• Journal of Microbiology and Biotechnology
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• v.23 no.5
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• pp.614-622
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• 2013

Daqu, a traditional fermentation starter, has been used to produce attractively flavored foods such as vinegar and Chinese liquor for thousands of years. Although Bacillus spp. are one of the dominant microorganisms in Daqu, more precise information is needed to reveal why and how Bacillus became dominant in Daqu, and next, to assess the impact of Bacillus sp. on Daqu and its derived products. We combined culture-dependent and culture-independent methods to study the ecology of Bacillus during Daqu incubation. Throughout the incubation, 67 presumptive Bacillus spp. isolates were obtained, 52 of which were confirmed by 16S rDNA sequencing. The identified organisms belonged to 8 Bacillus species: B. licheniformis, B. subtilis, B. amyloliquefaciens, B. cereus, B. circulans, B. megaterium, B. pumilus, and B. anthracis. A primer set specific for Bacillus and related genera was used in a selective PCR study, followed by a nested DGGE PCR targeting the V9 region of the 16S rDNA. Species identified from the PCR-DGGE fingerprints were related to B. licheniformis, B. subtilis, B. amyloliquefaciens, B. pumilus, B. benzoevorans, and B. foraminis. The predominant species was found to be B. licheniformis. Certain B. licheniformis strains exhibited potent antimicrobial activities. The greatest species diversity occurred at the Liangmei stage of Daqu incubation. To date, we lack sufficient knowledge of Bacillus distribution in Daqu. Elucidating the ecology of Bacillus during Daqu incubation would enable the impact of Bacillus on Daqu to be accessed, and the quality and stabilization of Daqu-derived products to be optimized.

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1128-1133
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• 2010

The diversity and abundance of ammonia-oxidizing bacteria (AOB) in activated sludge were compared using PCR-DGGE and real-time PCR assays. Activated sludge samples were collected from five different types of wastewater treatment plants (WWTPs) mainly treating textile, paper, food, and livestock wastewater or domestic sewage. The composition of total bacteria determined by PCR-DGGE was highly diverse between the samples, whereas the community of AOB was similar across all the investigated activated sludge. Total bacterial numbers and AOB numbers in the aerated mixed liquor were in the range of $1.8{\times}10^{10}$ to $3.8{\times}10^{12}$ and $1.7{\times}10^6$ to $2.7{\times}10^{10}$ copies/l, respectively. Activated sludge from livestock, textile, and sewage treating WWTPs contained relatively high amoA gene copies (more than $10^5$ copies/l), whereas activated sludge from food and paper WWTPs revealed a low number of the amoA gene (less than $10^3$ copies/l). The value of the amoA gene copy effectively showed the difference in composition of bacteria in different activated sludge samples and this was better than the measurement with the AOB 16S rRNA or total 16S rRNA gene. These results suggest that the quantification of the amoA gene can help monitor AOB and ammonia oxidation in WWTPs.

• Environmental Engineering Research
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• v.12 no.2
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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.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2004.09a
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• pp.175-178
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• 2004

Little study has been published specifically addressing the dynamics of nitrate reducing bacteria (NBR) during the bioremediation of nitrate-contaminated aquifer. In our previous study we successfully quantified fumarate-enhanced microbial nitrate reduction rate in a nitrate-contaminated aquifer by using a series of single-well push-pull tests (PPTs). In this study we analyzed the suspended population during PPTs. To monitor changes in the microbial community, PCR amplification of 16S rDNA genes and denaturing gradient gel electrophoresis (DGGE) were used to study the dynamics of the bacterial community in detail. Before the stimulation of NBR, the dominant DGGE bands obtained by PCR were affiliated with V-Proteobacteria consisting of Acinetobacter spp. and Pseudomonas fluorescens. However, as NBR biostimulation proceeded, the dominant patterns of DGGE bands changed, and they were affiliated with Azoarcus denitrificans Td-3 and Flavobacterium xanthum. Azoarcus denitrificans Td-3 is known to completely reduce nitrate to nitrogen gas. The series of single-well push-pull tests in this study should prove useful for conducting rapid, low-cost feasibility assessments for in situ denitrification and provide important information about which microorganisms play a key role in bioremediation of a nitrate contaminated aquifer.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.6
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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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• v.22 no.1
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• pp.1-12
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• 2012

The present study examines the effects of 30 mg/kg butachlor on the cyanobacterial diversity of rice fields in Eastern Uttar Pradesh and Western Bihar in India. A total of 40 samples were grouped into three classes [(i) acidic, (ii) neutral, and (iii) alkaline soils], based on physicochemical and principle component analyses. Acidic soils mainly harbored Westillopsis, Trichormus, Anabaenopsis, and unicellular cyanobacteria; whereas Nostoc, Anabaena, Calothrix, Tolypothrix, and Aulosira were found in neutral and alkaline soils. Molecular characterization using 16S rRNA PCR and DGGE revealed the presence of 13 different phylotypes of cyanobacteria in these samples. Butachlor treatment of the soil samples led to the disappearance of 5 and the emergence of 2 additional phylotypes. A total of 40 DGGE bands showed significant reproducible changes upon treatment with butachlor. Phylogenetic analyses divided the phylotypes into five major clusters exhibiting interesting links with soil pH. Aulosira, Anabaena, Trichormus, and Anabaenopsis were sensitive to butachlor treatment, whereas uncultured cyanobacteria, a chroococcalean member, Westillopsis, Nostoc, Calothrix, Tolypothrix, Rivularia, Gloeotrichia, Fischerella, Leptolyngbya, and Cylindrospermum, appeared to be tolerant against butachlor at their native soil pH. Butachlor-induced inhibition of nitrogen fixation was found to be 65% (maximum) and 33% (minimum) in the soil samples of pH 9.23 and 5.20, respectively. In conclusion, low butachlor doses may prove beneficial in paddy fields having a neutral to alkaline soil pH.

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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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.

• Journal of Korean Society of Environmental Engineers
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• v.36 no.1
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• pp.42-48
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• 2014

In this study, variations of the organic and nitrogenous compounds in wastewater were investigated in a single reactor with intermittent aeration. Over 90% of organic and nitrogen removals are accomplished with C/N ratio of 3 : 1 and 20/20 min of aeration/non-aeration period. Longer non-aeration period on the aeration/non-aeration cycle showed more stable nitrogen removal, showing various microbial community in the reactor. From PCR-DGGE analysis, it is conclusive that Dysgonomonas mossii strain Melo40, Eubacterium sp. oral clone JN088, Uncultured bacterium clone SPESB2_718, and Bacterium enrichment culture clone LE are related with the organics and nitrogen oxidation. Uncultured Acidobacteria bacterium clone AKYG487, Lactobacillus harbinensis strain FQ003, Erythrobacter litoralis strain Gi-3, Phytobacter diazotrophicus strain Ls8, and Mycobacterium sp. enrichment culture clone GE10037biofNNA are distinctly appeared under denitrification condition.

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.232-239
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• 2006

The microbial activity and bacterial community structure of activated sludge reactors, which treated free cyanide (FC), zinc-complexed cyanide (ZC), or nickel-complexed cyanide (NC), were studied. The three reactors (designated as re-FC, re-ZC, and re-NC) were operated for 50 days with a stepwise decrease of hydraulic retention time. In the re-FC and re-ZC reactors, FC or ZC was almost completely removed, whereas approximately 80-87% of NC was removed in re-NC. This result might be attributed to the high toxicity of nickel released after degradation of NC. In the batch test, the sludges taken from re-FC and re-ZC completely degraded FC, ZC, and NC, whereas the sludge from re-NC degraded only NC. Although re-FC and re-ZC showed similar properties in regard to cyanide degradation, denaturing gradient gel electrophoresis (DGGE) analysis of the 16S rRNA gene of the bacterial communities in the three reactors showed that bacterial community was specifically acclimated to each reactor. We found several bacterial sequences in DGGE bands that showed high similarity to known cyanide-degrading bacteria such as Klebsiella spp., Acidovorax spp., and Achromobacter xylosoxidans. Flocforming microorganism might also be one of the major microorganisms, since many sequences related to Zoogloea, Microbacterium, and phylum TM7 were detected in all the reactors.

• Journal of Korean Society on Water Environment
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• v.26 no.1
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• pp.10-18
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• 2010

Water quality and the distribution of ammonia oxidizing bacteria were characterized in constructed wetland of Shihwa lake. Both physico-chemical parameters and fecal indicator microorganisms including total coliforms, E.coli, Enterococcus spp. were measured. In addition, denaturant gradient gel electrophoresis (DGGE) was carried out after PCR amplification of amoA gene from input, output, and wetland sites of the Banwol, Donghwa, and Samhwa stream in Shihwa lake area. Physico-chemical parameters were in proper range for typical nitrifying bacteria to grow and perform their biological activities. Average concentrations of fecal indicator microorganisms of wetland samples were lower than those of input sites. These results suggested that microbial water quality improved by the process of constructed wetland. According to phylogenetic information obtained from DGGE from study sites, distribution of nitrifying bacteria from each of input, output, and wetland were generally distinctive one another. In addition, distribution of nitrifying bacteria between Banwol and Donghwa streams showed higher similarity (52.6%) than this of Samhwa stream (15.2%). These results indicated that characteristics of ammonia oxidizing bacteria in Samhwa were unique in comparison with those of Banwol and Donghwa stream.