• Title/Summary/Keyword: DGGE

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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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    • v.24 no.6
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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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    • v.27 no.4
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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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    • v.24 no.8
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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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    • v.24 no.8
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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.

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

  • Kim, Jai-Soo
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.3
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    • pp.227-233
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    • 2010
  • This study examined microbial community structures (MCSs) according to environmental factors through DGGE analysis and comparison in various soils collected from near army bases and gas stations. As a result, the similarities based on DGGE band profiles showed the closer relationship in regional properties than in pollution characteristics, probably due to the degree of weak contamination. The highly contaminated samples with oil revealed low MCS similarities with others in the same region and very low with all the other samples in the other regions. Thus the microbial community structure would more be affected by region-based natural factors than by contamination factors in case of minor pollution. All the dominant culturable bacterial species were involved in firmicutes or high GC Gram+ in a major portion of soil samples and the highly oil-contaminated samples contained Arthrobacter, Bacillus, Methylobacterium, Clavibacter, Streptomyces and Nocardia as reported genera, and Leifsonia as a unreported genus.

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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    • v.17 no.4
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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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    • v.28 no.7
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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.

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

  • Jun, Yoon-Sun;Lee, Kwan-Yong;Cho, Yoon-A;Lee, Tae-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.12
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    • pp.1239-1244
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    • 2008
  • In this study, PCR-DGGE was conducted to investigate the variations of microbial community according to pH conditions from pH 3 to pH 10 during anaerobic fermentation process of hydrogen production. Maximum hydrogen yield was 1.8 mol $H_2$/mol substrate at pH 5. The microbial growth rate was not proportional to the hydrogen production rate at each pH. Variations of microbial community was observed at each condition from PCR-DGGE experiment of 16s rDNA. Klebsiella was main species of the microbial community. Streptococcus and Clostridium were mainly contributed for hydrogen production.

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

  • Do, Jin-Ok;Park, Seong-Joo;Kim, Seung-Bum
    • Korean Journal of Environmental Biology
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    • v.25 no.4
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    • pp.356-362
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    • 2007
  • The rhizobacterial diversity associated with 9 native plant species inhabiting coastal sand dunes in Tae-an area, Chungnam Province, was studied using the denaturing gradient gel electrophoresis (DGGE) fingerprinting analysis over three times from October 2003 to March 2004. One dominant band commonly occurred in all of the rhizosphere samples, which was identified as that of Lysobacter enzymogenes. The other common bands included those derived from species of Pseudomonas and Bacillus. It was notable that L. enzymogenes was dominant in all of the 9 plant species and such dominance was consistent throughout the whole sampling period, which confirms the previous study by Lee et al. (2006a). The Bacillus bands were detected in all of the three samplings, and those of Pseudomonas were notable in the samples of December 2003. By the DGGE analysis alone, the significance of Lysobacter to the sand dune plants is not clear. However, considering their presence in healthy plants and the dominance in all plant species, Lysobacter may have positive roles in the survival or growth of the plants in sand dune area.