• Title/Summary/Keyword: fluorescent in situ hybridization(FISH)

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In situ Analysis of Methanogenic Bacteria in the Anaerobic Mesophilic and Thermophilic Sludge Digestion (중온 및 고온 혐기성 소화에서 메탄생성균 군집 분석에 관한 연구)

  • Hwang, Sun-Jin;Jang, Hyun-Sup;Eom, Hyoung-Choon;Jang, Kwang-Un
    • Journal of Korean Society of Water and Wastewater
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    • v.18 no.4
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    • pp.515-521
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    • 2004
  • Anaerobic digestion has many advantages over the more conventional aerobic treatment processes such as low levels of excess sludge production, low space (area) requirements, and the production of valuable biogas. The purpose of this study was to evaluate the effect of organic loading rate of anaerobic digestion on thermophilic($55^{\circ}C$) and mesophilic($35^{\circ}C$) conditions. Fluorescent in situ hybridization (FISH) method was also used to study the microbial community in the reactors. The stabilizing time in mesophilic anaerobic reactors was shorter as approximately 20 days than 40 days in the thermophilic anaerobic reactors. The amount of methane production rate in anaerobic reactors was independent of the concentrations of supplied substrates and the amount of methanogens. When the microbial diversity in the mesophilic and thermophilic reactors, which had been treated with acetate-based artificial wastewater, were compared, it was found that methanogenesis was carried out by microbial consortia consisting of bacteria and archaea such as methanogens. To investigate the activity of bacterial and archaeal populations in all anaerobic reactors, the amount of acetate was measured. Archaea were predominant in all reactors. Interestingly, Methanothrix-like methanogens appeared in mesophilic anaerobic reactors with high feed substrate concentrations, whereas it was not observed in thermophilic anaerobic reactors.

Microbial Characterization of Excessive Growing Biofilm in Sewer Lines Using Molecular Technique

  • LEE YOUNG-OK;PARK JIN-HWA;PARK JAE-K.
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.938-945
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    • 2005
  • For elucidating excessive growth of biofilm that subsequently leads to the clogging problem in a small town's sewer lines of Wisconsin, the FISH method was employed. At the beginning of the simulated experiments, ${\beta}$-subclass proteobacteria prevailed in runs fed with industrial wastewater, while ${\gamma}$-subclass proteobacteria dominated in runs with domestic wastewater. However, the bacterial community structure changed significantly over six weeks; Cytophaga-Flavobacterium (CF)­group bacteria dominated in most runs fed with the small town's wastewater regardless of their source, while CF-group decreased strongly in run fed with domestic sewage from another city (Madison). It was also microscopically confirmed that most of those clogging materials was toilet tissue, which in turn may lead to vigorous growth of cellulose-degrading CF-group bacteria. This dominant presence of CF-group bacteria in the small town's sewer indicates that the main constituent of biofilm, toilet tissue (cellulose) in sewage, might have induced the unique pattern of their microbial community structure. Therefore, it suggests that molecular technique is useful for monitoring the clogging problems in sewer lines.

Bacterial Community of Free-living and Aggregated Bacteria at Thawing Period in Lake Baikal (해빙기 바이칼호에서 부유세균과 Aggregates에 부착한 세균의 군집구조)

  • 홍선희;김옥선;전선옥;유재준;안태석
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.192-197
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    • 2002
  • Fluorescent in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes was used to compare the community structures of free-living and aggregated bacteria at thawing period in Lake Baikal. Targeted groups were Eubacteria, $\alpha$-, $\beta$-, $\gamma$- proteobacteria groups, Cytophaga-Flavobacterium group and Planctomycetales. Total bacterial numbers of free-living bacteria were ranged from $0.2{\times}10^6\cells{\cdot}ml^-1$ to $3.2{\times}10^6\cells{\cdot}ml^-1$, which were decreasing with depth, while the aggregated bacterial numbers were dramatically increasing from $0.4{\times}10^4 to 3.3{\times}10^4 \cells{\cdot}ml^-1$ with depth. The ratios of EUB probe binding cells to DAPI counts were ranged from 52.3 to 74.1% in free-living bacteria, and from 39.6 to 66.7% in the aggregated bacteria, respectively. Community structures of the aggregated bacteria were very different from each free-living bacteria at every depth. At 25 m depth, where the chlorophyll a concentration was highest, both structures were quite different from those of surface layers, rendering the fact that the community structures might be affected by phytoplankton. The vertical profile of community structure of aggregated bacteria is particular. The proportion of $\beta$-proteobacteria group was increasing with depth and it was 51.8% at 100 m, but the dominant group was $\gamma$-pro-teobacteria group at 250 m. Taken together, the biodiversity and succession of aggregated bacteria are quite different from free-living bacteria.

Nitrite Accumulation Characteristics and Quantitative Analyses of Nitrifying and Denitrifying Bacteria in a Sequencing Batch Reactor (연속회분반응기의 아질산 축적 특성과 질산화 및 탈질 미생물의 정량적 분포 연구)

  • Kim, Dong-Jin;Kwon, Hyun-Jin;Yoon, Jung-Yee;Cha, Gi-Cheol
    • Journal of Korean Society on Water Environment
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    • v.24 no.3
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    • pp.383-390
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    • 2008
  • Recently, the interests on economical nitrogen removal from wastewater are growing. As a method of the novel nitrogen removal technology, nitrogen removal via nitrite pathway by selective inhibition of free ammonia and free nitrous acid on nitrite oxidizing bacteria have been intensively studied. The inhibition effects of free ammonia and free nitrous acid are low when domestic wastewater is used, however, because of its relatively lower nitrogen concentration than the wastewater from industry and landfill, etc. In this study, a sequencing batch reactor (SBR) is proposed for nitrogen removal to investigate the effect of the low nitrogen concentration on nitrite accumulation. Nitrification efficiency reached almost 100% during the aerobic cycle and the maximum specific nitrification rate ($V_{max,nit}$) reached $17.8mg\;NH_4{^+}-N/g\;MLVSS{\bullet}h$. During the anoxic cycle, average denitrification efficiency reached 87% and the maximum specific denitrification rate ($V_{max,den}$) reached $9.8mg\;NO_3{^-}-N/g\;MLVSS{\bullet}h$. From the analysis the main reason of nitrite accumulation in the SBR was free nitrous acid rather than free ammonia. Nitrite accumulation increased with the decrease of organic content in the wastewater and the mechanism is not well understood yet. From the result of fluorescent in situ hybridization, the distribution of nitrite oxidizing bacteria was in equilibrium with ammonium oxidizing bacteria when nitrite accumulation did not occur.

Chromosome Aberrations in Porcine Embryo Produced by Nuclear Transfer with Somatic Cell

  • Ah, Ko-Seung;Jin, Song-Sang;Tae, Do-Jeong;Chung, Kil-Saeng;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.73-73
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    • 2002
  • Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

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Monitoring of Soil Bacterial Community and Some Inoculated Bacteria After Prescribed Fire in Microcosm

  • Song Hong-Gyu;Kim Ok-Sun;Yoo Jae-Jun;Jeon Sun-Ok;Hong Sun-Hee;Lee Dong-Hun;Ahn Tae-Seok
    • Journal of Microbiology
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    • v.42 no.4
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    • pp.285-291
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    • 2004
  • The soil bacterial community and some inoculated bacteria were monitored to assess the microbial responses to prescribed fire in their microcosm. An acridine orange direct count of the bacteria in the unburned control soil were maintained at a relatively stable level $(2.0\~2.7\times10^9\;cells/g^{-1}{\cdot}soil)$ during the 180 day study period. The number of bacteria in the surface soil was decreased by fire, but was restored after 3 months. Inoculation of some bacteria increased the number of inoculated bacteria sev­eral times and these elevated levels lasted several months. The ratios of eubacteria detected by a flu­orescent in situ hybridization (FISH) method to direct bacterial count were in the range of $60\~80\%$ during the study period, with the exception of some lower values at the beginning, but there were no definite differences between the burned and unburned soils or the inoculated and uninoculated soils. In the unburned control soil, the ratios of $\alpha-,\beta-\;and\;\gamma-subgroups$ of the proteobacteria, Cytophaga-Fla­vobacterium and other eubacteria groups to that of the entire eubacteria were 13.7, 31.7, 17.1, 16.8 and $20.8\%,$ respectively, at time 0. The overall change on the patterns of the ratios of the 5 subgroups of eubacteria in the uninoculated burned and inoculated soils were similar to those of the unburned con­trol soil, with the exception of some minor variations during the initial period. The proportions of each group of eubacteria became similar in the different microcosms after 6 months, which may indicate the recovery of the original soil microbial community structure after fire or the inoculation of some bac­teria. The populations of Azotobacter vinelandii, Bacillus megaterium and Pseudomonas fluorescens, which had been inoculated to enhance the microbial activities, and monitored by FISH method, showed similar changes in the microcosms, and maintained high levels for several months.

The Characteristics of Microbial Community for Biological Activated Carbon in Water Treatment Plant (생물활성탄 공정에서 활성탄 재질에 따른 부착미생물 군집특성)

  • Son, Hee-Jong;Park, Hong-Ki;Lee, Soo-Ae;Jung, Eun-Young;Jung, Chul-Woo
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.12
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    • pp.1311-1320
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    • 2005
  • The purpose of this research is to survey characteristics of microbial community and the removal efficiency of organic materials for biological activated carbon in water treatment plant. Coal based activated carbon retained more attached bacterial biomass on the surface of the activated carbon than the other activated carbon with operating time and materials. The heterotrophic plate count(HPC), eubacteria(EUB) and 4,6-diamidino-2-phenylindole(DAPI) counts were ranged from $0.95{\times}10^7$ to $52.4{\times}10^7$ CFU/g, from $3.8{\times}10^8$ to $134.2{\times}10^8$ cells/g and from $7.0{\times}10^8$ to $250.2{\times}10^8$ cells/g, respectively. The biomass of EUB and DAPI appeared to be much more $10^2$ than HPC, which were increasing in bed volume of 20,000 at the stage of steady-state. The change of microbial community by analyzing fluorescent in situ hybridization(FISH) method with rRNA-targeted oligonucleotide probes, the dominant group was $\alpha$-proteobacteria($\alpha$ group) and high G+C content bacteria(HGC) the lowest distributing rate before reaching the bed volume of 20,000. After reaching the bed volume of 20,000, $\alpha$ group and other groups of bacteria became decreased, on the other hand, the proportion of both $\beta$-proteobacteria($\beta$ group) and $\gamma$-proteobacteri($\gamma$ group) were increasing. Coconut and wood based activated carbons had similar trend with coal based activated carbon, but the rate of $\alpha$ group on coal based activated carbon had gradually increased. Bacterial production with the operating period appeared highest in coal based activated carbon at the range of $1.2{\sim}3.4\;mg-C/m^3{\cdot}h$ while the coconut and wood based activated carbon were ranged from 1.1 to 2.6 $mg-C/m^3{\cdot}h$ and from 0.7 to 3.5 $mg-C/m^3{\cdot}h$ respectively. The removal efficiency of assimilable organic carbon(AOC) showed to be highly correlated with bacterial production. The correlation coefficient between removal efficiency of AOC and bacterial production were 0.679 at wood based activated carbon, 0.291 at coconut based activated carbon and 0.762 at coal based activated carbon, respectively.

Analysis of Attached Bacterial Community of Biological Activated Carbon Process Using FISH (FISH 기법을 이용한 생물활성탄 공정에서의 운전기간별 부착 박테리아 군집변화 분석)

  • Son, Hyeng-Sik;Son, Hee-Jong;Park, Geun-Tae;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.22 no.1
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    • pp.25-35
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    • 2013
  • The concentration of organic compounds was analyzed at each step of BAC process though $BDOC_{total/rapid/slow}$. Further, bacteria communities and biomass concentrations measured FISH and ATP methods were analyzed. The bed volume (BV) of steady state is different from that of based on assessment of organic compounds removal. Bed volumes in DOC, $BDOC_{rapid}$ and $BDOC_{total/slow}$ removal at steady state were around 27,500 (185.8 day), 15,000 (101.4 day) and 32,000 (216.2 day), respectively. A biomass didn't change after the bed volume reached 22,500 (152.0 day) according to analyzing ATP concentration of bacteria. The concentration of ATP was 2.14 ${\mu}g/g$ in BV 22,500 (152.0 day). The total bacterial number was $4.01{\pm}0.4{\times}10^7$ cells/g at the bed volume 1,150 (7.8 day) (the initial operation) and the number of bacteria was $9.27{\pm}0.2{\times}10^9$ at the bed volume 58,560 395.7 day) that increased more than 200 times. Bacterial uptrend was reduced and bacterial communities were stabilized since BV 18,720 (126.5 day). When BV were 1,150 (7.8 day), 8,916 (60.2 day), 18,720 (126.5 day), 31,005 (209.5 day), 49,632 (335.3 day), 58,560 (395.7 day), a proportion of total bacteria for the Eubacteria were 60.1%, 66.0%, 78.4%, 82.0%, 81.3% respectively. ${\gamma}$-Proteobacteria group was the most population throughout the entire range. The correlation coefficient ($r^2$) between Eubacteria biomass and ATP concentration was 0.9448.

Abundance and Structure of Microbial Loop Components (Bacteria and Protists) in Lakes of Different Trophic Status

  • Chrost, Ryszard J.;Tomasz, Adamczewski;Kalinowska, Krystyna;Skowronska, Agnieszka
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.858-868
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    • 2009
  • The abundance, biomass, size distribution, and taxonomic composition of bacterial and protistan (heterotrophic and autotrophic nanoflagellates and ciliates) communities were investigated in six lakes of Masurian Lake District (north-eastern Poland) differing in trophic state. Samples were taken from the trophogenic water layer during summer stratification periods. Image analysis techniques with fluorescent in situ hybridization (FISH) as well as [$^3H$]-methyl-thymidine incorporation methods were applied to analyze differences in the composition and activity of bacterial communities. The greatest differences in trophic parameters were found between the humic lake and remaining non-humic ones. The same bacterial and heterotrophic nanoflagellate (HNF) cell size classes dominated in all the studied lakes. However, distinct increases in the contributions of large bacterial (>$1.0{\mu}m$) and HNF (>$10{\mu}m$) cells were observed in eutrophic lakes. The bacterial community was dominated by the ${\beta}$-Proteohacteria group, which accounted for 27% of total DAPI counts. Ciliate communities were largely composed of Oligotrichida. Positive correlations between bacteria and protists, as well as between nanoflagellates (both heterotrophic and autotrophic) and ciliates, suggest that concentrations of food sources may be important in determining the abundance of protists in the studied lakes.

A chromosome 1q44 deletion in a 4-month-old girl; The first report in Korea

  • Cho, Joo Hyun;Song, Eun Song;Kim, Hee Na;Oh, Burm Seok;Choi, Young Youn
    • Clinical and Experimental Pediatrics
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    • v.57 no.6
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    • pp.292-296
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    • 2014
  • The deletion of the distal long arm of chromosome 1 is associated with a characteristic facial appearance and a pattern of associated malformations. Characteristic manifestations include a round face with prominent 'cupid's bow' and downturned corners of the mouth, thin vermilion borders of lips, a long upper lip with a smooth philtrum, a short and broad nose, epicanthal folds, apparently low-set ears, micrognathia, microcephaly, abnormal hands and feet, variable cardiac or genital anomalies, moderate to severe mental retardation, and growth retardation. Using fluorescent in situ hybridization (FISH) analysis to map precisely the deletion, we present a case of chromosome 1q44 deletion with craniofacial characteristics, multiple congenital anomalies, and growth and psychomotor retardation. In comparison with other reported cases of 1q43-44 deletion, the subject does not show hydrocephalus, seizure, syn- or polydactyly of hands, and a urogenital anomaly. However, an arachnoid cyst, pinpoint dimple on the midline of the forehead, a right-sided supernumerary nipple and auricular pit, polydactyly of the right foot, adducted thumb, and flexion restriction of the proximal interphalangeal joint with a simian line in both hands were observed additionally.