• Title/Summary/Keyword: sediment incubation

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Polychlorobiphenyl (PCB) 토양오염복원: PCB 제거 토양미생물들의 군집과 기능을 효과적으로 분석하는 신 genomics 방법개발에 관한 연구

  • Park Jun-Hong
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2005.04a
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    • pp.28-30
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    • 2005
  • Because of high population diversity in soil microbial communities, it is difficult to accurately assess the capability of biodegradation of toxicant by microbes in soil and sediment. Identifying biodegradative microorganisms is an important step in designing and analyzing soil bioremediation. To remove non-important noise information, it is necessary to selectively enrich genomes of biodegradative microorganisms fromnon-biodegradative populations. For this purpose, a stable isotope probing (SIP) technique was applied in selectively harvesting the genomes of biphenyl-utilizing bacteria from soil microbial communities. Since many biphenyl-using microorganisms are responsible for aerobic PCB degradation In soil and sediments, biphenyl-utilizing bacteria were chosen as the target organisms. In soil microcosms, 13C-biphenyl was added as a selective carbon source for biphenyl users, According to $13C-CO_2$ analysis by GC-MS, 13C-biphenyl mineralization was detected after a 7-day of incubation. The heavy portion of DNA(13C-DNA) was separated from the light portion of DNA (12C-DNA) using equilibrium density gradient ultracentrifuge. Bacterial community structure in the 13C-DNAsample was analyzed by t-RFLP (terminal restriction fragment length polymorphism) method. The t-RFLP result demonstates that the use of SIP efficiently and selectively enriched the genomes of biphenyl degrading bacteria from non-degradative microbes. Furthermore, the bacterial diversity of biphenyl degrading populations was small enough for environmental genomes tools (metagenomics and DNA microarrays) to be used to detect functional (biphenyl degradation) genes from soil microbial communities, which may provide a significant progress in assessing microbial capability of PCB bioremediation in soil and groundwater.

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Analysis of Microbial Community During the Anaerobic Dechlorination of Perchloroethylene and Trichloroethylene (Perchloroethylene과 Trichloroethylene의 혐기적 탈염소화 및 미생물 군집 분석)

  • Lee Jae-Won;Kim Byung-Hyuk;Ahn Chi-Yong;Kim Hee-Sik;Yoon Byung-Dae;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.281-286
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    • 2005
  • In this study, the anaerobic enrichment cultivation was performed with the sediments and the dredged soils from the cities of Ulsan, Masan, Yeosu, Gwangyang, Ansan and Seongnam. Acetate as an electron donor and PCE (perchloroethylene) or TCE (trichloroethylene) as an electron acceptor were injected into the serum bottle with an anaerobic medium. After the incubation of 12 weeks, the removal efficiency of PCE was highest at $70\%$ in the treatment with the sediment of Ulsan. Also, the bacterial community structure was analyzed by D-DGGE (double denatured gradient gel electrophoresis) through PCR-based 16S rDNA approaches. The dominant species id the anaerobic enrichment were found to belong to the genus of Desulfovibrio.

Analysis of Microbial Community During the Anaerobic Dechlorination of Tetrachloroethylene (PCE) in Stream of Gimpo and Inchon Areas (경기도 김포, 인천 서구지역 소하천의 PCE 탈염소화 군집의 선별 및 다양성 분석)

  • Kim, Byung-Hyuk;Baek, Kyung-Hwa;Cho, Dea-Hyun;Sung, Youl-Boong;Ahn, Chi-Yong;Oh, Hee-Mock;Koh, Sung-Cheol;Kim, Hee-Sik
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.140-147
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    • 2009
  • In this study, anaerobic enrichment cultivation was performed with the sediments from the Gimpo and Inchon areas. Lactate as an electron donor and PCE as an electron acceptor was injected into the serum bottle with an anaerobic medium. After the incubation of 8 weeks, the reductive dechlorination of PCE was observed in 7 sites among 16 sites (43%). Three enrichment cultures showed completely dechlorination of PCE to ethene, while four enrichment culture showed transformation of PCE to cis-DCE. The bacterial community structure was analyzed by PCR-DGGE. Dechlorinating bacteria were detected by species-specific primers. The dominant species in seven anaerobic enrichments were found to belong to the genus of Dehalococcoides sp. and Geobacter sp., and Dehalobacter sp.

Evaluation of Petroleum Oil Degrading Mixed Microorganism Agent for the Bioremediation of Petroleum Oil Spilled in Marine Environments (해양유류오염정화를 위한 유류분해 미생물제제의 평가)

  • Sohn, Jae-Hak
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1599-1606
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    • 2011
  • To evaluate the effects of microorganism agents on oil biodegradation, treatability and microcosm studies were conducted. Petroleum oil degrading bacteria were isolated from enriched cultures of oil-contaminated sediment samples using a mineral salts medium (MSM) containing 0.5% Arabian heavy crude oil as the sole carbon source. After a 5 day-incubation period using MSM, mixed microorganisms of three species (strains BS1, BS2 and BS4) degraded 48.4% of aliphatic hydrocarbons and 30.5% of aromatic hydrocarbons. Treatability and microcosm tests were performed in the three different treatment conditions (AO: Arabian heavy crude oil, AO+IN: Arabian heavy crude oil+inorganic nutrient, AO+IN+MM: Arabian heavy crude oil+inorganic nutrient+mixed microorganism agents). Among these, significantly enhanced biodegradation of aliphatic hydrocarbons were observed in AO+IN and AO+IN+MM conditions, without showing any different biodegradation rates in either condition. However, the degradation rates of aromatic hydrocarbons in an AO+IN+MM condition were increased by 50% in the treatability test and by 13% in the microcosm test compared to those in an AO+IN condition. Taken together, it can be concluded that mixed microorganism agents enhance the biodegradation of aliphatic and aromatic hydrocarbons in laboratory, a treatability test, and a microcosm test. This agent could especially be a useful tool in the application of bioremediation for removal of aromatic hydrocarbons.

Electron-microscopic studies on fine structure and enzyme activity in the axenic and conventional strains of Entamoeba histolytica (이질아메바(Entamoeba histolytica)의 미세구조 및 효소활성에 관한 전자현미경적 연구)

  • Yong, Tae-Sun;Jeong, Pyeong-Rim;Lee, Geun-Tae
    • Parasites, Hosts and Diseases
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    • v.23 no.2
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    • pp.269-284
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    • 1985
  • The metabolism of Entamoeba histolytica would be affected by various environmental factors, and alteration of the environment was known to afEect the fine structure of 5. histolytica. The present study was designed electronmicroscopically to investigate the ultrastructure and enzyme activities in the aEonic and conventional strains of 5. histolytica. The trophozoites of axenically cultivated HK-9 strain and conventional YS-27 and YS-49 strains of 5. histolytica were collected and liKed with 4% paraformaldehyde/0.1M cacodylate buffier(pH 74), After washing them by centrifugation, 1% warm agar was added in the sediment. Solidified agar with the trophozoites was cut into $lmm^3$ cubes, and incubated in the various substrates to observe enzyme activities. Then, the specimen was post-fixed with 3% glutaraldehyde/0.1M cacodylate buffer (PH 7.4) and 1% osmium tetroBide/0.1M cacodylate buffier (pH 7.4) , dehydrated in ascending ethanol series and embedded in epoxy resin. These were sectioned on an ultramicrotome and observed with a transmission electronmicroscope. The procedures for the observation of the fine structure were same as the above, except for the incubation in the substrate. The sections were stained with uranyl acetate and lead citrate. For the observation of the surface of the amoebae, scanning-electronmicroscopy was carried out. The results obtained in the present study are summarized as follows: 1. The fuzzy coat around double-layered plasma membrane of 5. histolytica was more irregularly and densely distributed in the conventional strains (YS-27, YS-49 strains) than in the axonic strain (HK-9 strain). 2. The endosomes, button bodies and chromatin material were surrounded by a double-layered nuclear membrane having scattered nuclear fores. The paranuclear body, mono- or double-layered vacuoles, vacuolar membrane whorls, rosette-like cylindrical bodies, aggregation of cylindrical bodies and helical bodies were found in the cytoplasm of the amoebae. Helical bodies and glycogen granules were generally abundant, while a few smooth endoplasmic reticula were observed in the cytoplasm. 3. Alkaline phosphatase activity was mainly demonstrated in the plasma membrane, limiting membranes of vacuoles and smooth endoplasmic reticula. ATPase activity was observed in the nucleus, limiting membranes of vacuoles and vacuolar membrane whorls. 4. Acid phosphatase activity was commonly demonstrated in the limiting membranes an contents of vacuoles, Iysosome-like organelles, plasma membrane and the button bodies in the nucleus. The activity was more weakly demonstrated in the HK-9 strain than in the other conventional strains of 5. histolytica. No peroBidase activity was observed in the amoeba strains employed in the present study. 5. With a scanning electron-microscope, no distinct structural differences were observed between the amoeba strains. All the trophozoite forms of the amoebae showed crater-like depressions and rugged features on the outer surface.

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Analysis of Microbial Community during the Anaerobic Dechlorination of PCE/TCE by DGGE (DGGE를 이용한 PCE 및 TCE의 혐기적 탈염소화 군집의 미생물 군집분석)

  • Kim, Byung-Hyuk;Cho, Dae-Hyun;Sung, Youl-Boong;Ahn, Chi-Youg;Yoon, Byung-Dae;Koh, Sung-Cheol;Oh, Hee-Mock;Kim, Hee-Sik
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.448-454
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    • 2010
  • This study investigated the effect of PCE and TCE as electron acceptors on the bacterial composition of dechlorinating communities. The enrichment cultures reductively dechlorinating PCE and TCE were developed from three environment samples using acetate as electron donor. The cultures were prepared by sequential enrichment, which was seeded with sediment and dredged soil. Denatured gradient gel electrophresis (DGGE) of 16S rRNA gene fragment was used to compare the microbial communities of these three enrichment cultures. After incubation for 4 weeks, the removal efficiencies of PCE and TCE were highest from Yeocheon site (87.37% and 84.46%, respectively). PCE and TCE as electron acceptors affected the bacterial diversity and community profiles in the enrichment cultures. DGGE analysis showed that the dominant bacteria in PCE and TCE enrichment were belonged to Clostridium sp., Desulfotomaculum sp., and uncultured bacteria.

Analysis of Nitrogen and Phosphorus Benthic Diffusive Fluxes from Sediments with Different Levels of Salinity (염분농도에 따른 호소 퇴적물 내 질소 및 인 용출 특성 분석)

  • Seulgi Lee;Jin Chul Joo;Hee Sun Moon;Dong Hwi Lee;Dong Jun Kim;Jiwon Choi
    • Ecology and Resilient Infrastructure
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    • v.10 no.3
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    • pp.85-96
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    • 2023
  • The study involved the categorization of domestic lakes located in South Korea into three groups based on their salinity levels: upstream reservoirs with salinity less than 0.3 psu, estuarine reservoirs with salinity ranging from 0.3 to 2 psu, and brackish lagoons with salinity exceeding 2 psu. Subsequently, the research assessed variations in the concentrations of total nitrogen (T-N) and total phosphorus (T-P) in the sediment of these lakes using statistical analysis, specifically one-way analysis of variance (ANOVA). Additionally, a laboratory core incubation test was conducted to investigate the benthic nutrient fluxes in Songji lagoon (salinity: 11.80 psu), Ganwol reservoir (salinity: 0.73 psu), and Janggun reservoir (salinity: 0.08 psu) under both aerobic and anoxic conditions. The findings revealed statistically significant differences in the concentrations of T-N and T-P among sediments in the lakes with varying salinity levels (p<0.05). Further post-hoc analysis confirmed significant distinctions in T-N between upstream reservoirs and estuarine reservoirs (p<0.001), as well as between upstream reservoirs and brackish lagoons (p<0.01). For T-P, a significant difference was observed between upstream reservoirs and brackish lagoons (p<0.01). Regarding benthic nutrient fluxes, Ganwol Lake exhibited the highest diffusive flux of NH4+-N, primarily due to its physical characteristics and the inhibition of nitrification resulting from its relatively high salinity. The flux of NO3--N was lower at higher salinity levels under aerobic conditions but increased under anoxic conditions, attributed to the impact of salinity on nitrification and denitrification. Additionally, the flux of PO43--P was highest in Songji Lake, followed by Ganwol Lake and Janggun Reservoir, indicating that salinity promotes the diffusive flux of phosphate through anion adsorption competition. It's important to consider the influence of salinity on microbial communities, growth rates, oxidation-reduction processes, and nutrient binding forms when studying benthic diffusive nutrient fluxes from lake sediments.

The Development of a Benthic Chamber (BelcI) for Benthic Boundary Layer Studies (저층 경계면 연구용 Benthic chamber(BelcI) 개발)

  • Lee, Jae-Seong;Bahk, Kyung-Soo;Khang, Buem-Joo;Kim, Young-Tae;Bae, Jae-Hyun;Kim, Seong-Soo;Park, Jung-Jun;Choi, Ok-In
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.41-50
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    • 2010
  • We have developed an in-situ benthic chamber (BelcI) for use in coastal studies that can be deployed from a small boat. It is expected that BelcI will be useful in studying the benthic boundary layer because of its flexibility. BelcI is divided into three main areas: 1) frame and body chamber, 2) water sampler, and 3) stirring devices, electric controller, and data acquisition technology. To maximize in-situ use, the frame is constructed from two layers that consist of square cells. All electronic parts (motor controller, pA meter, data acquisition, etc.) are low-power consumers so that the external power supply can be safely removed from the system. The hydrodynamics of BelcI, measured by PIV (particle image velocimetry), show a typical "radial-flow impeller" pattern. Mixing time of water in the chamber is about 30 s, and shear velocity ($u^*$) near the bottom layer was calculated at $0.32\;cm\;s^{-1}$. Measurements of diffusivity boundary layer thickness showed a range of $180-230\;{\mu}m$. Sediment oxygen consumption rate, measured in-situ,was $84\;mmol\;O_2\;m^{-2}\;d_{-1}$, more than two times higher than on-board incubation results. Benthic fluxes assessed from in-situ incubation were estimated as follows: nitrate + nitrite = $0.18\;{\pm}\;0.07\;mmol\;m^{-2}\;d^{-1}$ ammonium $23\;{\pm}\;1\;mmol\;m^{-2}\;d^{-1}$ phosphate = $0.09\;{\pm}\;0.02\;mmol\;m^{-2}\;d^{-1}$ and silicate = $23\;{\pm}\;1\;mmol\;m^{-2}\;d^{-1}$.

Studies on canine babesiosis in Korea I. In vitro isolation and antigenic properties of Babesia gibsoni (개 바베시아병에 관한 연구 I. Babesia gibsoni의 시험관내 분리와 항원성상에 관한 연구)

  • Lee, Ho-kweon;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.681-692
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    • 1996
  • The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

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Combined Effects of Filter-feeding Bivalve and Zooplankton on the Growth Inhibition of Cyanobacterium Microcystis aeruginosa (남세균 제어를 위한 동물플랑크톤(Daphnia magna)과 패류(Unio douglasiae)의 단독 및 혼합적용)

  • Kim, Nan-Young;Park, Myung-Hwan;Hwang, Su-Ok;Kim, Baik-Ho;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.48 no.2
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    • pp.108-114
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    • 2015
  • Single - and combined effects of a domestic freshwater bivalve Unio douglasiae (7.6~8.6 cm in shell length) and zooplankton Daphnia magna (1~2 mm in body size) were examined to understand whether they inhibit the growth of harmful cyanobacterial bloom (i.e. Microcystis aeruginosa) in a eutrophic lake. The experiments were triplicated with twelve glass aquaria (40 L in volume); three aquaria without mussel and zooplankton, served as a control, three zooplankton aquaria (Z, density=40 indiv. $L^{-1}$), three mussel aquaria (M, density=0.5 indiv. $L^{-1}$), and three mussel plus zooplankton aquarium (ZM, density=40 indiv.Z $L^{-1}$ plus 0.5 indiv.M/L), respectively. Algal growth inhibition (%) calculated as a difference in the concentration of chlorophyll-a (Chl-a) before and after treatment. Chl-a in all aquaria decreased with the time, while a greatest algal inhibition was seen in the ZM aquaria. After 24 hrs of incubation, Chl-a concentration at the mid-depth (ca. 15 cm) in ZM aquaria reduced by 90.8% of the control, while 63.2% and 79.8% in Z and M aquaria, respectively. Interestingly, during the same period, the surface Chl-a was diminished by 51.9% and 65.4% relative to the control in Z and ZM aquaria, while 27.4% of initial concentration decreased in M aquarium, respectively. These results suggest that 1) this domestic freshwater filter-feeding bivalve plays a significant role in the control of cyanobacterial bloom (M. aeruginosa), and 2) the combination with zooplankton and mussel has a synergistic effect to diminish them, compared to the single treatment of zooplankton and mussel.