• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.118-118
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• 2008

The phylogenetic diversity of microbial communities in calcareous mats from Spearfish Creek, a freshwater stream located in the Black Hills of South Dakota, was examined using PCR-based 16S rDNA sequence analysis. In this study, two types of calcareous mats were compared: mature mats formed on the natural substrate of rock surfaces and developing mats on an artificial substrate of glass slides. Among 63 unique isolates from a clone library of 16S rRNA genes from mature mat samples, there were 8 phyla of Bacteria represented. The predominant phylum was Proteobacteria (48%), with the $\beta$ subclass being the largest group. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes from slide samples collected at intervals for four months showed considerable diversity of the microbial community from the earliest stages of community development. Amplicons isolated from DGGE gels and sequenced indicated that community succession has occurred without increasing microbial diversity. However, light microscopic analysis revealed a significant increase in microbial cell density throughout the community development. Scanning electron microscopy of mat samples provides evidence that diatoms are also important members of calcareous mat communities.

• Journal of Microbiology
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• v.44 no.2
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• pp.155-161
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• 2006

Comparative analysis of microbial communities in a sequencing batch reactor which performed enhanced biological phosphorus removal (EBPR) was carried out using a cultivation-based technique and 16S rRNA gene clone libraries. A standard PCR protocol and a modified PCR protocol with low PCR cycle was applied to the two clone libraries of the 16S rRNA gene sequences obtained from EBPR sludge, respectively, and the resulting 424 clones were analyzed using restriction fragment length polymorphisms (RFLPs) on 16S rRNA gene inserts. Comparison of two clone libraries showed that the modified PCR protocol decreased the incidence of distinct fragment patterns from about 63 % (137 of 217) in the standard PCR method to about 34 % (70 of 207) under the modified protocol, suggesting that just a low level of PCR cycling (5 cycles after 15 cycles) can significantly reduce the formation of chimeric DNA in the final PCR products. Phylogenetic analysis of 81 groups with distinct RFLP patterns that were obtained using the modified PCR method revealed that the clones were affiliated with at least 11 phyla or classes of the domain Bacteria. However, the analyses of 327 colonies, which were grouped into just 41 distinct types by RFLP analysis, showed that they could be classified into five major bacterial lineages: ${\alpha},\;{\beta},\;{\gamma}-$ Proteobacteria, Actinobacteria, and the phylum Bacteroidetes, which indicated that the microbial community yielded from the cultivation-based method was still much simpler than that yielded from the PCR-based molecular method. In this study, the discrepancy observed between the communities obtained from PCR-based and cultivation-based methods seems to result from low culturabilities of bacteria or PCR bias even though modified culture and PCR methods were used. Therefore, continuous development of PCR protocol and cultivation techniques is needed to reduce this discrepancy.

• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.645-658
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• 2021

Porins are essential for the viability of Gram-negative bacteria. They ensure the uptake of nutrients, can be involved in the maintenance of outer membrane integrity and define the antibiotic or drug resistance of organisms. The function and structure of porins in proteobacteria is well described, while their function in photoautotrophic cyanobacteria has not been systematically explored. We compared the domain architecture of nine putative porins in the filamentous cyanobacterium Anabaena sp. PCC 7120 and analyzed the seven candidates with predicted OprB-domain. Single recombinant mutants of the seven genes were created and their growth capacity under different conditions was analyzed. Most of the putative porins seem to be involved in the transport of salt and copper, as respective mutants were resistant to elevated concentrations of these substances. In turn, only the mutant of alr2231 was less sensitive to elevated zinc concentrations, while mutants of alr0834, alr4741 and all4499 were resistant to high manganese concentrations. Notably the mutant of alr4550 shows a high sensitivity against harmful compounds, which is indicative for a function related to the maintenance of outer membrane integrity. Moreover, the mutant of all5191 exhibited a phenotype which suggests either a higher nitrate demand or an inefficient nitrogen fixation. The dependency of porin membrane insertion on Omp85 proteins was tested exemplarily for Alr4550, and an enhanced aggregation of Alr4550 was observed in two omp85 mutants. The comparative analysis of porin mutants suggests that the proteins in parts perform distinct functions related to envelope integrity and solute uptake.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.26 no.2
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• pp.99-115
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• 2023

Purpose: Exclusive breastfeeding promotes gut microbial compositions associated with lower rates of metabolic and autoimmune diseases. Its cessation is implicated in increased microbiome-metabolome discordance, suggesting a vulnerability to dietary changes. Formula supplementation is common within our low-income, ethnic-minority community. We studied exclusively breastfed (EBF) neonates' early microbiome-metabolome coupling in efforts to build foundational knowledge needed to target this inequality. Methods: Maternal surveys and stool samples from seven EBF neonates at first transitional stool (0-24 hours), discharge (30-48 hours), and at first appointment (days 3-5) were collected. Survey included demographics, feeding method, medications, medical history and tobacco and alcohol use. Stool samples were processed for 16S rRNA gene sequencing and lipid analysis by gas chromatography-mass spectrometry. Alpha and beta diversity analyses and Procrustes randomization for associations were carried out. Results: Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria were the most abundant taxa. Variation in microbiome composition was greater between individuals than within (p=0.001). Palmitic, oleic, stearic, and linoleic acids were the most abundant lipids. Variation in lipid composition was greater between individuals than within (p=0.040). Multivariate composition of the metabolome, but not microbiome, correlated with time (p=0.030). Total lipids, saturated lipids, and unsaturated lipids concentrations increased over time (p=0.012, p=0.008, p=0.023). Alpha diversity did not correlate with time (p=0.403). Microbiome composition was not associated with each samples' metabolome (p=0.450). Conclusion: Neonate gut microbiomes were unique to each neonate; respective metabolome profiles demonstrated generalizable temporal developments. The overall variability suggests potential interplay between influences including maternal breastmilk composition, amount consumed and living environment.

• Fisheries and Aquatic Sciences
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• v.27 no.8
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• pp.525-532
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• 2024

The strategy of cultivating healthy fish by manipulating their gut microbiota composition using probiotics is crucial for establishing sustainable fish farms. Information on the gut microbiota composition of healthy fish is important for the efficient development of probiotics and the aquaculture strategies that would apply them. This study assessed the gut microbiota composition of farmed olive flounder according to the growth stage to provide useful information for the development of probiotics. The fish were classified according to their weight, and the intestines of five juveniles weighing less than 100 g and five subadults weighing between 400 and 800 g were used for analysis. The results of alpha diversity analysis confirmed higher richness and diversity in the subadult group than in the juvenile group. Beta diversity analysis revealed clear boundaries and distances between the groups; however, individuals within each group were similar. The two groups showed marked differences in their gut microbiota composition. Phylum-level analysis revealed that the most abundant phylum was Proteobacteria in the juvenile group and Firmicutes in the subadult group. Notably, genus-level analysis revealed that Vibrio accounted for more than 50% of the total composition in the juvenile group. Linear discriminant analysis effect size revealed that the genera Vibrio, Photobacterium, and Leuconostoc were representative of the juvenile group, whereas 19 genera, including Lactococcus and Vagococcus, were representative of the subadult group. Thus, information on the representative microorganisms abundantly present in the intestines of healthy olive flounder can be considered for the selection and development of a probiotic genus for olive flounder farming.

• Korean Journal of Microbiology
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• v.53 no.2
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• pp.83-96
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• 2017

A constructed sea stream in Yeongdo, Busan, Republic of Korea is mostly static due to the lifted stream bed and tidal characters, and receives domestic wastewater nearby, causing a consistent odor production and water quality degradation. Bioaugmentation of a microbial consortium was proposed as an effective and economical restoration technology to restore the polluted stream. The microbial consortium activated on site was augmented on a periodic basis (7~10 days) into the most polluted site (Site 2) which was chosen considering the pollution level and tidal movement. Physicochemical parameters of water qualities were monitored including pH, temperature, DO, ORP, SS, COD, T-N, and T-P. COD and microbial community analyses of the sediments were also performed. A significant reduction in SS, COD, T-N, and COD (sediment) at Site 2 occurred showing their removal rates 51%, 58% and 27% and 35%, respectively, in 13 months while T-P increased by 47%. In most of the test sites, population densities of sulfate reducing bacterial (SRB) groups (Desulfobacteraceae_uc_s, Desulfobacterales_uc_s, Desulfuromonadaceae_uc_s, Desulfuromonas_g1_uc, and Desulfobacter postgatei) and Anaerolinaeles was observed to generally decrease after the bioaugmentation while those of Gamma-proteobacteria (NOR5-6B_s and NOR5-6A_s), Bacteroidales_uc_s, and Flavobacteriales_uc_s appeared to generally increase. Aerobic microbial communities (Flavobacteriaceae_uc_s) were dominant in St. 4 that showed the highest level of DO and least level of COD. These microbial communities could be used as an indicator organism to monitor the restoration process. The alpha diversity indices (OTUs, Chao1, and Shannon) of microbial communities generally decreased after the augmentation. Fast uniFrac analysis of all the samples of different sites and dates showed that there was a similarity in the microbial community structures regardless of samples as the augmentation advanced in comparison with before- and early bioaugmentation event, indicating occurrence of changing of the indigenous microbial community structures. It was concluded that the bioaugmentation could improve the polluted water quality and simultaneously change the microbial community structures via their niche changes. This in situ remediation technology will contribute to an eco-friendly and economically cleaning up of polluted streams of brine water and freshwater.

• Journal of Microbiology
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• v.42 no.4
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• pp.285-291
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• 2004

The soil bacterial community and some inoculated bacteria were monitored to assess the microbial responses to prescribed fire in their microcosm. An acridine orange direct count of the bacteria in the unburned control soil were maintained at a relatively stable level $(2.0\~2.7\times10^9\;cells/g^{-1}{\cdot}soil)$ during the 180 day study period. The number of bacteria in the surface soil was decreased by fire, but was restored after 3 months. Inoculation of some bacteria increased the number of inoculated bacteria sev­eral times and these elevated levels lasted several months. The ratios of eubacteria detected by a flu­orescent in situ hybridization (FISH) method to direct bacterial count were in the range of $60\~80\%$ during the study period, with the exception of some lower values at the beginning, but there were no definite differences between the burned and unburned soils or the inoculated and uninoculated soils. In the unburned control soil, the ratios of $\alpha-,\beta-\;and\;\gamma-subgroups$ of the proteobacteria, Cytophaga-Fla­vobacterium and other eubacteria groups to that of the entire eubacteria were 13.7, 31.7, 17.1, 16.8 and $20.8\%,$ respectively, at time 0. The overall change on the patterns of the ratios of the 5 subgroups of eubacteria in the uninoculated burned and inoculated soils were similar to those of the unburned con­trol soil, with the exception of some minor variations during the initial period. The proportions of each group of eubacteria became similar in the different microcosms after 6 months, which may indicate the recovery of the original soil microbial community structure after fire or the inoculation of some bac­teria. The populations of Azotobacter vinelandii, Bacillus megaterium and Pseudomonas fluorescens, which had been inoculated to enhance the microbial activities, and monitored by FISH method, showed similar changes in the microcosms, and maintained high levels for several months.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.83-89
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• 2013

A bacterium, isolated from a vegetable field in a plastic film house and named strain CY6 was capable of biodegrading chlorpyrifos (CP). Based on the phenotypic features and the phylogenetic similarity of 16S rRNA gene sequences, strain CY6 was identified as a Naxibacter sp.. CP was utilized as the sole source of carbon and phosphorus by Naxibacter sp. CY6. We examined the role of this Naxibacter sp. in the degradation of other OP insecticides under liquid cultures. Parathion, methyl parathion, diazinon, cadusafos, and ethoprop could also be degraded by Naxibacter sp. CY6 when they are provided as the sole sources of carbon and phosphorus. Additionally, Naxibacter sp. CY6 ($10^8$ CFU/g) added to soil with CP (100 mg/kg) resulted in a higher degradation rate of approximately 90% than the rate obtained from uninoculated soils. These results highlight the potential of this bacterium to be used in the cleanup of contaminated pesticide soil.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1656-1664
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• 2009

Bioaugmentation of bioreactors focuses on the removal of numerous organics, with little attention typically paid to the maintenance of high and stable nitrite accumulation in partial nitrification. In this study, a bioaugmented membrane bioreactor (MBR) inoculated with enriched ammonia-oxidizing bacteria (AOB) was developed, and the effects of dissolved oxygen (DO) and temperature on the stability of partial nitrification and microbial community structure, in particular on the nitrifying community, were evaluated. The results showed that DO and temperature played the most important roles in the stability of partial nitrification in the bioaugmented MBR. The optimal operation conditions were found at 2-3 mgDO/l and $30^{\circ}C$, achieving 95% ammonia oxidization efficiency and nitrite ratio ($NO_2^-/{NO_x}^-$) of 0.95. High DO (5-6 mg/l) and low temperature ($20^{\circ}C$) had negative impacts on nitrite accumulation, leading to nitrite ratio drop to 0.6. However, the nitrite ratio achieved in the bioaugmented MBR was higher than that in most previous literatures. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) were used to provide an insight into the microbial community. It showed that Nitrosomonas-like species as the only detected AOB remained predominant in the bioaugmented MBR all the time, and coexisted with numerous heterotrophic bacteria. The heterotrophic bacteria responsible for mineralizing soluble microbial products (SMP) produced by nitrifiers belonged to the Cytophaga-Flavobacterium-Bacteroides (CFB) group, and $\alpha$-, $\beta$-, and $\gamma$- Proteobacteria. The fraction of AOB ranging from 77% to 54% was much higher than that of nitrite-oxidizing bacteria (0.4-0.9%), which might be the primary cause for the high and stable nitrite accumulation in the bioaugmented MBR.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.175-182
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• 2016

In this study, we isolated aromatic compounds (lignin polymers) utilizing bacteria in humus layer of oak forest and investigated phylogenetic characteristics and correlation with major bacterial populations in the humus layer by pyrosequencing. Forty-two isolates using aromatic compounds such as p-anisic acid, benzoic acid, ferulic acid and p-coumaric acid were isolated and phylogentic analyses based on 16S rRNA gene sequences showed that the isolates belonged to the genus Rhizobium, Sphingomonas, Burkhorlderia, and Pseudomonas. Among these, Burkhorlderia species which belong to Betaproteobacteria class occupied 83% among the isolates. The bacterial populations in humus layer of oak forest were characterized by next generation pyrosequencing based on 16S rRNA gene sequences. The humus sample produced 7,862 reads, 1,821 OTUs and 6.76 variability index with 97% of significance level, respectively. Bacterial populations consist of 22 phyla and Betaproteobacteria were the major phylum consisting of 15 genera including Burkholderia, Polaromonas, Ralstoria, Zoogloea, and Variovorax. Approximately fifty percentage of them was Burkholderia. Burkholderia as the majority of population in the humus was considered to play a role in degrading lignin in humus layer of oak forest.