• Title/Summary/Keyword: culturable bacteria

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Comparison of Metabolic Pathways of Less Orthologous Prokaryotes than Mycoplasma genitalium (Mycoplasma genitalium 보다 보존적 유전자 수가 작은 원핵생물들의 대사경로 비교)

  • Lee, Dong-Geun
    • Journal of Life Science
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    • v.28 no.3
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    • pp.369-375
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    • 2018
  • Mycoplasma genitalium has 367 conserved genes and the smallest genome among mono-culturable prokaryotes. Conservative metabolic pathways were examined among M. genitalium and 14 prokaryotes, one hyperthermophilic exosymbiotic archaeon Nanoarchaeum equitans and 13 intracellular eubacteria of plants or insects, with fewer conserved genes than M. genitalium. They have 11 to 71 metabolic pathways, however complete metabolic pathways ranged from 1 to 24. Totally, metabolic pathway hole is very high due to the lack of 45.8% of the enzymes required for the whole metabolic pathways and it could be suggested that the shared metabolic pathway with the host's enzyme would work or the essential substances are host dependent. The number of genes necessary for mass transfer through the cell membrane is also very low, and it may be considered that the simple diffusion or the protein of the host will function in the cell membrane of these prokaryotes. Although the tRNA charging pathway was distributed in all 15 prokaryotes, each has 5-20 tRNA charging genes. This study would give clues to the understanding of the metabolic pathways of intracellular parasitic bacteria of plant and endosymbiotic bacteria of insects, and could provide basic data for prevention of crop damage, development of insect pests and human medicines.

Diversity of ampicillin resistant bacteria in domestic streams (국내 하천에 분포하는 ampicillin 내성균의 다양성)

  • Go, Eun Byeul;Chung, In-Young;Kim, Hyuk;Seok, Kwang-Seol;Kim, Byeori;Yoo, Yong-Jae;Jang, Yejin;Chae, Jong-Chan
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.440-443
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    • 2015
  • The widespread emergence of antibiotic resistant microorganisms in clinics and natural environments has attracted public concern. Especially, microorganisms inhabiting natural environment is considered as a source responsible for increasing the abundance of antibiotic resistant genes in ecosystem. In this study, the diversity of culturable bacteria resistant to ampicillin was investigated with water samples collected from seven locations in Korea. The genera belonging to Aeromonas and Acidovorax were dominant among the isolated 498 strains. The 66% of isolates showed multi-drug resistance against more than six antibiotics among tested fourteen ones and isolates resistant to seven antibiotics were the most prevalent with 19.7% abundance. Using the antibiotics susceptibility results, the intrinsic resistance profile was suggested for the most dominant genera, Aeromonas, Acidovorax, Pseudomonas, and Elizabethkingia.

Diversity and Phylogenetic Analysis of Culturable Marine Bacteria Isolated from Rhizosphere Soils of Suaeda japonica Makino in Suncheon Bay (순천만 칠면초의 근권으로부터 분리된 해양세균의 다양성 및 계통학적 분석)

  • You, Young-Hyun;Park, Jong Myong;Nam, Yoon-Jong;Kim, Hyun;Lee, Myung-Chul;Kim, Jong-Guk
    • Journal of Life Science
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    • v.25 no.2
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    • pp.189-196
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    • 2015
  • Bacterial diversity was studied in the rhizosphere of Suaeda japonica Makino, which is native to Suncheon Bay in South Korea. Soil samples from several sites were diluted serially, and pure isolation was performed by subculture using marine agar and tryptic soy agar media. Genomic DNA was extracted from 29 pure, isolated bacterial strains, after which their 16S rDNA sequences were amplified and analyzed. Phylogenetic analysis was performed to confirm their genetic relationship. The 29 bacterial strains were classified into five groups: phylum Firmicutes (44.8%), Gamma proteobacteria group (27.6%), Alpha proteobacteria group (10.3%), phylum Bacteriodetes (10.3%), and phylum Actinobacteria (6.8%). The most widely distributed genera were Bacillus (phylum Firmicutes), and Marinobacterium, Halomonas, and Vibrio (Gamma proteobacteria group). To confirm the bacterial diversity in rhizospheres of S. japonica, the diversity index was used at the genus level. The results show that bacterial diversity differed at each of the sampling sites. These 29 bacterial strains are thought to play a major role in material cycling at Suncheon Bay, in overcoming the sea/mud flat-specific environmental stress. Furthermore, some strains are assumed to be involved in a positive interaction with the halophyte S. japonica, as rhizospheric flora, with induction of growth promotion and plant defense mechanism.

Analysis of Microbial Community Structure in Soil and Crop Root System II. Analysis of soil microbial community structure in different soil Environmental conditions by MIDI and DNA analyses (토양과 작물근계의 미생물군집 구조 해석 II. MIDI 및 DNA 분석에 의한 토양환경별 미생물 군집 해석)

  • Ryu, Jin-Chang;Kwon, Soon-Wo;Kim, Jong-Shik;Suh, Jang-Sun;Jung, Beung-Gan;Choi, Sun-Shik
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.2
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    • pp.118-126
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    • 2002
  • To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.

Activity and Survival of the Natural Bacteria under the Stressed Conditions Detected by Bioluminescent Phenotype (스트레스 하의 자연세균의 활성 및 생존의 발광표현형을 이용한 탐지)

  • Park, Kyoung-Je;Yoon, Hye-Young;Chun, Se-Jin;Lee, Ho-Sa;Lee, Dong-Hun;Jahng, Deokjin;Lee, Kyu-Ho
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.154-161
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    • 1998
  • To investigate whether the introduced genetic marker is useful to detect the survivalship and activity of the natural bacteria under the stressed conditions, one Gram-negative isolate, KP964 was transformed to the luminous phenotype by transferring luxAB gene. Under the starvation-stress this luminous bacterial culturability (determined by colony-forming-units [CFU] on agar plate) decreased rapidly below the detection limit by 37 days, while its total cell number (determined by AODC) remained almost the same as its initial inocular size. At that time period, the viable cell number was estimated to be 1400 times higher than its CFU number. The bioiuminescence (determined by relative light units [RLU]) produced under the same condition was also monitored and found to decrease more rapidly than the culturability by 5-fold. Under the other stresses, e.g., osmotic shocks, acid shock, and exposure to toxic chemicals, this bacterial strain did not show the reliable correlation between CFU and RLU. These results might not suggest the direct estimation of bioiuminescence from the stressed bacteria be an index of both the survivalship and its activity. However, when the stressed bacterial cells were incubated under the favorable condition by relieving from the existing stress, the potential bioiuminescence (the lag periods before the increase of bioiuminescence, the increase rates of bioiuminescence, and the maximal levels of bioiuminescence) was shown to be highly dependent upon the strengths of the stresses exposed to the bacterial cells. Therefore, analysis of the potential bioiuminescence from the stressed bacteria revealed good relationships with survival as well as activity.

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Assessment of Inactivation for Campylobacter spp. Attached on Chicken Meat (계육에 오염된 Campylobacter 균의 불활성화 평가)

  • Jang Keum-Il;Jeong Heon-Sang;Kim Chung-Ho;Kim Kwang-Yup
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.302-307
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    • 2005
  • The inactivation efficiency of Campylobacter jejuni were assessed in vitro and in vivo using confocal laser microscopy and flow cytometry. C. jejuni cells were inactivated with $1\%$ (w/v) trisodium phosphate (TSP) and the live cells and inactivated cells were distinguished by staining with LIVE/DEAD BacLight Bacteria Viability fluorescent probe. After treatment of TSP for 5 min, most of C. jejuni cells turned to coccoid form from original spiral shape. C. jejuni cells lost total cell viability in the absence of organic nutrients but did not lost total cell viability in the presence of organic nutrients. In vivo test, C. jejuni cells turned to viable but non-culturable (VBNC) form after TSP treatment and remained alive on chicken skin. C. jejuni cells attached on chicken meat would transform to coccoid form by sanitizer treatment, but could possibly be alive by the benefits of organic nutrients present in chicken meat.

Microbiological Features and Bioactivity of a Fermented Manure Product (Preparation 500) Used in Biodynamic Agriculture

  • Giannattasio, Matteo;Vendramin, Elena;Fornasier, Flavio;Alberghini, Sara;Zanardo, Marina;Stellin, Fabio;Concheri, Giuseppe;Stevanato, Piergiorgio;Ertani, Andrea;Nardi, Serenella;Rizzi, Valeria;Piffanelli, Pietro;Spaccini, Riccardo;Mazzei, Pierluigi;Piccolo, Alessandro;Squartini, Andrea
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.644-651
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    • 2013
  • The fermented manure derivative known as Preparation 500 is traditionally used as a field spray in biodynamic agriculture for maintaining and increasing soil fertility. This work aimed at characterizing the product from a microbiological standpoint and at assaying its bioactive properties. The approach involved molecular taxonomical characterization of the culturable microbial community; ARISA fingerprints of the total bacteria and fungal communities; chemical elemental macronutrient analysis via a combustion analyzer; activity assays for six key enzymes; bioassays for bacterial quorum sensing and chitolipooligosaccharide production; and plant hormone-like activity. The material was found to harbor a bacterial community of $2.38{\times}10^8$ CFU/g dw dominated by Gram-positives with minor instances of Actinobacteria and Gammaproteobacteria. ARISA showed a coherence of bacterial assemblages in different preparation lots of the same year in spite of geographic origin. Enzymatic activities showed elevated values of ${\beta}$-glucosidase, alkaline phosphatase, chitinase, and esterase. The preparation had no quorum sensing-detectable signal, and no rhizobial nod gene-inducing properties, but displayed a strong auxin-like effect on plants. Enzymatic analyses indicated a bioactive potential in the fertility and nutrient cycling contexts. The IAA activity and microbial degradation products qualify for a possible activity as soil biostimulants. Quantitative details and possible modes of action are discussed.

Cloning of α-Amylase Gene from Unculturable Bacterium Using Cow Rumen Metagenome (소 반추위 메타게놈에서 비배양 세균의 α-amylase 유전자 클로닝)

  • Cho, Soo-Jeong;Yun-Han-Dae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1013-1021
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    • 2005
  • The metagenomes of complex microbial communities are rich sources of novel biocatalysts. The gene encoding an extracellular $\alpha$-amylase from a genomic DNA of cow rumen was cloned in Escherichia coli DH5$\alpha$ and sequenced. The $\alpha$-amylase (amyA) gene was 1,893 bp in length, encoding a protein of 631 amino acid residues with calculated molecular weight of 70,734 Da. The molecular weight of the enzyme was estimated to be about 71,000 Da by active staining of a SDS-PACE. The enzyme was 21 to $59\%$ sequence identical with other amyloyltic enzymes. The AmyA was optimally active at pH 6.0 and $40\%$. The AmyA had a calculated pI of 5.87. AmyA expressed in E. coli DH5$\alpha$ was enhanced in the presence of $Mg^{2+}$ (20 mM) and $Ca^{2+}$ (30 mM) and inhibited in the presence of $Fe^{2+}$ and $Cu^{2+}$. The origin of amyA gene could not be confirmed by PCR using internal primer of amyA gene from extracted genomic DNA of 49 species rumen culturable bacteria so far. An amyh is supposed to obtained from unculturable rumen bacterium in cow rumen environment.

Effects of Storage Temperature on the Survival of Vibrio mimicus K-1 in Seawater and Arkshell (해수와 피조개에서 Vibrio mimicus K-1의 생존에 대한 보관온도의 영향)

  • KOH Byeong-Ho;LEE Won-Dong;ANN Sung-Kee;KIM Ji-Hoe;LEE Myung-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.2
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    • pp.277-281
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    • 1997
  • The cell density changes of Vibrio mimicus K-1 in sea water and arkshell feeding it were examined at various temperature. The strain was suspended in sterilized sea water and storaged at experimental temperature $(5,\;10,\;15,\;20,\;and\;28^{\circ}C)$). At intervals of up to 10 days, aliquots of each suspension were plated onto BHI agar. At 5 and $10^{\circ}C$, the plate counts of V. mimicus K-1 showed a rapid decline, which 3s known to be a reault of this bacterium's entering into the viable but non culturable state. At 20 and $28^{\circ}C$, however, V. mimicus K-1 are stable over the 10 days experimental periods. V. mimicus K-1 was fed to arkshell, which was subsequently stored at temperatures ranging from 5 to $20^{\circ}C$ for 10 days. The samples of arkshell were homogenized and plated at intervals to determine the cell density of V. mimicus K-1 and total aerobic population of bacteria present. At 5 and $10^{\circ}C$, the numbers of V. mimicus K-1 in sea water rapid decreased over the 10 days experimental periods. However, little change of V. mimicus K-1 density was observed in shellstock arkshell at 5 and $10^{\circ}C$. While, V. mimicus K-1 density was decreased more rapidly to level below limit of dectection in shucked arkshell at same temperature. Incubation at the higher temperature $(20^{\circ}C)$ resulted in large increase in total aerobic bacterial number of shellstock arkshell. These results suggest that even with proper storage, indigenous levels of V. mimicus may remain sufficiently high in shellstock arkshell to produce infection in compromise hosts.

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