For elucidating excessive growth of biofilm that subsequently leads to the clogging problem in a small town's sewer lines of Wisconsin, the FISH method was employed. At the beginning of the simulated experiments, ${\beta}$-subclass proteobacteria prevailed in runs fed with industrial wastewater, while ${\gamma}$-subclass proteobacteria dominated in runs with domestic wastewater. However, the bacterial community structure changed significantly over six weeks; Cytophaga-Flavobacterium (CF)group bacteria dominated in most runs fed with the small town's wastewater regardless of their source, while CF-group decreased strongly in run fed with domestic sewage from another city (Madison). It was also microscopically confirmed that most of those clogging materials was toilet tissue, which in turn may lead to vigorous growth of cellulose-degrading CF-group bacteria. This dominant presence of CF-group bacteria in the small town's sewer indicates that the main constituent of biofilm, toilet tissue (cellulose) in sewage, might have induced the unique pattern of their microbial community structure. Therefore, it suggests that molecular technique is useful for monitoring the clogging problems in sewer lines.
Nam, Ki Jung;Kim, Hyo-Jeong;Nam, Kyong-Hee;Pack, In Soon;Kim, Soo Young;Kim, Chang-Gi
Journal of Ecology and Environment
/
v.41
no.3
/
pp.99-106
/
2017
Background: Plants over-expressing Arabidopsis ABF3 (abscisic acid-responsive element-binding factor 3) have enhanced tolerance to various environmental stresses, especially drought. Using terminal restriction fragment length polymorphism (T-RFLP) analysis, we compared the rhizosphere-associated structures of microbial communities for transgenic potato containing this gene and conventional "Jopoong" plants. Results: During a 2-year field experiment, fungal richness, evenness, and diversity varied by year, increasing in 2010 when a moderate water deficit occurred. By contrast, the bacterial richness decreased in 2010 while evenness and diversity were similar in both years. No significant difference was observed in any indices for either sampling time or plant line. Although the composition of the microbial communities (defined as T-RF profiles) changed according to year and sampling time, differences were not significant between the transgenic and control plants. Conclusions: The results in this study suggest that the insertion of ABF3 into potato has no detectable (by current T-RFLP technique) effects on rhizosphere communities, and that any possible influences, if any, can be masked by seasonal or yearly variations.
Using a rotating biological contactor modified with a sequencing bath reactor system (SBRBC) designed and operated to remove phosphate and nitrogen [58], the microbial community structure of the biofilm from the SBRBC system was characterized based on the extracellular polymeric substance (EPS) constituents, electron microscopy, and molecular techniques. Protein and carbohydrate were identified as the major EPS constituents at three different biofilm thicknesses, where the amount of EPS and bacterial cell number were highest in the initial thickness of 0-100${\mu}m$. However, the percent of carbohydrate in the total amount of EPS decreased by about 11.23%, whereas the percent of protein increased by about 11.15% as the biofilm grew. Thus, an abundant quantity of EPS and cell mass, as well as a specific quality of EPS were apparently needed to attach to the substratum in the first step of the biofilm growth. A FISH analysis revealed that the dominant phylogenetic group was $\beta$- and $\gamma$-Proteobacteria, where a significant subclass of Proteobacteria for removing phosphate and/or nitrate was found within a biofilm thickness of 0-250${\mu}m$. In addition, 16S rDNA clone libraries revealed that Klebsiella sp. and Citrobacter sp. were most dominant within the initial biofilm thickness of 0-250${\mu}m$, whereas sulfur-oxidizing bacteria, such as Beggiatoa sp. and Thiothrix sp., were detected in a biofilm thickness over 250${\mu}m$. The results of the bacterial community structure analysis using molecular techniques agreed with the results of the morphological structure based on scanning electron microscopy. Therefore, the overall results indicated that coliform bacteria participated in the nitrate and phosphorus removal when using the SBRBC system. Moreover, the structure of the biofilm was also found to be related to the EPS constituents, as well as the nitrogen and phosphate removal efficiency. Consequently, since this is the first identification of the bacterial community and structure of the biofilm from an RBC simultaneously removing nitrogen and phosphate from domestic wastewater, and it is hoped that the present results may provide a foundation for understanding nitrate and phosphate removal by an RBC system.
Oil spill was found in 1999 from a diesel storage facility located near the top of Baekun Mountain in Uiwang City. Application of bioremediation techniques was very relevant in removing oil spills in this site, because the geological condition was not amenable for other onsite remediation techniques. For efficient bioremediation, bacterial communities of the contaminated site and the uncontaminated control site were compared using both molecular and cultivation techniques. Soil bacterial populations were observed to be stimulated to grow in the soils contaminated with diesel hydrocarbon, whereas fungal and actinomycetes populations were decreased by diesel contamination. Most of the dieseldegrading bacteria isolated from contaminated forest soils were strains of Pseudomonas, Ralstonia, and Rhodococcus species. Denaturing gradient gel electrophoresis (DGGE) analysis revealed that the profiles were different among the three contaminated sites, whereas those of the control sites were identical to each other. Analysis of 16S rDNA sequences of dominant isolates and clones showed that the bacterial community was less diverse in the oil-contaminated site than at the control site. Sequence analysis of the alkane hydroxylase genes cloned from soil microbial DNAs indicated that their diversity and distribution were different between the contaminated site and the control site. The results indicated that diesel contamination exerted a strong selection on the indigenous microbial community in the contaminated site, leading to predominance of well-adapted microorganisms in concurrence with decrease of microbial diversity.
Global warming has accelerated glacial retreat in the high Arctic. The exposed glacier foreland is an ideal place to study chronosequential changes in ecosystems. Although vegetation succession in the glacier forelands has been studied intensively, little is known about the microbial community structure in these environments. Therefore, this study focused on how glacial retreat influences the bacterial community structure and its relationship with soil properties. This study was conducted in the foreland of the Midtre Lovénbreen glacier in Svalbard (78.9°N). Seven soil samples of different ages were collected and analyzed for moisture content, pH, soil organic carbon and total nitrogen contents, and soil organic matter fractionation. In addition, the structure of the bacterial community was determined via pyrosequencing analysis of 16S rRNA genes. The physical and chemical properties of soil varied significantly along the distance from the glacier; with increasing distance, more amounts of clay and soil organic carbon contents were observed. In addition, Cyanobacteria, Firmicutes, and Actinobacteria were dominant in soil samples taken close to the glacier, whereas Acidobacteria were abundant further away from the glacier. Diversity indices indicated that the bacterial community changed from homogeneous to heterogeneous structure along the glacier chronosequence/distance from the glacier. Although the bacterial community structure differed on basis of the presence or absence of plants, the soil properties varied depending on soil age. These findings suggest that bacterial succession occurs over time in glacier forelands but on a timescale that is different from that of soil development.
Kimchi is a traditional Korean fermented vegetable food, the production of which involves brining of Korean cabbage, blending with various other ingredients (red pepper powder, garlic, ginger, salt-pickled seafood, etc.), and fermentation. Recently, kimchi has also become popular in the Western world because of its unique taste and beneficial properties such as antioxidant and antimutagenic activities, which are derived from the various raw materials and secondary metabolites of the fermentative microorganisms used during production. Despite these useful activities, analysis of the microbial community present in kimchi has received relatively little attention. The objective of this study was to evaluate the bacterial community structure from the raw materials, additives, and final kimchi product using the culture-independent method. Specifically, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to analyze the 16S rRNA partial sequences of the microflora. One primer set for bacteria, 341FGC-518R, reliably produced amplicons from kimchi and its raw materials, and these bands were clearly separated on a 35-65% denaturing gradient gel. Overall, 117 16S rRNA fragments were identified by PCR-DGGE analysis. Pediococcus pentosaceus, Leuconostoc citreum, Leuconostoc gelidum, and Leuconostoc mesenteroides were the dominant bacteria in kimchi. The other strains identified were Tetragenococcus, Pseudomonas, Weissella, and uncultured bacterium. Comprehensive analysis of these microorganisms could provide a more detailed understanding of the biologically active components of kimchi and help improve its quality. PCR-DGGE analysis can be successfully applied to a fermented food to detect unculturable or other species.
Ranhee Lee;Gwangsu Ha;Ho Jin Jeong;Do-Youn Jeong;Hee-Jong Yang
Journal of Life Science
/
v.34
no.4
/
pp.254-263
/
2024
Korean jang is a food made using fermented soybeans, and the typical products include gochujang (GO), doenjang (DO), cheonggukjang (CH), and ganjang (GA). In this study, 16S rRNA metagenome analysis was performed on a total of 200 types of GO, DO, CH, and GA using next-generation sequencing to analyze the microbial community of fermented soybean foods and compare taxonomic (biomarker) differences. Alpha diversity analysis showed that in the CHAO index, the species richness index tended to be significantly higher compared to the DO and GA groups (p<0.001). The results of the microbial distribution analysis of the GO, DO, CH, and GA products showed that at the order level, Bacillales was the most abundant in the GO, DO, and CH groups, but Lactobacillales was most abundant in the GA group. Linear discriminant analysis effect (LEfSe) analysis was used to identify biomarkers at the family and species levels. Leuconostocaceae, Thermoactinomycetaceae, Bacillaceae, and Enterococcaceae appeared as biomarkers at the family level, and Bacillus subtilis, Kroppenstedtia sanguinis, Bacillus licheniformis, and Tetragenococcus halophilus appeared at the species level. Permutational multivariate analysis of variance (PERMANOVA) analysis showed that there was a significant difference in the microbial community structure of the GO, DO, CH, and GA groups (p=0.001), and the microbial community structure of the GA group showed the greatest difference. This study clarified the correlation between the characteristics of Korean fermented foods and microbial community distribution, enhancing knowledge of microorganisms participating in the fermentation process. These results could be leveraged to improve the quality of fermented soybean foods.
T-RFLP analysis and clone sequencing analysis based on bacterial 16S rDNA were conducted to assess bacterial community structure and diversity in Zoysia japonica soil treated with liquid fertilizer containing amino acids(LFcAA) after spray with herbicide. The results of T-RFLP (terminal restriction fragment length poly-morphism) analysis using restriction enzyme Hae III showed that the T-RFs of various size appeared evenly in the 32 clones of KD3 and 38 clones of KD4 respectively that had been treated with liquid fertilizer containing amino acid(LFcAA) compared to 23 clones of KD2 hat had not been treated with LFcAA. The microbial com- munity structure in KD2 appeared less diverse than those in KD3 and KD4. Analysis of partial sequences for 110 clones from KDI (control), KD2 (non-treated), KD3 (LFcAA 1X), KD4 (LFcAA 2X), respectively, revealed that most bacteria were related with uncultured bacteria in a 16S rDNA sequence similarity range of 91-99% through blast search. Otherwise, the other clones were members of proteobacteria, Acidobacteria, Act-inobacteria, Sphingobacteria and Planctomyces groups. Especially in KD4, members of Alpha Proteobacteria, Rhizobiales, Sphigomonadales, Caulobacterales, Gamma Proteobacteria, the genus Pseudomonas, Betapro-teobacteria, Nitrosomonadales and genus Nitrosospira appeared to be dominant. In addition, Acidobacteria group, Actinobacteria group, Planctomycetacia and Sphingobacteria were also shown. The microbial com-munity structure in Z. japonica soil sprayed with herbicide was affected by LFcAA.
Although many studies on the effects of genetically modified (GM) crops on soil microorganisms have been carried out over the past decades, they have provided contradictory information, even for the same GM crop, owing to the diversity of the soil environments in which they were conducted. This inconsistency in results suggests that the effects of GM crops on soil microorganisms should be considered from many aspects. In this study, we investigated the effects of the GM drought-tolerant rice MSRB2-Bar-8, which expresses the CaMSRB2 gene, on soil microorganisms based on the culture-dependent and culture-independent methods. To this end, rhizosphere soils of GM and non-GM (IM) rice were analyzed for soil chemistry, population densities of soil microorganisms, and microbial community structure (using pyrosequencing technology) at three growth stages (seedling, tillering, and maturity). There was no significant difference in the soil chemistry between GM and non-GM rice. The microbial densities of the GM soils were found to be within the range of those of the non-GM rice. In the pyrosequencing analyses, Proteobacteria and Chloroflexi were dominant at the seedling stage, while Chloroflexi showed dominance over Proteobacteria at the maturity stage in both the GM and non-GM soils. An UPGMA dendrogram showed that the soil microbial communities were clustered by growth stage. Taken together, the results from this study suggest that the effects of MSRB2-Bar-8 cultivation on soil microorganisms are not significant.
This study was performed to evaluate effect of different fertilization management practices on soil microbial activities and community structure using soil enzyme activities and PLFA contents in volcanic ash citrus orchard soil. Six experiment plots have differently managed based on the recommended application rate(NPK) of chemical fertilizer and compost for 13 years. Experiment plots were composed of no-fertilization(control), compost only, half amount of NPK with compost (1/2NPK+COM), NPK, NPK with compost(NPK+COM), and 3 times amount of NPK(3NPK). Soil samples collected in early March, May, July, and September 2007. Urease activity was high at NPK+COM in March, May, and September. It was higher in NPK+COM than in NPK. Urease activity decreased according to the order NPK>compost>control in March and May; compost>NPK>control in July and September. Dehydrogenase activity was significantly higher in 1/2NPK+COM($4.3ug\;TPF\;g^{-1}\;24h^{-1}$) than in control($2.4ug\;TPF\;g^{-1}\;24h^{-1}$), May. $\beta$-glucosidase activity was significantly higher in NPK and 1/2NPK+COM than in control, May. In March, Total PLFA contents were higher in NPK+COM($349.2n\;mol\;g^{-1}$) than in 3NPK($228.5n\;mol\;g^{-1}$). And that were higher in 1/2NPK+COM($237.8n\;mol\;g^{-1}$) than in 3NPK($133.1n\;mol\;g^{-1}$), May. Distribution ratio of soil microbial groups by PLFA biomaker were not significantly difference in between seasonal and treatments. Principal component analysis by PLFA profiles showed that microbial community in compost and 3NPK plot were different compared with other treatments in March. But Differences in compost and 3NPK plot were not found in May. Our result showed that the change of microbial community structure affected by fertilization effect and seasonable variation.
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