• 제목/요약/키워드: DGGE

검색결과 273건 처리시간 0.026초

Bacterial Diversity at Different Sites of the Digestive Tract of Weaned Piglets Fed Liquid Diets

  • Hong, Tran Thi Thu;Passoth, Volkmar;Lindberg, Jan Erik
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권6호
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    • pp.834-843
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    • 2011
  • Bacterial diversity was studied using PCR-DGGE, cloning and sequencing. DNA was isolated from digesta samples from stomach, ileum and colon of 28 weaned piglets (Large White${\times}$Mong Cai) fed dry control feed, naturally fermented liquid feed (FE) and a liquid diet with inclusion of rice distiller's residue feed. General bacterial diversity was described using DGGE analysis of the V3 region of 16S rDNA. The microbial populations in the stomach and the ileum were considerably influenced by the diet, while only marginal effects were observed in the colon. There was a large variation of the microbial flora in the stomach between individuals fed non-fermented diets. In contrast, animals fed diet FE had a more uniform microbial flora in the stomach and the ileum compared to the other diets. In total 47 bands from the DGGE profiles were cloned. In stomach, most frequently lactic acid bacteria were found. Feeding diet FE resulted in the occurrence of Pediococcus species in stomach and ileum. In pigs fed the other diets, Lactobacillus gallinarum, Lactobacillus johnsonii and Lactobacillus fermentum were found in stomach and ileum. Most of the sequences of bands isolated from colon samples and several from ileum matched to unknown bacteria, which often grouped within Prevotellaceae, Enterobacteriaceae, Bacteroidaceae and Erysipelotrichaceae. This study demonstrates that fermented liquid feed affects bacterial diversity and the specific microflora in stomach and ileum, which provides a potential to modulate the gut microflora with dietary means to increase the abundance of beneficial bacteria and improve piglets' health.

Microbial Diversity during Fermentation of Sweet Paste, a Chinese Traditional Seasoning, Using PCR-Denaturing Gradient Gel Electrophoresis

  • Mao, Ping;Hu, Yuanliang;Liao, Tingting;Wang, Zhaoting;Zhao, Shumiao;Liang, Yunxiang;Hu, Yongmei
    • Journal of Microbiology and Biotechnology
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    • 제27권4호
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    • pp.678-684
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    • 2017
  • The aim of this study was to elucidate the changes in the microbial community and biochemical properties of a traditional sweet paste during fermentation. PCR-denaturing gradient gel electrophoresis (DGGE) analysis showed that Aspergillus oryzae was the predominant species in the koji (the fungal mixture), and the majority of the fungi isolated belonged to two Zygosaccharomyces species in the mash. The bacterial DGGE profiles revealed the presence of Bacillus subtilis during fermentation, and Lactobacillus acidipiscis, Lactobacillus pubuzihii, Lactobacillus sp., Staphylococcus kloosi, and several uncultured bacteria were also detected in the mash after 14 days of main fermentation. Additionally, during main fermentation, amino-type nitrogen and total acid increased gradually to a maximum of $6.77{\pm}0.25g/kg$ and $19.10{\pm}0.58g/kg$ (30 days) respectively, and the concentration of reducing sugar increased to $337.41{\pm}3.99g/kg$ (7 days). The 180-day fermented sweet paste contained $261.46{\pm}19.49g/kg$ reducing sugar and its pH value remained at around 4.65. This study has used the PCR-DGGE technique to demonstrate the microbial community (including bacteria and fungi) in sweet paste and provides useful information (biochemical properties) about the assessment of the quality of sweet paste throughout fermentation.

PCR-DGGE Analysis of the Microbial Communities in Three Different Chinese "Baiyunbian" Liquor Fermentation Starters

  • Xiong, Xiaomao;Hu, Yuanliang;Yan, Nanfeng;Huang, Yingna;Peng, Nan;Liang, Yunxiang;Zhao, Shumiao
    • Journal of Microbiology and Biotechnology
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    • 제24권8호
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    • pp.1088-1095
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    • 2014
  • A systematic investigation was performed on the bacterial, Bacillus, fungal, and yeast communities of the three types of Daqu (mechanically prepared, manually prepared, and mixed prepared) used in Baiyunbian Company by reconditioning PCR-denaturing gradient gel electrophoresis (PCR-DGGE). The DGGE results showed that the microbes in the three types of Daqu were mainly thermotolerant and thermophilic microbes, and the most dominant bacterial species were Bacillus and Virgibacillus, followed by Lactobacillus and Trichococcus. Furthermore, the dominant fungi were found to be molds, such as Rasamsonia, Penicillium, Aspergillus, and Monascus, and the dominant yeasts were Saccharomyces cerevisiae, Saccharomycopsis fibuligera, Pichia anomala, and Debaryomyces hansenii. In general, the three types of Daqu showed slight differences in microbial communities, and the Shannon indexes (H') of the manually prepared and mechanically prepared Daqu were similar. The results suggest that mechanically prepared Daqu can replace manually prepared Daqu in liquor production, and this research provides useful information for liquor production and process improvement.

Senior Thai Fecal Microbiota Comparison Between Vegetarians and Non-Vegetarians Using PCR-DGGE and Real-Time PCR

  • Ruengsomwong, Supatjaree;Korenori, Yuki;Sakamoto, Naoshige;Wannissorn, Bhusita;Nakayama, Jiro;Nitisinprasert, Sunee
    • Journal of Microbiology and Biotechnology
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    • 제24권8호
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    • pp.1026-1033
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    • 2014
  • The fecal microbiotas were investigated in 13 healthy Thai subjects using polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Among the 186 DNA bands detected on the polyacrylamide gel, 37 bands were identified as representing 11 species: Bacteroides thetaiotaomicron, Bacteroides ovatus, Bacteroides uniformis, Bacteroides vulgatus, Clostridium colicanis, Eubacterium eligenes, E. rectale, Faecalibacterium prausnitzii, Megamonas funiformis, Prevotella copri, and Roseburia intestinalis, belonging mainly to the groups of Bacteroides, Prevotella, Clostridium, and F. prausnitzii. A dendrogram of the PCR-DGGE divided the subjects; vegetarians and non-vegetarians. The fecal microbiotas were also analyzed using a quantitative real-time PCR focused on Bacteroides, Bifidobacterium, Enterobacteriaceae, Clostrium coccoides-Eubacterium rectale, C. leptum, Lactobacillus, and Prevotella. The nonvegetarian and vegetarian subjects were found to have significant differences in the high abundance of the Bacteroides and Prevotella genera, respectively. No significant differences were found in the counts of Bifidabacterium, Enterobacteriaceae, C. coccoides-E. rectale group, C. leptum group, and Lactobacillus. Therefore, these findings on the microbiota of healthy Thais consuming different diets could provide helpful data for predicting the health of South East Asians with similar diets.

군부대와 주유소 주변에서 채취한 토양에서의 미생물 군집구조와 우점종의 파악 (Findings of Microbial Community Structure and Dominant Species in Soils Near Army Bases and Gas Stations)

  • 김재수
    • 대한환경공학회지
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    • 제32권3호
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    • pp.227-233
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    • 2010
  • 각종 오염지역의 오염토양의 시료를 채취해 DGGE를 이용한 미생물군집구조를 서로 비교함으로써 어떤 요인이 영향을 많이 미치는지를 알아보았다. 미생물군집의 유사도 비교에서 TPH 오염 말고는 대부분의 오염정도가 심하지 않아서 오염 특성보다는 토양시료가 채취된 지역에 따라 더 유사함을 보여주었고 TPH 오염도가 가장 높았던 시료에서만 지역적 특성을 넘어 유사도 차이를 보여 주었다. 이와 같이 오염요인이 너무 크지 않은 경우에는 미생물군집구조가 지역특성 즉 여러 다른 환경요인에 의해 더 크게 영향을 받는 것을 이 연구를 통해 알 수가 있었다. 모든 오염토양의 우점종(배양성 세균) 조사에서 firmicutes와 high GC Gram+에 속하는 세균들이 주로 발견되었고 유류오염이 심한 토양에서 이미 보고된 Arthrobacter, Bacillus, Methylobacterium, Clavibacter, Streptomyces, Nocardia와 아직 보고되지 않은 Leifsonia가 발견되었다.

Microbial Community Profiling in cis- and trans-Dichloroethene Enrichment Systems Using Denaturing Gradient Gel Electrophoresis

  • Olaniran, Ademola O.;Stafford, William H.L.;Cowan, Don A.;Pillay, Dorsamy;Pillay, Balakrishna
    • Journal of Microbiology and Biotechnology
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    • 제17권4호
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    • pp.560-570
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    • 2007
  • The effective and accurate assessment of the total microbial community diversity is one of the primary challenges in modem microbial ecology, especially for the detection and characterization of unculturable populations and populations with a low abundance. Accordingly, this study was undertaken to investigate the diversity of the microbial community during the biodegradation of cis- and trans-dichloroethenes in soil and wastewater enrichment cultures. Community profiling using PCR targeting the l6S rRNA gene and denaturing gradient gel electrophoresis (PCR-DGGE) revealed an alteration in the bacterial community profiles with time. Exposure to cis- and trans-dichloroethenes led to the disappearance of certain genospecies that were initially observed in the untreated samples. A cluster analysis of the bacterial DGGE community profiles at various sampling times during the degradation process indicated that the community profile became stable after day 10 of the enrichment. DNA sequencing and phylogenetic analysis of selected DGGE bands revealed that the genera Acinetobacter, Pseudomonas, Bacillus, Comamonas, and Arthrobacter, plus several other important uncultured bacterial phylotypes, dominated the enrichment cultures. Thus, the identified dominant phylotypes may play an important role in the degradation of cis- and trans-dichloroethenes.

Identification of Lactic Acid Bacteria in Galchi- and Myeolchi-Jeotgal by 16S rRNA Gene Sequencing, MALDI-TOF Mass Spectrometry, and PCR-DGGE

  • Lee, Yoonju;Cho, Youngjae;Kim, Eiseul;Kim, Hyun-Joong;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • 제28권7호
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    • pp.1112-1121
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    • 2018
  • Jeotgal is a Korean traditional fermented seafood with a high concentration of salt. In this study, we isolated lactic acid bacteria (LAB) from galchi (Trichiurus lepturus, hairtail) and myeolchi (Engraulis japonicas, anchovy) jeotgal on MRS agar and MRS agar containing 5% NaCl (MRS agar+5% NaCl), and identified them by using 16S rRNA gene sequencing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as culture-dependent methods. We also performed polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as a culture-independent method to identify bacterial communities. Five samples of galchi-jeotgal and seven samples of myeolchi-jeotgal were collected from different regions in Korea. A total of 327 and 395 colonies were isolated from the galchi- and myeolchi-jeotgal samples, respectively. 16S rRNA gene sequencing and MALDI-TOF MS revealed that the genus Pediococcus was predominant on MRS agar, and Tetragenococcus halophilus on MRS agar+5% NaCl. PCR-DGGE revealed that T. halophilus, Tetragenococcus muriaticus, and Lactobacillus sakei were predominant in both types of jeotgal. T. halophilus was detected in all samples. Even though the same species were identified by both culture-dependent and -independent methods, many species identified by the culture-dependent methods were not in the bacterial list identified by the culture-independent methods. The distribution of bacteria in galchi-jeotgal was more diverse than in myeolchi-jeotgal. The diverse LAB in galchi- and myeolchi-jeotgals can be further studied as candidates for starter cultures to produce fermented foods.

혐기성 수소생산 시 운전 pH 변화에 따른 미생물의 군집 변화 (Change of Microbial Communities in Fermentative Hydrogen Production at Difference Cultivation pHs)

  • 전윤선;이관용;조윤아;이태진
    • 대한환경공학회지
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    • 제30권12호
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    • pp.1239-1244
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    • 2008
  • 본 연구는 혐기성 발효에 의한 수소 생산 시 pH가 3에서 10까지 단계적으로 변화되는 조건에서 미생물의 군집 변화를 살펴보기 위해 PCR-DGGE를 실시하였다. 최대 수소생산 수율은 pH 5에서 1.8 mol $H_2$/mol substrate로 측정 되었으며, 각 pH에서 미생물의 성장량과 수소생산효율의 비례적 상관관계가 나타나지 않았다. 각 pH에서 채취된 미생물의 16S rDNA을 target으로 한 PCRDGGE를 수행한 결과, pH 조건에 따라 미생물의 군집 조성에 변화가 있음을 확인할 수 있었다. 미생물 종의 대부분은 Klebsiella 속으로 규명되었으며 Streptococcus 속과 Clostridium 속 미생물이 수소생산 효율에 많은 영향을 미치는 것으로 판단되었다.

사구식물 연관 세균 군집의 분자계통학적 연구 (Molecular Systematic Study of Bacterial Community Associated with Sand Dune Plants)

  • 도진옥;박성주;김승범
    • 환경생물
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    • 제25권4호
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    • pp.356-362
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    • 2007
  • 충남 태안군 일대 4곳의 해안사구 지역에서 자생하는 9종의 사구식물 연관 근권세균 다양성을 2003년 10월부터 2004년 3월까지의 기간 동안 3차례에 걸쳐 denaturing gradient gel electrophoresis (DGGE) 기법을 이용하여 조사하였다. 그 결과 한 밴드가 계속 모든 시료에서 우점하는 것으로 나타났으며, DNA 염기서열 분석에 의하여 Lysobacter enzymogenes와 가장 유사한 것으로 나타났다. 기타 주요 밴드들은 Pseudomonas와 Bacillus 속의 균주들로 동정되었다. L. enzymogenes가 9종의 식물 모두에서 지역이나 조사시기에 관계없이 우점적으로 나타난 것은 이전의 클론 분석을 통한 결과와 일치한다 (Lee et al. 2006a). Bacillus에 속한 밴드들이 모든 조사에서 출현하였으며, Pseudomonas 속 밴드들은 2003년 12월 조사에서 두드러졌다. DGGE 분석만으로는 Lysobacter가 사구식물에 가지는 중요성을 파악할 수는 없으나 건강한 개체에 지속적으로 발견되는 것으로 보아 Lysobacter의 존재는 식물에 긍정적인 영향을 가지는 것으로 사료된다.