• Journal of Korean Society of Environmental Engineers
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• v.34 no.5
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• pp.304-311
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• 2012

Chlorinated ethylenes such as perchloroethylene (PCE) and trichloroethylene (TCE) are widely used as industrial solvents and degreasing agents. Because of improper handling, these highly toxic chlorinated ethylenes have been often detected from contaminated soils and groundwater. Biological PCE dechlorination activities were tested in bacterial cultures inoculated with 10 different environmental samples from sediments, sludges, soils, and groundwater. Of these, the sediment using culture (SE 2) was selected and used for establishing an efficient PCE dechlorinating enrichment culture since it showed the highest activity of dechlorination. The cathode chamber of bioelectrochemical system (BES) was inoculated with the enrichment culture and the system with a cathode polarized at -500 mV (Vs Ag/AgCl) was operated under fed-batch mode. PCE was dechlorinated to ethylene via TCE, cis-dichloroethylene, and vinyl chloride. Microbial community analysis with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) showed that the microbial community in the enrichment culture was significantly changed during the bio-electrochemical PCE dechlorination in the BES. The communities of suspended-growth bacteria and attached-growth bacteria on the cathode surface are also quite different from each other, indicating that there were some differences in their mechanisms receiving electrons from electrode for PCE dechlorination. Further detailed research to investigate electron transfer mechanism would make the bioelctrochemical dechlorination technique greatly useful for bioremediation of soil and groundwater contaminated with chlorinated ethylenes.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1330-1335
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• 2012

Foot and mouth disease (FMD) is one of the acute infectious diseases in hoofed and even-toed mammals, including pigs, and it occurs via acute infection by Aphthovirus. When FMD is suspected, animals around the location of origin are typically slaughtered and buried. Other methods such as rendering, composting, and incineration have not been verified in practice in Korea. After the FMD incident, the regular monitoring of the microbial community is required, as microorganisms greatly modify the characteristics of the ecosystem in which they live. This is the result of their metabolic activities causing chemical changes to take place in the surrounding environment. In this study, we investigated changes in the microbial community during a 24 week period with DNA extracts from leachate, formed by the decomposition of buried pigs at a laboratory test site, using denaturing gradient gel electrophoresis (DGGE) with a genomic DNA. Our results revealed that Bacteroides coprosuis, which is common in pig excreta, and Sporanaerobacter acetigenes, which is a sulfur-reduced microbe, were continuously observed. During the early stages (0~2 weeks) of tissue decomposition, Clostridium cochlearium, Fusobacterium ulcerans, and Fusobacterium sp., which are involved in skin decomposition, were also observed. In addition, various microbes such as Turicibacter sanguinis, Clostridium haemolyticum, Bacteroides propionicifaciens, and Comamonas sp. were seen during the later stages (16~24 weeks). In particular, the number of existing microbial species gradually increased during the early stages, including the exponential phase, decreased during the middle stages, and then increased again during the later stages. Therefore, these results indicate that the decomposition of pigs continues for a long period of time and leachate is created continuously during this process. It is known that leachate can easily flow into the neighboring environment, so a long-term management plan is needed in burial locations for FMD-infected animals.

• Journal of Soil and Groundwater Environment
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• v.17 no.5
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• pp.49-55
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• 2012

A 1.28 L-batch reactor and continuous-flow stirred tank reactor (CFSTR) fed with formate and trichloroethene (TCE) were operated for 120 days and 56 days, respectively, to study the effect of formate as electron donor on anaerobic reductive dechlorination (ARD) of TCE to cis-1,2-dichloroethylene (c-DCE), vinyl chloride (VC), and ethylene (ETH). In batch reactor, injected 60 ${\mu}mol$ TCE was completely degraded in the presence of 20% hydrogen gas ($H_2$) in less than 8 days by anaerobic dechlorination mixed-culture (300 mg-soluble protein), Evanite Culture with ability to completely degrade tetrachloroethene (PCE) and -TCE to ETH under anaerobic conditions. Once the formate was used as electron donor instead of hydrogen gas in batch or chemostat system, the TCE-dechlorination rate decreased and acetate production rate increased. It indicates that the concentration of hydrogen produced in both systems is possibly more close to threshold for homoacetogenesis process. Soluble protein concentration of Evanite culture during the batch test increased from 300 mg to 688 mg for 120 days. Through the protein monitoring, we confirmed an increase of microbial population during the reactor operation. In CFSTR test, TCE was fed continuously at 9.9 ppm (75.38 ${\mu}mol/L$) and the influent formate feed concentration increased stepwise from 1.3 mmol/L to 14.3 mmol/L. Injected TCE was accumulated at 18 days of HRT, but TCE was completely degraded at 36 days of HRT without accumulation of the injected-TCE during the left of experiment period, getting $H_2$ from fermentative hydrogen production of injected formate. Although c-DCE was also accumulated for 23 days after beginning of CFSTR operation, it reached steady-state in the presence of excessive formate. We also evaluated microbial dynamic of the culture at different chemical state in the reactor by DGGE (denaturing gradient gel electrophoresis).

• Korean Journal of Environmental Agriculture
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• v.38 no.2
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• pp.76-82
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• 2019

BACKGROUND: For sustainable agriculture, there are various agricultural practices including low input. Over the last few decades high input of chemical fertilizer and compounds results in environmental pollution and deterioration of soil fertility. Soybean (Glycine max L.) is well known eco-friendly crop due to their symbionts. Soybean has a relationship with nitrogen fixation bacteria called rhizobia. In this research work, we investigated effects of soybean cultivation on soil microorganism activities. METHODS AND RESULTS: Experiments were conducted in pots and potato cultivation was used as reference. Soil chemical properties were analyzed considering soil nutrient over cropping period. For the soil microbial community analysis, dehydrogenase activity analysis (DHA) analyzed along with denaturing gradient gel electrophoresis. The results showed that higher soil organic matter in the soybean cultivation soil than in the potato cultivation soil. Available $P_2O_5$ concentration increased gradually in both pots but showed higher value in the potato cultivation soil. DHA value implying microbial activities showed higher value in the soybean cultivation soil over all cropping period. CONCLUSION: The cause of high microbial activity in the soybean cultivation soil was considered to the effects of some specific microorganisms related to soybean cultivation. Therefore, the availability of soybean cultivation for sustainable agriculture should be encouraged in terms of microorganism community activity in soil.

• 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.

• 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.

• 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.

• Microbiology and Biotechnology Letters
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• v.41 no.3
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• pp.292-299
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• 2013

In this study, we analyzed the changes in fungal populations of red-pepper fields employing eco-friendly farming methods, such as microbial agents and crop rotation, by using polymerase chain reactions coupled with denaturing gradient gel electrophoresis (PCR-DGGE). Primer specific for fungi were used to determine the contribution of domains to the microbial community. Analysis of planted and non-planted soil samples applying PCR-DGGE technology offered evaluation of long-term patterns in fungal species richness. To evaluate the stability of DGGE patterns from different soils, comparison of planted and non-planted soil samples were compared using PCR-DGGE. The number of DNA fragments obtained from all planted soil samples by DGGE separation was far greater (14 to 15 bands) than that of the non-planted soil samples (3 to 4 bands). In addition, 14 bands were observed from crop continuation soil treated with agrochemicals and 18 bands from crop rotation soil treated with microbial agents. The PCR-DGGE analysis suggests that the use of crop rotation and microbial agents benefits the fungal community more than crop continuation using agrochemicals. These results indicate that crop rotation with microbial agents was better able to support beneficial organisms, enable more effective biological control and maintain a healthier balance of nutrients, organic matter and microorganisms.

• Food Engineering Progress
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• v.15 no.4
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• pp.370-375
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• 2011

In this study the polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens in fresh originally grown vegetables. A universal primer (341GCf/534r) was selected for its ability to amplify the V3 region of 16S-rRNA genes in their target pathogens (Salmonella typhimurium, Pseudomonas fluorescens, Bacillus cereus, Listeria monoytogenes, Staphyloocus aureus, E. coli). The 194 bp fragments in PCR were successfully duplicated as expected. The amplified fragments of the same size from six different pathogens also showed good separation upon DGGE. The detection limit of PCR-DGGE for six pathogens in fresh-cut lettuces were over $10^{5}$ CFU/g when sampled by stomaching. However, when the sampling method was changed from stomaching to shaking, the detection limit of six pathogens in organic vegetables was shown to increase by over $10^{1}$ CFU/g, but only those of B. cereus were over $10^{3}$ CFU/g. Therefore, PCR-DGGE was shown to be a reliable method for the detection of pathogens in fresh-cut vegetables.

• Proceedings of the Korean Society of Organic Agriculture Conference
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• 2009.12a
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• pp.309-309
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• 2009

국내외 환경문제와 관련하여 농업환경보전 및 안전한 먹거리에 대한 소비자의 요구가 커지면서 친환경농업에 대한 관심이 높이지고 있다. 친환경농업은 화학비료와 농약의 사용을 양분종합관리 및 병해충종합관리등을 통해 저투입하는 농법과 화학비료와 농약의 대체 농자재를 사용하는 유기농법으로 나누어져 있다. 대체 농자재는 목초액, 키토산 등 시판되는 자재를 비롯하여 천혜녹즙, 한방영양제, 아미노산액비 등과 같이 농가를 중심으로 자가 제조되어 활용되고 있는 것이 특징이다. 그러나 대부분 효능이 제대로 검증되지 않고 임의로 사용되고 있기 때문에 본 연구는 농가 자가제조 유기농액비의 미생물학적 특성을 구명하여 유기농 액비의 농업적 활용 제고를 위한 표준화를 확립하기 위해 수행하였다. 농가자가 제조 유기농액비는 성환과 제주에서 5점을 수집하여 액비의 화학성과 미생물상을 비교하였다. 액비 제조시 종균으로 사용되고 있는 부엽토를 5지역에서 채취하여 세균군집의 차이를 보기위해 denaturing gradient gel electrophoresis (DGGE)를 수행하여 분석하였다. 주요 결과를 요약하면 수집된 액비의 화학성 분석결과, pH는 3.3~4.3로 강한 산성을 나타났으며 EC는 $32.8dS{\cdot}m^{-1}{\sim}552.33dS{\cdot}m^{-1}$ 이었다. 수집된 액비내 미생물상을 조사한 결과, 세균과 방선균은 존재하였지만 사상균은 검출되지 않았다 또한 성환에서 수집된 액비로부터 세균을 분리하여 16S rRNA 염기서열을 분석한 결과, 대부분 Bacillus속으로 95%이상의 높은 유사성을 갖는 것으로 나타났다. 액비 발효 시 종균으로 사용되고 있는 부엽토의 미생물상을 조사한 결과, 지역에 따라 세균은 0.8~$110{\times}10^5cfu{\cdot}g^{-1}$, 방선균은 1.0~$10.9{\times}10^4cfu{\cdot}g^{-1}$, 그리고 사상균은 2.6~$64{\times}10^3cfu{\cdot}g^{-1}$의 균수를 보였다. 부엽토간의 세균군집의 차이를 비교하기 위해 DGGE를 수행한 결과, gel 상에서 다양한 위치의 밴드를 확인할 수 있으며, 부엽토별로 공통적 및 특이적 밴드를 확인할 수 있었다.