The influence of chlorine on marine bacterial communities was examined in this study. A non-chlorine-adapted marine bacterial community (NCAM) and a chlorine-adapted bacterial community (CAM, bacterial community treated with $0.2mg-Cl_2/L$ chlorine) were cultivated for 1 month. A distinct difference was observed between the NCAM and CAM, which shared only eight operational taxonomic units (OTUs), corresponding to 13.1% of the total number of identified OTUs. This result suggested that chlorine was responsible for the changes in the marine bacterial communities. Kordiimonas aquimaris was found to be a chlorine-resistant marine bacterium. The effect of intermittent chlorination on the two marine biofilm communities formed on the reverse osmosis (RO) membrane surface was investigated using various chlorine concentrations (0, 0.2, 0.4, 0.6 and 0.8 mg $Cl_2/L$). Although the average number of adherent marine bacteria on the RO membrane over a period of 7 weeks decreased with increasing chlorine concentration, disinfection efficiencies showed substantial fluctuations throughout the experiment. This is due to chlorine depletion that occurs during intermittent chlorination. These results suggest that intermittent chlorination is not an effective disinfection strategy to control biofilm formation.
RHO SANG CHUL;AN NAN HEE;AHN DAE HEE;LEE KYU HO;LEE DONG HUN;JAHNG DEOK JIN
Journal of Microbiology and Biotechnology
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v.15
no.2
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pp.287-295
/
2005
In order to compare bacteria] community structure and diversity in activated sludges, terminal restriction fragment length polymorphism (T-RFLP) of PCR-amplified 16s rDNAs was analyzed for 31 domestic and industrial wastewater treatment plants (WTPs). Regardless of the characteristics of the wastewaters, the bacteria] community structures of activated sludges appeared diverse and complex. In particular, activated sludges in domestic WTPs contained higher bacterial diversity than those in industrial WTPs. It was also found that terminal restriction fragment (T-RF) profiles derived from domestic WTPs were very similar with each other, although activated sludges were collected from different plants at different locations. Interestingly, activated sludges of a WTP where restaurant and toilet sewages of a company were managed showed a bacterial community structure similar to that of domestic WTPs. Activated sludges in leather industria] WTPs also showed a high similarity. However, other wastewaters possessed different bacterial communities, so that overall similarity was as low as about $30\%$. Since activated sludges from WTPs for domestic wastewaters and a company sewage appeared to hold similar bacterial communities, it was necessary to confirm if similar wastewaters induce a similar bacterial community. To answer this question, analysis of T-RFs for activated sludges, taken from another 12 domestic WTPs, was conducted by using a 6FAM$^{TM}$-Iabeled primer and an automated DNA sequencer for higher sensitivity. Among 12 samples, it was again found that T-RF profiles of activated sludges from Yongin, Sungnam, Suwon, and Tancheon domestic WTPs in Kyonggi-do were very similar with each other. On the other hand, T-RF profiles of activated sludges from Shihwa and Ansan WTPs were quite different from each other. It was thought that this deviation was caused by wastewaters, since Ansan and Shihwa WTPs receive both domestic and industrial wastewaters. From these results, it was tentatively concluded that similar bacterial communities might be developed in activated sludges, if WTPs treat similar wastewaters.
We have employed comparative RFL,P(Restriction Fragment Ixngth Pol~iniorphism) analysis and molecular phylogenetic techniques to investigate the diversity of uncultured microorganisms associated with the anaerobic PCP degradation in PCP-adapted enrichment cultures inoculated by samples from anaerobic cewage sludgc(Jangrim, Pusan) and leachate of landfill site(Kimhae). 16s rDNA cloncs were obtairted by PCR amplification of mixed population DNAs extracted directly from the nonactive and active stage ol each PCP-adapted culture. After three rounds of comparative RFLP analyses. two RFLP types. designated as Ala and Hld, were found prevalent and common in both active stage samples. Thc analysis of phylogenctic diversity bawd on the 5'-terminal 180 nt of sequences from whole clones of the Ala and Bld RFLP types showed close similarity among themselves. In case of Bld clones, 7XQ of them shared identical sequences. Thcse resuliq suggest that the clones of both RFLP types wcre originated from highly affiliated microorganisms which are e~iriched as a result of metabolic activity to PCP. The full-length 16s rRNA sequence of each representative clone from both RFLP types was determined. and an Ala clone w i n found to he related to Clo.strrdiurn ulfutzac~(Genk~ank No. Z69203) and a Bld clone to Thermobacteroides proteolyticus(Genbank No. X09335), with sequence similarities of 89%' and 97%. respectively.
Kim, Yeong-Kwan;Park, Sung-Gu;Lee, Dong-Hun;Choi, Sung-Chan
Journal of Korean Society of Water and Wastewater
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v.26
no.6
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pp.767-777
/
2012
Annular Biofilm Reactor (ABR) equipped with coupons of three different pipe materials (STS 304, PVC, PE) was used to generate drinking water biofilm samples. The level of assimilable organic carbon (AOC) during the sample generation period was $37.3{\mu}g/L$, and this level did not seem to be low enough to limit the formation of biofilm in this study. Terminal-restriction fragment length polymorphism (T-RFLP) analyses determined T-RF profile as early as 3 h of exposure on PVC coupons. Average surface roughness ($R_a$) measured by atomic force microscopic analyses was 125.7 nm for PVC, and this value was higher than for STS (71.6 nm) and PE (74.0 nm). However, biofilm formation was faster on STS (6 h) than on PE (12 h), which indicated that surface roughness might not be the only factor that controlled the initiation of biofilm development. Upon detection of the T-RF peaks, richness (S) and diversity indices such as Shannon (H) and Simpson (1/D) demonstrated a rather slow increase until 48 h followed by rapid increase regardless of the pipe materials. Differences of microbial community structures among the biofilm samples were determined based on the cluster analysis using Jaccard coefficients (Sj). Biofilm communities could be divided into two distinct groups according to the exposure time regardless of the pipe materials. First group contained a young (< 48 h) biofilm samples (10 out of 11) but second group contained a mature (${\geq}$ 48 h) samples (11 out of 14). Results suggested that, due to the complexity of biofilm, the targeting of the first group of cluster was crucial for optimizing the management of drinking water distribution systems and controlling microbial growth.
Effect of polycyclic aromatic hydrocarbons (PAHs) on the community structure of indigenous microorganisms in Gwangyang Bay sediments was investigated in Mar. & Aug.,2000. Microbial community structure was analyzed using 5'-terminal restriction fragment length polymorphism (T-RFLP) method. Microbial community structure based on T-RFLP method revealed that community differentiated by sampling period except station 1 located near the stream discharge site from Yeosu Industrial Complex. Even, microbial diversity was higher at stations showed relatively high concentrations of PAHs. The microbial community structure was severely changed during the enrichment culture with 1,000 ppm of PAHs mixture. It was also different between cultivated at 8$^{\circ}C$ and 30$^{\circ}C$. The results implied that temperature, poyosity, organic content and etc were more responsible than PAHs on the microbial community structure.
Ammonia-oxidizing bacteria (AOB) were enriched by repeating fed-batch cultivations in an AOB-selective medium of activated sludges from a domestic wastewater treatment plant. Enriched culture showed strong capabilities of ammonia oxidation [0.810 mg $NH_4^+$-N/mg mixed liquor suspended solids (MLSS)$\cdot$day] as well as $NO_x^-$-N production (0.617 mg $NO_x^-$-N/ mg MLSS$\cdot$day). Degree of enrichment was examined through fluorescent in situ hybridization (FISH) analyses using an AOB-specific Cy3-labeled oligonucleotide probe (NSOl90) and terminal-restriction fragment length polymorphism (T-RFLP) analyses. FISH analyses confirmed that the fraction of AOB among 4',6-diamidino-2-phenylindole (DAPI)-stained cells increased from about less than $0.001\%$ to approximately $42\%$ after enrichment of AOB, and T-RFLP analyses showed that bacterial community became simpler as enrichment was continued. When the enriched culture of AOB was added (150 mg/l as dry suspended solid) to the normal activated sludge (3,000 mg/l as dry suspended solid), nitrification efficiencies were improved from 0.020 mg $NO_x^-$-N/mg MLSS$\cdot$day to 0.041 mg $NO_x^-$-N/mg MLSS$\cdot$day in a synthetic wastewater and also from 0.0007 mg $NO_x^-$-N/mg MLSS$\cdot$day to 0.0918 mg $NO_x^-$-N/mg MLSS$\cdot$day in a real domestic wastewater. Therefore, it is expected that this enrichment method could be used for improving efficiency of nitrification in wastewater treatment plants.
This work aims to utilize wastes from the potato starch industry to produce single-cell protein (SCP) with high lysine content as animal feed. In this work, S-(2-aminoethyl)-L-cysteine hydrochloride-resistant Bacillus pumilus E1 was used to produce SCP with high lysine content, whereas Aspergillus niger was used to degrade cellulose biomass and Candida utilis was used to improve the smell and palatability of the feed. An orthogonal design was used to optimize the process of fermentation for maximal lysine content. The optimum fermentation conditions were as follows: temperature of 40℃, substrate concentration of 3%, and natural pH of about 7.0. For unsterilized potato starch wastes, the microbial communities in the fermentation process were determined by terminal restriction fragment length polymorphism analysis of bacterial 16S rRNA genes. Results showed that the dominant population was Bacillus sp. The protein quality as well as the amino acid profile of the final product was found to be significantly higher compared with the untreated waste product at day 0. Additionally, acute toxicity test showed that the SCP product was non-toxic, indicating that it can be used for commercial processing.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
/
pp.342-342
/
2017
Rice seeds are a home to endophytic bacterial communities which serve as a source of the plant's endophytes. As rice undergo physiological and adaptive modifications through cross breeding in the process of attaining salinity tolerance, this may also lead to changes in the endophytic bacterial community especially those residing in the seeds. This study explores the community structure of seed bacterial endophytes as influenced by rice parental lineage, genotype and physiological adaptation to salinity stress. Endophytic bacterial diversity was studied through culture dependent technique, cloning and Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Results revealed considerably diverse communities of bacterial endophytes in the interior of rice seeds. The richness of ribotypes ranges from 5-14 T-RFs corresponding to major groups of bacterial endophytes in the seeds. Endophytic bacterial diversity of the salt-sensitive IR29 is significantly more diverse compared to those of salt-tolerant cultivars. Proteobacteria followed by Actinobacteria and Firmicutes dominated the overall endophytic bacterial communities of the indica rice seeds based on 16S rDNA analysis of clones and isolates. Community profiles show common ribotypes found in all cultivars of the indica subspecies representing potential core microbiota belonging to Curtobacterium, Flavobacterium, Enterobacter, Xanthomonas, Herbaspirillum, Microbacterium and Stenotrophomonas. Multivariate analysis showed that the bacterial endophytic community and diversity of rice seeds are mainly influenced by their host's genotype, physiological adaptation to salt stress and parental lineage.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.244-244
/
2017
Soil salinity due to accumulation of salts particularly sodium chloride affects agricultural lands and their vegetation. Generally, rice is a moderately sensitive plant with some cultivars with varying tolerance to salinity. Though there are physiological differences between salt-sensitive and salt-tolerant rice cultivars, both are still affected especially during high salinity and prolonged exposure. This also ultimately affects their indigenous bacterial endophytes particularly those that inhabit the rice seed endosphere. This study investigates the dynamic structure of seed bacterial endophytes of salt-sensitive and tolerant rice cultivars grown in different levels of soil salinity. Endophytic bacterial diversity was studied Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis. Results revealed a very interesting pattern of diversity and shifts in community structure of bacterial endophytes in the rice seeds. There is a general decrease in diversity for the salt-sensitive rice cultivar, IR29 as soil salinity increases. For the salt-tolerant cultivars, IC32 and IC37, diversity interestingly increased at moderate salinity then decreased at high soil salinity. The patterns of community structure is also strikingly different for the salt-sensitive and salt-tolerant rice cultivars. IR29 has a more even distribution of abundance, but under soil salinity, the community shifted where Curtobacterium, Pantoea, Flavobacterium and Microbacterium become the more dominant bacterial communities. For IC32 and IC37, the dominant bacterial groups under normal stress conditions were also the dominant bacterial groups during salt stress conditions. Their seed bacterial community is dominated by endophytes belonging to Microbacterium, Flavobacterium, Pantoea, Kosakonia and Enterobacter. Stenotrophomonas and Xanthomonas have not changed in terms of abundance under different salinity stress level in the salt-sensitive and salt-tolerant rice cultivars. This study showed that soil salinity greatly influenced the seed bacterial communities of rice seeds irrespective of their physiological tolerance to salinity.
Kong, Hyun Gi;Kim, Nam Hee;Lee, Seung Yeup;Lee, Seon-Woo
The Plant Pathology Journal
/
v.32
no.2
/
pp.136-144
/
2016
Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.
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