• Title/Summary/Keyword: oligotrophic media

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Oligotrophic Media Compared with a Tryptic Soy Agar or Broth for the Recovery of Burkholderia cepacia Complex from Different Storage Temperatures and Culture Conditions

  • Ahn, Youngbeom;Lee, Un Jung;Lee, Yong-Jin;LiPuma, John J.;Hussong, David;Marasa, Bernard;Cerniglia, Carl E.
    • Journal of Microbiology and Biotechnology
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    • v.29 no.10
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    • pp.1495-1505
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    • 2019
  • The Burkholderia cepacia complex (BCC) is capable of remaining viable in low-nutrient environments and harsh conditions, posing a contamination risk in non-sterile pharmaceutical products as well as a challenge for detection. To develop optimal recovery methods to detect BCC, three oligotrophic media were evaluated and compared with nutrient media for the recovery of BCC from autoclaved distilled water or antiseptic solutions. Serial dilutions ($10^{-1}$ to $10^{-12}CFU/ml$) of 20 BCC strains were inoculated into autoclaved distilled water and stored at $6^{\circ}C$, $23^{\circ}C$ and $42^{\circ}C$ for 42 days. Six suspensions of Burkholderia cenocepacia were used to inoculate aqueous solutions containing $5{\mu}g/ml$ and $50{\mu}g/ml$ chlorhexidine gluconate (CHX) and $10{\mu}g/ml$ benzalkonium chloride (BZK), and stored at $23^{\circ}C$ for a further 199 days. Nutrient media such as Tryptic Soy Agar (TSA) or Tryptic Soy Broth (TSB), oligotrophic media (1/10 strength TSA or TSB, Reasoner's $2^{nd}$ Agar [R2A] or Reasoner's $2^{nd}$ Broth [R2AB], and 1/3 strength R2A or R2AB) were compared by inoculating these media with BCC from autoclaved distilled water and from antiseptic samples. The recovery of BCC in water or antiseptics was higher in culture broth than on solid media. Oligotrophic medium showed a higher recovery efficiency than TSA or TSB for the detection of 20 BCC samples. Results from multiple comparisons allowed us to directly identify significant differences between TSA or TSB and oligotrophic media. An oligotrophic medium pre-enrichment resuscitation step is offered for the United States Pharmacopeia (USP) proposed compendial test method for BCC detection.

Distribution of Oligotrophic Actinomycetes in Forest Soil. (산림 토양 중 저 영양성 방선균의 분포)

  • Jeong, Eun-Ye;Lee, Sang-Hwa;Park, Dong-Jin;Lee, Hyang-Burm;Piao, Zhe;Kim, Chang-Jin
    • Microbiology and Biotechnology Letters
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    • v.28 no.4
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    • pp.239-241
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    • 2000
  • From 5 forest soil samples, the isolation frequency of oligotrophic Actinomycetes was investigated with nutrient and minimal media. The frequency of soil Actinomycetes by the minimal media. The frequency of soil Actinomycetes by the minimal media of DHV, WA, BA and NA was similar to the value by HV nutrient medium. In addition, different Actinomycetes were isolated from all the media used in this experiment and then their growth were tested on nutrient (Bennett's agar) and minimal media(WA). In consequent, the number of different Actinomycetes from minimal media was 26.1 strains, whereas the number of from nutrient medium (HV) was 26.6 strains. Furthermore, the percentage of facultative and obligate strains among the oligotrophic Actinomycetes was 90% and 10%, respectively.

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Isolation and Characterization of Oligotrophic Bacteria Possessing Induced Systemic Disease Resistance against Plant Pathogens

  • Han, Song-Hee;Kang, Beom-Ryong;Lee, Jang-Hoon;Kim, Hyun-Jung;Park, Ju-Yeon;Kim, Jeong-Jun;Kim, Young-Cheol
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.68-74
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    • 2012
  • Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

Evaluation of Various Oligotrophic Media for Cultivation of Previously Uncultured Soil Bacteria (난배양성 토양세균의 배양법 평가 및 신 분류군의 순수분리)

  • Kim, Do-Hyoung;Lee, Sang-Hoon;Cho, Jae-Chang
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.352-357
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    • 2008
  • We evaluated cultivation methods to obtain pure cultures of previously uncultivated bacteria from soil. Soil bacteria (suspensions) were inoculated onto various oligotrophic media with one of the following additives: 1) soil extract; 2) anthraquinone disulfonate (humic acid analogue); 3) acyl homoserine lactones (quorum-signaling compounds); 4) catalase (for the protection of bacteria from exogenous peroxides). After the relatively long period (60 days) of incubation with elevated concentrations of $CO_2$ (5%, v/v), the media containing catalase showed the highest colony count. We purified 147 randomly selected colonies from the media and the isolates were subjected to the phylogenetic analyses of their 16S rRNA gene sequences. Phylogenetic analysis revealed that approximately 30% of the isolates might belong to novel species or novel family, suggesting that the media and incubation conditions used could be useful for the cultivation of as-yet-uncultured bacteria. Especially, bacteria belonging to the phylum Acidobacteria, ubiquitous bacterial taxon known as an uncultured bacterial group (at least difficult to culture from environmental samples), were successfully cultured in this study.

Some Aspects to the in vivo Nitrate Reductase Activity in Carex species (사초속 식물의 질산환원효소 활성의 특징)

  • 추연식
    • Journal of Life Science
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    • v.10 no.1
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    • pp.52-60
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    • 2000
  • Up to now, there have been done much efforts in regard to nitrate reductase activity (NRA) of dicotyledonous herbs and important crop monocotyledons, but few to wild plants having canopy structure such as Carex. The objective of the present study are to determine: a) the optimum in vivo NR assay conditions for leaf samples of Carex species, b) changes of NRA according to section within leaf and leaf ages, c) diurnal variations. Optimized assay media of each Carex species were determined. NRA of C. rostrata adapted to oligotrophic habitats is readily saturated at lower substrate concentration than those of C. distans and C. gracilis, adapted to meso- and eutrophic habitats, respectively. All Carex species investigated have higher NRA in leaves than in roots. NRA of all species showed maximal values at the middle section of each leaf and in the youngest fully expanded leaves. Compared to C. gracilis, NR in leaves of C. distans was adapted readily to the light period. On the whole, Carex showed rather delayed diurnal variation. Even if the in vivo nitrate reductase assay based on nitrite estimation does not give an accurate estimation of total nitrate reduced, it still serves as a useful tool to find out relative differences in varying environmental conditions. Additionally, in vivo RNA measurements are helpful to understand nitrate reduction and basic nitrogen metabolism of Carex species having different canopy structure.

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Arsenic Adsorption onto Pseudomonas aeruginosa Cell Surface (Pseudomonas aeruginosa 표면에 대한 비소의 흡착특성)

  • Lee Jong-Un;Park Hyun-Sung
    • Economic and Environmental Geology
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    • v.38 no.5 s.174
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    • pp.525-534
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    • 2005
  • Adsorption experiments for As(V) and As(III) onto the surfaces of aerobic Pseudomonas aeruginosa, which can be readily isolated from natural media, were conducted under nutrient-absent conditions. While a small amount of As(III) was adsorbed on the bacterial cell surfaces, As(V) was not effectively removed from the solution through adsorption. The result was likely due to the electrostatic repulsion between anionic compounds of aqueous As(V) and cell surfaces of f aeruginosa. However, the bacteria forming biofilm reduced a large amount of aqueous As(V) to As(III), which indicated that microorganisms in most oligotrophic, natural geologic settings can mediate the behavior of aqueous As. Biobarriers designed to remove the various heavy metals in contaminant plume may practically lead to the enhancement of toxicity and mobility of As.