• 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.

• Journal of Microbiology and Biotechnology
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• 제23권1호
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• pp.76-84
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• 2013

A polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique followed by sequencing of the 16S rDNA fragments eluted from the bands of interest on denaturing gradient gels was used to monitor changes in the bacterial microflora of two commercial kimchi, salted cabbage, and ingredient mix samples during 30 days of fermentation at $4^{\circ}C$ and $10^{\circ}C$. Leuconostoc (Lc.) was the dominant lactic acid bacteria (LAB) over Lactobacillus (Lb.) species at $4^{\circ}C$. Weissella confusa was detected in the ingredient mix and also in kimchi samples throughout fermentation in both samples at $4^{\circ}C$ and $10^{\circ}C$. Lc. gelidum was detected as the dominant LAB at $4^{\circ}C$ in both samples. The temperature affected the LAB profile of kimchi by varing the pH, which was primarily caused by the temperature-dependent competition among different LAB species in kimchi. At $4^{\circ}C$, the sample variations in pH and titratable acidity were more conspicuous owing to the delayed growth of LAB. Temperature affected only initial decreases in pH and initial increases in viable cell counts, but affected both the initial increases and final values of titratable acidity. The initial microflora in the kimchi sample was probably determined by the microflora of the ingredient mix, not by that of the salted cabbage. The microbial distributions in the samples used in this study resembled across the different kimchi samples and the different fermentation temperatures as the numbers of LAB increased and titratable acidity decreased.

• 한국수산과학회지
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• 제40권6호
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• pp.356-361
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• 2007

Recently, the development of various culture-independent identification techniques for environmental microbes has greatly enhanced our knowledge of microbial diversity. In particular, denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments, generated using the polymerase chain reaction (PCR) is frequently used to examine the diversity of environmental bacterial populations. This method consists of direct extraction of the environmental DNA, amplification of the 200-600 bp 16S rDNA fragments with universal primers, and separation of the fragments according to their melting point on a denaturing gradient gel. In this study, we investigated the seaside microbial community in coastal areas of Busan, Korea, using culture-independent techniques. First, marine genomic DNA was extracted from seawater samples collected at Songjeong, Gwangahn, and Songdo Beaches. Then, PCR was used to amplify the bacterial 16S rDNA using universal primers, and DGGE was used to separate the amplified 500 bp 16S rDNA fragments. Finally, the tested 16S rDNA genes were further analyzed by sequencing. Based on these experiments, we found that DGGE analysis clearly showed variation among the regional groups. It can be used to monitor rapid changes in the bacterial diversity of various environments. In addition, the sequence analysis indicated the existence of many unculturable bacteria, in addition to Arcobacter, Pseudoaltermonas, and Vibrio species.

• 상하수도학회지
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• 제20권3호
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• pp.461-467
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• 2006

The seasonal change in attached algae and microbial community structure at sedimentation basin of water treatment plant was investigated by using quinone profiles and denaturing gel gradient electrophoresis (DGGE). The photosynthetic bacteria and algae contains PQ-9 and VK-1 as major quinone are major component of the total quinone fraction in attached algae and microorganisms on sedimentation basin trough. The microorganisms containing menaquinones appear to be sensitivity to the change in temperature than those containing ubiquinones. The plot of the mole fraction of dominant quinone species ($f_d$) to the DQ values showed higher sensitivity to the seasonal change in the microbial community structure. The results indicated that quinone and DGGE are useful tool for the evaluation of the changes in the microbial community structure.

• 생명과학회지
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• 제22권1호
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• pp.110-116
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• 2012

누룩은 탁주와 약주의 제조를 위한 당화효소와 알코올발효를 위한 미생물의 공급원으로서 제품의 맛과 품질을 결정하는 중요한 역할을 한다. 본 연구에서는 산성누룩의 세균과 진균의 다양성을 조사하기 위해 순수분리 종과 16S 및 28S rRNA gene를 대상으로 한 PCR-DGGE를 이용한 분석을 수행하였다. 누룩 내 세균의 수는 $2.7{\times}10^9$ CFU/g이었으며 순수분리와 PCR-DGGE 분석에서 우점종은 Kocuria spp., Pantoea spp., Lactobacillus spp., Pediococcous spp., Weissella spp., Staphylococcus spp. 그 외 endophytic bacterium, uncultured gamma-proteobacteria, uncultured Cyanobacteria와 Actinobacteria였다. PCR-DGGE profile에서 주된 우점종은 Pediococcous pentosaceus와 uncultured Cyanobacteria 이었다. 누룩 내 진균의 수는 $3.5{\times}10^8$ CFU/g이었으며 순수분리와 PCR-DGGE 분석에서 우점종은 Trichomonascus spp., Pichia spp., Torulaspora spp., Wickerhamomyces spp., Sacharomycopsis spp., Lichtheimia spp., Mucor spp., Rhizopus spp., Aspergillus spp., Cladosporium spp.였다. PCR-DGGE profile에서 주된 우점종은 Pichia kudriavzevii와 Aspergillus oryzae이었다. PCR-DGGE 기술은 본 연구에서 누룩의 미생물군집을 평가하기 위해 처음으로 사용되었으며 미생물 다양성을 설명하는 데 효과적임을 입증하였다.

• 미생물학회지
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• 제41권4호
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• pp.281-286
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• 2005

울산, 여수 등 공단지역의 토양, 하천의 저니, 해양의 준설토 둥을 이용하여 난분해성 염소화합물인 PCE (perchloroethylene)및 TCE (trichloroethylene)의 혐기성 탈염소화에 관련하는 미생물을 탐색하고 이들의 탈염소화 효율을 조사하였다. 혐기성 상호대사에 의한 탈염소화 효율을 조사하기 위해 전자공여체로 acetate를 사용하여 혐기성 회분식 실험을 실시하였으며, 이와 병행하여 분자생물학적인 기법인 16S rDNA의 PCR-Double Gradient DGGE (DG-DGGE)를 이용하여 미생물의 군집을 분석하였다. 그 결과 울산 태화강 및 여수 하남천의 저니를 접종한 경우 PCE는 $70\%,\;65\%$, TCE는 $50\%,\;45\%$의 높은 탈염소화 효율을 나타내었다. 또한 16S rDNA의 PCR을 이용한 DG-DGGE로 미생물 군집을 분석한 결과, 탈염소화 효율이 높은 지역의 저니에는 Desulfovibrio sup.의 미생물이 주로 존재함을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제17권1호
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• pp.67-73
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• 2007

The present study compared the microbial diversity and activity during the application of various bioremediation processes to crude oil-contaminated soil. Five different treatments, including natural attenuation (NA), biostimulation (BS), biosurfactant addition (BE), bioaugmentation (BA), and a combined treatment (CT) of biostimulation, biosurfactant addition, and bioaugmentation, were used to analyze the degradation rate and microbial communities. After 120 days, the level of remaining hydrocarbons after all the treatments was similar, however, the highest rate (k) of total petroleum hydrocarbon (TPH) degradation was observed with the CT treatment (P<0.05). The total bacterial counts increased during the first 2 weeks with all the treatments, and then remained stable. The bacterial communities and alkane monooxygenase gene fragment, alkB, were compared by denaturing gradient gel electrophoresis (DGGE). The DGGE analyses of the BA and CT treatments, which included Nocardia sp. H17-1, revealed a simple dominant population structure, compared with the other treatments. The Shannon-Weaver diversity index (H') and Simpson dominance index (D), calculated from the DGGE profiles using 16S rDNA, showed considerable qualitative differences in the community structure before and after the bioremediation treatment as well as between treatment conditions.

• Environmental Engineering Research
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• 제12권2호
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• pp.37-45
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• 2007

Phytoremediation has been used effectively for the biodegradation of oil-based contaminants, including diesel, by the stimulation of soil microbes near plant roots (rhizosphere). However, the technique has rarely been assessed for itsinfluence on soil microbial properties such as population, community structure, and diversity. In this study, the removal efficiency and characteristics of rhizobacteria for phytoremediation of diesel-contaminated soils were assessed using barnyard grass (Echinochloa crusgalli). The concentration of spiked diesel for treatments was around $6000\;mg\;kg^{-1}$. Diesel removal efficiencies reached 100% in rhizosphere soils, 76% in planted bulk soils, and 62% in unplanted bulk soils after 3weeks stabilization and 2 months growth(control, no microbial activity: 32%). The highest populations of culturable soil bacteria ($5.89{\times}10^8$ per g soil) and culturable hydrocarbon-degraders($5.65{\times}10^6$ per g soil) were found in diesel-contaminated rhizosphere soil, also yielding the highest microbial dehydrogenase. This suggests that the populations of soil bacteria, including hydrocarbon-degraders, were significantly increased by a synergistic rhizosphere + diesel effect. The diesel treatment alone resulted in negative population growth. In addition, we investigated the bacterial community structures of each soil sample based on DGGE (Denaturing Gel Gradient Electrophoresis) band patterns. Bacterial community structure was most influenced by the presence of diesel contamination (76.92% dissimilarity to the control) and by a diesel + rhizosphere treatment (65.62% dissimilarity), and least influenced by the rhizosphere treatment alone (48.15% dissimilarity). Based on the number of distinct DGGE bands, the bacterial diversity decreased with diesel treatment, but kept constant in the rhizosphere treatment. The rhizosphere thus positively influenced bacterial population density in diesel-contaminated soil, resulting in high removal efficiency of diesel.

• Asian-Australasian Journal of Animal Sciences
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• 제27권6호
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• pp.898-906
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• 2014

The objective of this study was to investigate differences in the faecal microbial composition among Lantang, Bama, Erhualian, Meishan, Xiaomeishan, Duroc, Landrace, and Yorkshire sows and to explore the possible link of the pig breed with the gut microbial community. Among the sows, the Meishan, Landrace, Duroc, and Yorkshire sows were from the same breeding farm with the same feed. Fresh faeces were collected from three sows of each purebred breed for microbiota analysis and volatile fatty acid (VFA) determination. Denaturing gradient gel electrophoresis (DGGE) analysis revealed that samples from Bama, Erhualian, and Xiaomeishan sows, which from different farms, were generally grouped in one cluster, with similarity higher than 67.2%, and those from Duroc, Landrace, and Yorkshire sows were grouped in another cluster. Principal component analysis of the DGGE profile showed that samples from the foreign breeds and the samples from the Chinese indigenous breeds were scattered in two different groups, irrespective of the farm origin. Faecal VFA concentrations were significantly affected by the pig breed. The proportion of acetate was higher in the Bama sows than in the other breeds. The real-time PCR analysis showed that 16S rRNA gene copies of total bacteria, Firmicutes and Bacteroidetes were significantly higher in the Bama sows compared to Xiaomeishan and Duroc sows. Both Meishan and Erhualian sows had higher numbers of total bacteria, Firmicutes, Bacteroidetes and sulphate-reducing bacteria as compared to Duroc sows. The results suggest that the pig breed affects the composition of gut microbiota. The microbial composition is different with different breeds, especially between overseas breeds (lean type) and Chinese breeds (relatively obese type).

• Journal of Microbiology and Biotechnology
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• 제22권1호
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• pp.126-132
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• 2012

A cellulose-degrading composite microbial system containing a mixture of microbes was previously shown to demonstrate a high straw-degrading capacity. To estimate its potential utilization as an inoculant to accelerate straw biodegradation after returning straw to the field, two cellulose-degrading composite microbial systems named ADS3 and WSD5 were inoculated into wheat straw-amended soil in the laboratory. The microbial survival of the inoculant was confirmed by a denaturing gradient gel electrophoresis (DGGE) analysis, whereas the enhancement of straw degradation in soil was assessed by measuring the mineralization of the soil organic matter and the soil cellulase activity. The results indicated that most of the DGGE bands from ADS3 were detected after inoculation into straw-amended autoclaved soil, yet only certain bands from ADS3 and WSD5 were detected after inoculation into straw-amended non-autoclaved soil during five weeks of incubation; some bands were detected during the first two weeks after inoculation, and then disappeared in later stages. Organic matter mineralization was significantly higher in the soil inoculants ADS3 and WSD5 than in the uninoculated controls during the first week, yet the enhanced degradation did not persist during the subsequent incubation. Similar to the increase in soil organic matter, the cellulase activity also increased during the first week in the ADS3 and WSD5 treatments, yet decreased during the remainder of the incubation period. Thus, it was concluded that, although the survival and performance of the two inoculants did not persist in the soil, a significant enhancement of degradation was present during the early stage of incubation.