• 제목/요약/키워드: Denaturing gradient gel electrophoresis (DGGE)

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한국산 꽃소금과 천일염의 이화학적 특성 및 미생물 분석 (Physicochemical Properties and Microbial Analysis of Korean Solar Salt and Flower of Salt)

  • 이혜미;이우경;진중현;김인철
    • 한국식품영양과학회지
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    • 제42권7호
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    • pp.1115-1124
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    • 2013
  • 한국산 꽃소금의 특성을 알아보기 위하여 국내에서 생산된 꽃소금 3종과 천일염 1종의 이화학적 및 미생물 분석을 실시하였다. 수분함량은 $10.54{\pm}0.10{\sim}13.82{\pm}0.12%$, 나머지 조단백, 조지방, 조섬유는 거의 존재하지 않았다. NaCl은 신의도산 꽃소금이 $78.81{\pm}0.28%$, 비금산 꽃소금이 $81.67{\pm}0.34%$, 도초산 꽃소금 $84.61{\pm}0.21%$, 도초산 천일염이 $80.82{\pm}0.17%$로 나타났다. 불용분, 사분은 각각 0.01~0.05%, 0.01~0.03%로 낮게 검출되었다. 미네랄 분석은 도초산 꽃소금에서 K과 Mg 함량이 2,975.23 mg/kg, 9,886.72 mg/kg으로 비교적 낮게 나타났다. Ca은 도초산 소금 2종이 945.53 mg/kg, 942.43 mg/kg으로 낮은 함량을 보였다. 중금속 As, Cd, Pb, Hg은 모두 규격 이하로 검출되었다. 소금결정을 관찰한 결과로 천일염과 꽃소금 모두 핵이 중복되어 겹으로 적층된 것을 확인하였으며 크기는 비금산 꽃소금이 $0.067{\times}0.067mm$로 가장 작고 도초산 천일염이 $0.112{\times}0.124mm$로 꽃소금에 비해 크게 나타났다. 소금의 색도는 꽃소금의 L값이 천일염에 비해 높아 밝게 확인되었다. 관능검사 결과로 짠맛은 NaCl 함량과 유사하며, 쓴맛은 K과 Mg 함량이 적은 도초산 꽃소금이 낮게 나타났다. 천일염에 비해 꽃소금의 단맛과 기호도가 더 높은 것으로 확인되었다. 호염균 동정 결과 19종 모두 Firmicutes로 꽃소금에서 Marinibacillus, Paenibacillus, Bacillus 속이 12종으로 확인되며, 천일염은 Planococcus, Staphylococcus, Bacillus 속 3종으로 검출되었다. DGGE 실험 결과로 도초산 꽃소금에서 16개의 band와 도초산 천일염에서 15개의 band를 확인하였다. 동정 결과 도초산 꽃소금에서 Cupriavidus sp. ATHA3(14.43%), Cupriavidus sp. TSA5(10.41%), Maritimibacter sp. YCSD61-2(8.13%), uncultured bacteria(68%)로 확인되었고, 도초산 천일염에서는 Dunaliella salina(4.76%), Cupriavidus sp. ATHA3(15.80%), uncultured Mycoplasmataceae bacteria(51.72%), uncultured bacteria(27%)로 확인되었다.

Stability of Partial Nitrification and Microbial Population Dynamics in a Bioaugmented Membrane Bioreactor

  • Zhang, Yunxia;Xu, Yanli;Jia, Ming;Zhou, Jiti;Yuan, Shouzhi;Zhang, Jinsong;Zhang, Zhen-Peng
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1656-1664
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    • 2009
  • Bioaugmentation of bioreactors focuses on the removal of numerous organics, with little attention typically paid to the maintenance of high and stable nitrite accumulation in partial nitrification. In this study, a bioaugmented membrane bioreactor (MBR) inoculated with enriched ammonia-oxidizing bacteria (AOB) was developed, and the effects of dissolved oxygen (DO) and temperature on the stability of partial nitrification and microbial community structure, in particular on the nitrifying community, were evaluated. The results showed that DO and temperature played the most important roles in the stability of partial nitrification in the bioaugmented MBR. The optimal operation conditions were found at 2-3 mgDO/l and $30^{\circ}C$, achieving 95% ammonia oxidization efficiency and nitrite ratio ($NO_2^-/{NO_x}^-$) of 0.95. High DO (5-6 mg/l) and low temperature ($20^{\circ}C$) had negative impacts on nitrite accumulation, leading to nitrite ratio drop to 0.6. However, the nitrite ratio achieved in the bioaugmented MBR was higher than that in most previous literatures. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) were used to provide an insight into the microbial community. It showed that Nitrosomonas-like species as the only detected AOB remained predominant in the bioaugmented MBR all the time, and coexisted with numerous heterotrophic bacteria. The heterotrophic bacteria responsible for mineralizing soluble microbial products (SMP) produced by nitrifiers belonged to the Cytophaga-Flavobacterium-Bacteroides (CFB) group, and $\alpha$-, $\beta$-, and $\gamma$- Proteobacteria. The fraction of AOB ranging from 77% to 54% was much higher than that of nitrite-oxidizing bacteria (0.4-0.9%), which might be the primary cause for the high and stable nitrite accumulation in the bioaugmented MBR.

Impact of phosphorus application on the indigenous arbuscular mycorrhizal fungi, soybean growth and yield in a 5-year phosphorus-unfertilized crop rotation

  • Higo, Masao;Sato, Ryohei;Serizawa, Ayu;Gunji, Kento;Suzuki, Daisuke;Isobe, Katsunori
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.351-351
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    • 2017
  • Arbuscular mycorrhizal fungi (AMF) are particular soil fungi that benefit many crops and require a symbiosis with plant roots to survive. In our previous study, there was a positive correlation between AMF root colonization and soybean grain yield in a four-year consecutive winter cover crop-soybean rotational system without phosphorus fertilizer. It is suggested that higher AMF root colonization can be a better solution for improving soybean growth and grain yield in P-limited soil. Our purpose in this study was to test the hypothesis that a P application is the main factor improving soybean growth, P nutrition and grain yield, and the benefit from AMF to soybean P uptake and growth in a P-limited soil. Impact of a P application on AMF root colonization and communities in soybean roots and their potential contribution to soybean growth and P nutrition under a five-year P-unfertilized crop rotational system were investigated over two-years. In this study, four cover crop treatments included 1) wheat (Triticum aestivum); 2) red clover (Trifolium pratense); 3) rapeseed (Brassica napus); and 4) fallow in the crop rotation. The amount of triple superphosphate as a P fertilizer applied rate after cultivation of cover crops was 120 and $360k\;ha^{-1}$ in 2014 and 2015, respectively. Soybean roots were sampled at full-flowering and analyzed for AMF communities using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real-time PCR (qPCR) techniques. The AMF root colonization in the soybean roots at full bloom stage was significantly influenced by cover crop and P application throughout the two-year rotation. The two-year rotation of different cover crops or fallow impacted the molecular diversity of AMF communities colonizing roots of soybean. Redundancy analysis (RDA) indicated that AMF communities colonizing roots of soybean were significantly different among cover crop rotations. The AMF communities colonizing roots of soybean were clearly influenced by a P application in the two-year trial. Moreover, a P application may have positively impacts on the AMF communities under P-deficit soil due to the continuous cover crop-soybean rotational system without a P fertilizer.

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Microbial Floral Dynamics of Chinese Traditional Soybean Paste (Doujiang) and Commercial Soybean Paste

  • Gao, Xiuzhi;Liu, Hui;Yi, Xinxin;Liu, Yiqian;Wang, Xiaodong;Xu, Wensheng;Tong, Qigen;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • 제23권12호
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    • pp.1717-1725
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    • 2013
  • Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

상향류 활성탄 생물막 공정을 이용한 정체 수역 수질 개선 및 공정 내 미생물 군집 해석 (Water Quality Improvement of Stagnant Water using an Upflow Activated Carbon Biofilm Process and Microbial Community Analysis)

  • 오유미;이재호;박정진;최기충;박태주;이태호
    • 대한환경공학회지
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    • 제32권1호
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    • pp.23-32
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    • 2010
  • 정체수역에서는 자연적 흐름의 차단으로 인해 자정능력이 떨어지며, 영양염류의 축적으로 인해 부영양화와 같은 문제점이 발생한다. 또한 비점오염물질의 유입은 정체수역 내 난분해성 물질을 증가시킨다. 본 연구에서는 정체수역의 수질개선을 위해 무산소조, 호기1조, 호기2조로 구성된 장치형 상향류 활성탄 생물막 반응기를 도입하여 정체수의 연속적 순환에 따른 오염물질 농도의 변화를 모니터링 하였다. 정체수역을 모사하기 위하여 $2m^3$의 저장탱크에 유원지의 호소수를 저장하였으며, 수질개선을 위한 최적 유입 유량을 산출하기 위하여 HRT가 6 hr, 4 hr, 2 hr 가 되도록 호소수의 유입 유량을 변화시켰다. 이 가운데 HRT 4 hr에서 SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$, TN, TP의 제거 효율이 각각 69.8, 83.0, 91.3, 74.1, 74.7, 88.9%로 가장 좋은 수질 개선 효과를 얻을 수 있었다. 이에 HRT를 4 hr로 고정하고 골프장 연못수를 운전했을 때 SS, $BOD_5$, $COD_{Mn}$, $COD_{Cr}$ TN, TP의 제거 효율이 각각 78.5, 78.0, 80.2, 74.9, 55.6, 97.5% 달성되었다. 각 조건에서의 미생물 군집 변화를 PCR-DGGE를 사용하여 분석 결과, 유입수를 골프장 연못수로 교체함에 따라 미생물 군집에 변화가 나타났다. 또한 FISH에 의해 유입 유량 변화에 따른 질산화 미생물량의 변화를 관찰한 결과, HRT 4 hr의 조건에서 질산화 미생물이 가장 우점화됨을 알 수 있었다. 미생물량 및 INT-DHA를 이용한 미생물 활성도 실험 결과, HRT를 낮게 유지하였을 때에도 감소되지 않았다. 따라서 상향류 활성탄 생물막 공정을 정체 수역의 효과적인 수질 개선에 충분히 적용할 수 있을 것으로 기대한다.

Bacterial Community Dynamics during Swine In vitro Fermentation Using Starch as a Substrate with Different Feed Additives for Odor Reduction

  • Alam, Md.J.;Jeong, C.D.;Mamuad, L.L.;Sung, H.G.;Kim, D.W.;Cho, S.B.;Lee, K.;Jeon, C.O.;Lee, Sang-S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권5호
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    • pp.690-700
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    • 2012
  • The experiment was conducted by in vitro fermentation and bacterial community analysis to investigate the reduction of odorous compounds in response to the use of feed additives (FA) during carbohydrate overload in growing pigs. Soluble starch at 1% (control) and various FA at 0.1% Ginseng meal (FA1); Persimmon leaf (FA2); Gingko nut (FA3) and Oregano lippia (FA4) were added to fecal slurry and incubated anaerobically for 12 and 24 h. In vitro parameters and microbial diversity of the dominant bacteria following fermentation were analyzed using Denaturing Gradient Gel Electrophoresis (DGGE), band cloning and sequencing of the V3 region. Results showed that total gas production increased with the advancement of incubation (p<0.05). pH values of FAs and control groups were decreased except the FA4 group which increased somewhat from 12 to 24 h (p<0.05). Ammonia nitrogen ($NH_3$-N) and $H_2S$ gas concentrations were comparatively lower in both stages in FA4 treatment than in the other groups (p<0.05). Hence, $NH_3$-N concentrations in liquid phases were increased (p<0.05) from 12 to 24 h, but the trend was lowest in FA4 than in the other groups at both stages. The total VFA production was comparatively lower and butyrate levels were moderate in FA4 group than in the the other groups during both stages (p<0.05). Indirect odor-reducing compounds such as $NO_2$, $NO_3$ and $SO_4$ concentrations were higher in the FA4 and FA3 than in the other groups at 24 h (p<0.05). After fermentation, ten dominant bands appeared, six of which appeared in all samples and four in only the FA4 treated group. The total number of DGGE bands and diversity was higher in the FA4-group compared to other groups. Additionally, similarity indices were lowest (71%) in the FA4, which represented a different bacterial community compared with the other groups. These findings indicate that $NH_3$-N, $H_2S$ and VFA production was minimal, and pH was also better in the FA4 group than in the other groups. Furthermore, the conversion of odor-reducing indirect compounds or their intermediates was higher in the FA4 group in compared to the other groups. FA4 group generated less odorous products and more indirect products by in vitro fermentation at 24 h, and their microbial pattern appeared to differ from that of the other groups. These findings suggest that this particular FA could change the microbial population, which may have a beneficial effect on odor reduction. It is recommended that the oregano lippia may be supplied to growing pigs as FA along with excess carbohydrate sources to reduce the production of odorous compounds.

펜톤산화 및 생물학적 연속처리를 통한 유류오염토양의 효율적 처리 (Efficient Remediation of Petroleum Hydrocarbon-Contaminated Soils through Sequential Fenton Oxidation and Biological Treatment Processes)

  • 배재상;김종향;최정혜;칼루 이베 엑페게어;김수곤;고성철
    • 미생물학회지
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    • 제47권4호
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    • pp.356-363
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    • 2011
  • 유류의 유출로 인한 총석유계탄화수소(total petroleum hydrocarbons: TPH)는 종종 토양과 지하수의 오염을 초래하고 있다. TPH는 환경에 노출이 될 경우 물리화학적 과정을 거쳐 분해가 되나 그 반응은 상대적으로 느리다. 본 연구에서는 TPH로 오염된 토양의 환경친화적인 처리기법을 궁극적으로 개발하기 위해서 화학적 및 생물학적 통합기술을 도입하고자 시도하였다. 여기서 펜톤유사반응을 전처리단계로 도입하고 이후 디젤분해 혼합균을 처리하여(생물증강법) 오염유류를 처리하고자 하였다. 계면활성제 OP-10S (0.05%)과 산화제($FeSO_4$ 4%, 및 $H_2O_2$ 5%)를 사용할 경우 토양으로부터 효율적으로 TPH를 처리, 제거할 수 있는 것으로 나타났다. 디젤분해 혼합균을 토양슬러리에 접종할 경우 100배 이상 분해균의 밀도상승이 관찰되었는데 이는 접종된 분해균이 오염된 토양에서 성공적으로 존재할 수 있음을 의미한다($10^8-10^9$ CFU/g slurry). Fenton으로 처리된 토양에서의 TPH 제거 효율은 분해균으로 생물증강을 실시할 경우 최소한 57% 정도 상승되는 것으로 나타났다. 그러나 화학적, 생물학적 연속처리를 실시할 경우 대조구(무처리; 재거효율 95%)에 비해 상대적으로 낮은 처리효율(79-83%)을 나타내었는데, 이는 화학처리 중에 발생하는 자유기(free radicals) 함유 산화물질이 분해를 억제한 것에 기인하는 것으로 보인다. 본 연구에서의 얻어진 결과는 환경에 있어서 TPH로 오염된 토양과 저질을 효율적으로 정화하고 토양생태계의 신속한 회복에 활용할 수 있을 것으로 판단된다.