JK-17의 기능성 조사 (Functional Characterization of Lactobacillus sakei JK-17 Isolated from Long-term Fermented Kimchi, Muk Eun Ji)

  • 김동선;조형우;김대한;오계헌
    • KSBB Journal
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    • 제28권1호
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    • pp.18-23
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    • 2013
  • The purpose of this work was to investigate the several functional characteristics of Lactobacillus sakei JK-17 isolated from long-term fermented kimchi, Muk Eun Ji. Initially, phylogenetic analysis using 16S rRNA sequencing was performed to identify the isolate JK-17, and the strain could be assigned to Lactobacillus sakei and designated as L. sakei JK-17. The strain was registered in GenBank as [JX841311]. The changes of bacterial growth and residual organic acids were monitored and HPLC was used to measure quantitatively two organic acids, lactic acid and acetic acid, produced in the culture during 84 hours of incubation. During the incubation period, several functional characteristics of L. sakei JK-17 were examined. L. sakei JK-17 culture depleted nitrite concentration 94.75%. Antioxidant activity of cultural supernatants of L. sakei JK-17 was approx. 53.8%, and ${\beta}$-galactosidase activities were 0.243 units/mL at pH 7.0 and 0.387 units/mL at pH 4.1, respectively. The antibacterial activities against food-poisoning causing bacteria were examined with 20-fold concentrated culture supernatants from L. sakei JK-17 and the antibacterial effects were clearly observed against all bacteria tested in this work.

우리나라 연안해역 난 및 자치어의 분포특성 (Distribution of Eggs and Larvae in Coastal Waters of Korea)

  • 백정익;지환성;유효재;황강석;김두남
    • 한국수산과학회지
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    • 제54권4호
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    • pp.467-479
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    • 2021
  • The distribution and abundance of fish eggs and larvae were investigated from February to December 2020 along the coastal waters of Korea. The eggs and larvae were identified using the mitochondrial DNA cytochrome c oxidase subunit I (mtDNA COI) and 16s rRNA gene. During the study period, eggs of overall 45 taxa belonging to 26 families were collected and larvae of overall 39 taxa belonging to 23 families were collected. In Yeongil Bay, eggs of Engraulis japonicus, which accounted for 83.9% of the total population, was the most dominant species, followed by Sardinops sagax (4.0%), Repomucenus valenciennei (3.8%) and E. japonicus larvae, which accounted for 34.9% of the total population. These were followed by Sebastiscus marmoratus (31.0%). In Gomso Bay, E. japonicus eggs accounted for 61.7% of the total population, followed by Sillago japonica (14.0%), Johnius grypotus (8.8%) and Pholis fangi larvae, which accounted for 53.5% of the total population, followed by Ammodytes personatus (34.1%). In Jinhae Bay, E. japonicus eggs accounted for 86.0% of the total population, followed by Leiognathus nuchalis (4.1%), Konosirus punctatus (3.7%) and E. japonicus larvae, which accounted for 48.7% of the total population, followed by Parablennius yatabei (21.6%).

A comparative study of the prevalence of Helicobacter pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka

  • Mallikaarachchi, MADKS;Rajapakse, Sanath;Gunawardhana, KSND;Jayatilake, JAMS
    • International Journal of Oral Biology
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    • 제46권1호
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    • pp.60-65
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    • 2021
  • Dental health care workers (DHCW) are at a risk of occupational exposure to Helicobacter pylori from the aerosolized oral biofilms and saliva of patients. We designed this study to investigate the prevalence of H. pylori in the oral biofilms of a group of dental and non-dental undergraduates from Sri Lanka. After obtaining informed consent, oral biofilms were collected from 38 dental undergraduates (19 males and 19 females) undergoing clinical training and 33 non-dental undergraduates (14 males and 19 females). The participants were in the age range of 22-27 years and had healthy periodontium. Total DNA from the oral biofilms were extracted, and H. pylori DNA was detected using polymerase chain reaction (PCR) amplification of 16S rRNA gene of H. pylori using JW22-JW23 primers, and the results were confirmed using PCR amplification of H. pylori-urease specific HPU1-HPU2 primers. Out of 71 participants, 11 (28.95%) dental and 3 (9.09%) non-dental undergraduates had H. pylori in their oral biofilms indicating an overall prevalence rate of 19.72% (14/71). Thus, the prevalence of H. pylori in oral biofilms was significantly higher in dental undergraduates than in non-dental undergraduates (p < 0.05). An odds ratio of 4.07 indicated that dental undergraduates were four times more likely to harbor H. pylori in their oral biofilms than non-dental undergraduates. Foregoing data support the fact that there may be greater occupational risk of exposure to H. pylori for dental undergraduates during clinical training than that for non-dental undergraduates, warranting meticulous infection control practices during clinical dentistry.

Evaluation of host and bacterial gene modulation during Lawsonia intracellularis infection in immunocompetent C57BL/6 mouse model

  • Kirthika, Perumalraja;Park, Sungwoo;Jawalagatti, Vijayakumar;Lee, John Hwa
    • Journal of Veterinary Science
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    • 제23권3호
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    • pp.41.1-41.15
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    • 2022
  • Background: Proliferative enteritis caused by Lawsonia intracellularis undermines the economic stability of the swine industry worldwide. The development of cost-effective animal models to study the pathophysiology of the disease will help develop strategies to counter this bacterium. Objectives: This study focused on establishing a model of gastrointestinal (GI) infection of L. intracellularis in C57BL/6 mice to evaluate the disease progression and lesions of proliferative enteropathy (PE) in murine GI tissue. Methods: We assessed the murine mucosal and cell-mediated immune responses generated in response to inoculation with L. intracellularis. Results: The mice developed characteristic lesions of the disease and shed L. intracellularis in the feces following oral inoculation with 5 × 107 bacteria. An increase in L. intracellularis 16s rRNA and groEL copies in the intestine of infected mice indicated intestinal dissemination of the bacteria. The C57BL/6 mice appeared capable of modulating humoral and cell-mediated immune responses to L. intracellularis infection. Notably, the expression of genes for the vitamin B12 receptor and for secreted and membrane-bound mucins were downregulated in L. intracellularis -infected mice. Furthermore, L. intracellularis colonization of the mouse intestine was confirmed by the immunohistochemistry and western blot analyses. Conclusions: This is the first study demonstrating the contributions of bacterial chaperonin and host nutrient genes to PE using an immunocompetent mouse model. This mouse infection model may serve as a platform from which to study L. intracellularis infection and develop potential vaccination and therapeutic strategies to treat PE.