• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1473-1483
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• 2014

Reports of herbicide resistance events are proliferating worldwide, leading to new cultivation strategies using combinations of pre-emergence and post-emergence herbicides. We analyzed the impact during a one-year cultivation cycle of several herbicide combinations on the rhizobacterial community of glyphosate-tolerant Bt-maize and compared them to those of the untreated or glyphosate-treated soils. Samples were analyzed using pyrosequencing of the V6 hypervariable region of the 16S rRNA gene. The sequences obtained were subjected to taxonomic, taxonomy-independent, and phylogeny-based diversity studies, followed by a statistical analysis using principal components analysis and hierarchical clustering with jackknife statistical validation. The resilience of the microbial communities was analyzed by comparing their relative composition at the end of the cultivation cycle. The bacterial communites from soil subjected to a combined treatment with mesotrione plus s-metolachlor followed by glyphosate were not statistically different from those treated with glyphosate or the untreated ones. The use of acetochlor plus terbuthylazine followed by glyphosate, and the use of aclonifen plus isoxaflutole followed by mesotrione clearly affected the resilience of their corresponding bacterial communities. The treatment with pethoxamid followed by glyphosate resulted in an intermediate effect. The use of glyphosate alone seems to be the less aggressive one for bacterial communities. Should a combined treatment be needed, the combination of mesotrione and s-metolachlor shows the next best final resilience. Our results show the relevance of comparative rhizobacterial community studies when novel combined herbicide treatments are deemed necessary to control weed growth.

• Journal of Microbiology and Biotechnology
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• 제23권10호
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• pp.1472-1477
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• 2013

Microalgae have been regarded as a natural resource for sustainable materials and fuels, as well as for removal of nutrients and micropollutants from wastewater, and their interaction with bacteria in wastewater is a critical factor to consider because of the microbial diversity and complexity in a variety of wastewater conditions. Despite their importance, very little is known about the ecological interactions between algae and bacteria in a wastewater environment. In this study, we characterized the wastewater bacterial community in response to the growth of a Selenastrum gracile UTEX 325 population in a real municipal wastewater environment. The Roche 454 GS-FLX Titanium pyrosequencing technique was used for indepth analysis of amplicons of 16S rRNA genes from different conditions in each reactor, with and without the algal population. The algal growth reduced the bacterial diversity and affected the bacterial community structure in the wastewater. The following in-depth analysis of the deep-sequenced amplicons showed that the algal growth selectively stimulated Sphingobacteria class members, especially the Sediminibacterium genus population, in the municipal wastewater environment.

• Journal of Microbiology and Biotechnology
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• 제21권3호
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• pp.293-298
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• 2011

Nitrifying bacterial community structures of suspended and attached biomasses in a full-scale integrated fixed-film activated sludge process were investigated by analyzing 16S rRNA gene sequences obtained from pyrosequencing. The suspended biomass had a higher number of ammonia-oxidizing bacterial sequences (0.8% of total sequences) than the attached biomass (0.07%), although most of the sequences were within the Nitrosomonas oligotropha lineage in both biomasses. Nitrospira-like nitrite-oxidizing bacterial sequences were retrieved in the suspended biomass (0.06%), not in the attached biomass, whereas the existence of Nitrobacter-like sequences was not evident. The suspended biomass had higher nitrification activity (1.13 mg N/TSS/h) than the attached biomass (0.07 mg N/TSS/h). Overall, the results made it possible to conclude the importance of the suspended biomass, rather than the attached biomass, in nitrification in the wastewater treatment process studied.

• 대한수의학회지
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• 제57권3호
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• pp.159-167
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• 2017

The microbial community is known to have a key role during the rearing period of broilers. In this study, gut microbial composition and diversity were examined to evaluate the relationships between these factors and broiler growth performance. By applying 454-pyrosequencing of the V1-V3 regions of bacterial 16S rRNA genes, six fecal samples from four- and 28-day-old chickens from three broiler farms and 24 intestinal samples of broilers with heavy and light body weights were analyzed. Microbial composition assessment revealed Firmicutes to be the most prevalent phylum at farm A, while Proteobacteria were predominant at farms B and C. Fecal microbial richness and diversity indices gradually increased from four to 28 days at all three farms. Microbial diversity assessment revealed that small intestine microbial diversity was lower in heavy birds than in light birds. In light birds, the Firmicutes proportion was lower than that in heavy birds. In conclusion, each broiler farm revealed a specific microbial profile which varied with the age of the birds. The microbial communities appeared to affect growth performance; therefore, gut microbial profiles can be utilized to monitor growth performance at broiler farms.

• Journal of Microbiology and Biotechnology
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• 제26권12호
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• pp.2141-2147
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• 2016

As one of the most complex human-associated microbial habitats, the oral cavity harbors hundreds of bacteria. Halitosis is a prevalent oral condition that is typically caused by bacteria. The aim of this study was to analyze the microbial communities and predict functional profiles in supragingival plaque from healthy individuals and those with halitosis. Ten preschool children were enrolled in this study; five with halitosis and five without. Supragingival plaque was isolated from each participant and 16S rRNA gene pyrosequencing was used to identify the microbes present. Samples were primarily composed of Actinobacteria, Bacteroidetes, Proteobacteria, Firmicutes, Fusobacteria, and Candidate phylum TM7. The ${\alpha}$ and ${\beta}$ diversity indices did not differ between healthy and halitosis subjects. Fifteen operational taxonomic units (OTUs) were identified with significantly different relative abundances between healthy and halitosis plaques, and included the phylotypes of Prevotella sp., Leptotrichia sp., Actinomyces sp., Porphyromonas sp., Selenomonas sp., Selenomonas noxia, and Capnocytophaga ochracea. We suggest that these OTUs are candidate halitosis-associated pathogens. Functional profiles were predicted using PICRUSt, and nine level-3 KEGG Orthology groups were significantly different. Hub modules of co-occurrence networks implied that microbes in halitosis dental plaque were more highly conserved than microbes of healthy individuals' plaque. Collectively, our data provide a background for the oral microbiota associated with halitosis from supragingival plaque, and help explain the etiology of halitosis.

• Journal of Microbiology and Biotechnology
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• 제24권7호
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• pp.888-897
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• 2014

A bacterial community analysis of the gut of Tenebrio molitor larvae was performed using pyrosequencing of the 16S rRNA gene. A predominance of genus Spiroplasma species in phylum Tenericutes was observed in the gut samples, but there was variation found in the community composition between T. molitor individuals. The gut bacteria community structure was not significantly affected by the presence of antibiotics or by the exposure of T. molitor larvae to a highly diverse soil bacteria community. A negative relationship was identified between bacterial diversity and ampicillin concentration; however, no negative relationship was identified with the addition of kanamycin. Ampicillin treatment resulted in a reduction in the bacterial community size, estimated using the 16S rRNA gene copy number. A detailed phylogenetic analysis indicated that the Spiroplasma-associated sequences originating from the T. molitor larvae were distinct from previously identified Spiroplasma type species, implying the presence of novel Spiroplasma species. Some Spiroplasma species are known to be insect pathogens; however, the T. molitor larvae did not experience any harmful effects arising from the presence of Spiroplasma species, indicating that Spiroplasma in the gut of T. molitor larvae do not act as a pathogen to the host. A comparison with the bacterial communities found in other insects (Apis and Solenopsis) showed that the Spiroplasma species found in this study were specific to T. molitor.

• Journal of Microbiology and Biotechnology
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• 제25권5호
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• pp.709-717
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• 2015

Airborne bacteria from hog farms may have detrimental impacts on human health, particularly in terms of antibiotic resistance and pathogen zoonosis. Despite human health risks, very little is known about the composition and diversity of airborne bacteria from hog farms and hog-related spray fields. We used pyrosequencing analysis of 16S rRNA genes to compare airborne bacterial communities in a North Carolina hog farm and lagoon spray field. In addition, we isolated and identified antibiotic-resistant bacteria from both air samples. Based on 16S rRNA gene pyrosequence analysis, Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria were the dominant phyla in airborne bacterial communities from both hog farm and spray field sites. Within the Firmicutes genera, Clostridium spp. were more abundant in the hog farm, whereas Staphylococcus spp. were higher in the spray field. The presence of opportunitic pathogens, including several Staphylococcus species and Propionibacterium acnes, was detected in both bioaerosol communities based on phylogenetic analysis. The isolation and identification of antibiotic-resistant bacteria from air samples also showed similar results with dominance of Actinobacteria and Proteobacteria in both hog farm and spray field air. Thus, the existence of opportunistic pathogens and antibiotic resistant bacteria in airborne communities evidences potential health risks to farmers and other residents from swine bioaerosol exposure.

• 한국토양비료학회지
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• 제44권1호
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• pp.127-145
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• 2011

Various environmental ecosystems are valuable sources for microbial ecology studies, and their analyses using recently developed molecular ecological approaches have drawn significant attention within the scientific community. Changes in the microbial community structures due to various anthropogenic activities can be evaluated by various culture-independent methods e.g. ARISA, DGGE, SSCP, T-RFLP, clone library, pyrosequencing, etc. Direct amplification of total community DNA and amplification of most conserved region (16S rRNA) are common initial steps, followed by either fingerprinting or sequencing analysis. Fingerprinting methods are relatively quicker than sequencing analysis in evaluating the changes in the microbial community. Being an efficient, sensitive and time- and cost effective method, T-RFLP is regularly used by many researchers to access the microbial diversity. Among various fingerprinting methods T-RFLP became an important tool in studying the microbial community structure because of its sensitivity and reproducibility. In this present review, we will discuss the important developments in T-RFLP methodology to distinguish the total microbial diversity and community composition in the various ecosystems.

• 미생물학회지
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• 제53권1호
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• pp.29-38
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• 2017

본 연구에서는 토판염전 결정지에 서식하는 세균군집의 계통학적 다양성을 분석하고 culturomics법에 기반하여 고도 호염균의 다양성을 확보하고자 하였다. 토판염전 내 세균밀도를 조사한 결과, 직접검경법에 의한 생균수는 평판배양법에 비해 $10^3{\sim}10^4$ 배 이상 높은 계수치를 나타내어 배양이 곤란한 세균(viable but non-culturable bacteria, VBNC)이 다수 존재해 있음으로 판단되었다. 토판염전 결정지 내 세균군집 다양성 해석을 위해 배양비의존적 방법인 pyrosequencing 분자기법을 이용하였다. 세균군집의 경우 1,778 OTUs, 다양성 지수 6.16로 나타났으며, 18문46강85목140과 243속으로 확인되었다. Archaea군집은 643 OTUs, 다양성 지수 4.95로 3문6강7목7과 38속이 분포해 있음이 확인되었다. 고도 호염균 생육에 적합한 배양배지 및 배양조건을 고려한 총 59가지의 다양한 배양 방법을 이용하여 137균주를 순수 분리하였다. 분리된 고도호염균의 16S rRNA 유전자 분석결과, 총4문11속의 다양한 계통군으로 확인되었으며 호염성 archaea 계통군 Haloterrigena 속과 haloferax 속이 culturomics법을 통해 성공적으로 분리되었다. 고도 호염균 다양성 확보를 위해 culturomics법이 매우 효과적임을 밝혔다.

• 농약과학회지
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• 제21권1호
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• pp.1-8
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• 2017

유기염소계 살충제 엔도설판은 2012년 국제적으로 사용이 금지되었으나 인도와 중국을 비롯한 국가들에서 여전히 사용되고 있으며 높은 잔류성과 이동성으로 인해 다양한 환경에서 검출되고 있다. 본 연구에서는 엔도설판이 토양세균의 군집구조에 미치는 영향을 16S rRNA 유전자 파이로시퀀싱 기법을 이용하여 분석하였다. 엔도설판을 100 mg/kg의 수준으로 밭토양에 처리했을 때 세균의 Operational taxonomic unit (OTU) 수와 다양성 지수가 감소했다가 서서히 증가하였으며 Proteobacteria와 Verrucomicrobia 문의 점유율은 증가하고 Chloroflexi와 Spirochaetes 문의 점유율은 감소한 것으로 나타나 엔도설판 처리가 토양세균 군집구조를 변화시키는 것을 확인하였다. 엔도설판 처리 시 Sphingomonas와 Burkholderia 속에 속하는 OTU의 점유율이 배양 초기에 높게 나타났으며 Pseudonocardia와 Opitutus 속에 속하는 OTU의 점유율은 배양 후기에 높게 나타났다. 이전 연구에서 Sphingomonas, Burkholderia, Pseudonocardia 속에 속하는 균주들이 엔도설판을 분해하였으므로 이 속에 속하는 OTU들이 토양에서 엔도설판의 분해에 단계적으로 관여했을 것으로 추정된다.