• Title/Summary/Keyword: Anaerobic sequencing

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Biological Nitrogen and Phosphorus Removal Characteristics on Organic Material and Nitrate Loadings in SBR Process (연속회분식반응조에서 유기물 부하와 질산염농도에 따른 생물학적 질소 및 인 제거 특성)

  • Kim, I-Tae;Lee, Hee-Ja;Kim, Kwang-Soo;Bae, Woo-keun
    • Journal of Korean Society on Water Environment
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    • v.20 no.6
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    • pp.571-576
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    • 2004
  • Since anaerobic/anoxic/oxic process, which is a typical mainstream biological nitrogen and phosphorus removal process, utilizes influent organic matter as an external carbon source for phosphorus release in anaerobic or anoxic stage, influent COD/T-P ratio gives a strong influence on performance of phosphorus removal process. In this study, a bench scale experiment was carried out for SBR process to investigate nitrogen and phosphorus removal at various influent COD/T-P ratio and nitrate loadings of 23~73 and 1.6~14.3g $NO_3{^-}-N/kg$ MLSS, respectively. The phosphorus release and excess uptake in anoxic condition were very active at influent COD/T-P ratios of 44 and 73. However, its release and uptake was not obviously observed at COD/T-P ratio of 23. Consequently, phosphorus removal efficiency was decreased. In addition, the phosphorus release and uptake rate in anoxic condition increased as the nitrate loading decreased. Specific denitrification rate had significantly high correlation with organic materials and nitrate loadings of the anoxic phase too. The rate of phosphorus release and uptake in the anoxic condition were $0.08{\sim}0.94kg\;S-P/kg\;MLSS{\cdot}d$ and $0.012{\sim}0.1kg\;S-P/kg\;MLSS{\cdot}d$, respectively.

Successful Enrichment of Rarely Found Candidatus Anammoxoglobus propionicus from Leachate Sludge

  • Hsu, Shu-Chuan;Lai, Yen-Chun;Hsieh, Ping-Heng;Cheng, Pun-Jen;Wong, Suen-Shin;Hung, Chun-Hsiung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.879-887
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    • 2014
  • Bacteria that mediate the anaerobic oxidation of ammonium (anammox) have been detected in natural ecosystems, as well as various wastewater treatment systems. In this study, sludge from a particular landfill leachate anaerobic treatment system was selected as the incubation seed for anammox microorganism enrichment owing to its possible anammox activity. Transmission electron microscopy observation, denaturing gradient gel electrophoresis analysis, and cloning/sequencing techniques were applied to identify the diversity of anammox microorganisms throughout the incubation. During the early stage of operation, the diversity of anammox microorganisms was similar to the original complex microbes in the seed sludge. However, as incubation time increased, the anammox microorganism diversity within the system that was originally dominated by Candidatus (Ca.) Brocadia sp. was replaced by Ca. Anammoxoglobus propionicus. The domination of Ca. Anammoxoglobus propionicus produced a stable removal of ammonia (70 mg-N/l) and nitrite (90 mg-N/l), and the total nitrogen removal efficiency was maintained at nearly 95%. The fluorescence in situ hybridization results showed that Ca. Anammoxoglobus propionicus was successfully enriched from $1.8{\pm}0.6%$ initially to $65{\pm}5%$ after 481 days of operation. Therefore, the present results demonstrated the feasibility of enriching Ca. Anammoxoglobus propionicus from leachate sludge, even though the original cell count was extremely low. Application of this seldom found anammox organism could offer an alternative to current ammonia-nitrogen treatment.

Anaerobic Ammonium-Oxidizing Bacteria in Cow Manure Composting

  • Wang, Tingting;Cheng, Lijun;Zhang, Wenhao;Xu, Xiuhong;Meng, Qingxin;Sun, Xuewei;Liu, Huajing;Li, Hongtao;Sun, Yu
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1288-1299
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    • 2017
  • Composting is widely used to transform waste into valuable agricultural organic fertilizer. Anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the global nitrogen cycle, but their role in composting remains poorly understood. In the present study, the community structure, diversity, and abundance of anammox bacteria were analyzed using cloning and sequencing methods by targeting the 16S rRNA gene and the hydrazine oxidase gene (hzo) in samples isolated from compost produced from cow manure and rice straw. A total of 25 operational taxonomic units were classified based on 16S rRNA gene clone libraries, and 14 operational taxonomic units were classified based on hzo gene clone libraries. The phylogenetic tree analysis of the 16S rRNA gene and deduced HZO protein sequences from the corresponding encoding genes indicated that the majority of the obtained clones were related to the known anammox bacteria Candidatus "Brocadia," Candidatus "Kuenenia," and Candidatus "Scalindua." The abundances of anammox bacteria were determined by quantitative PCR, and between $2.13{\times}10^5$ and $1.15{\times}10^6$ 16S rRNA gene copies per gram of compost were found. This study provides the first demonstration of the existence of anammox bacteria with limited diversity in cow manure composting.

Changes in Gut Microbial Community of Pig Feces in Response to Different Dietary Animal Protein Media

  • Jeong, Yujeong;Park, Jongbin;Kim, Eun Bae
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1321-1334
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    • 2020
  • Beef, pork, chicken and milk are considered representative protein sources in the human diet. Since the digestion of protein is important, the role of intestinal microflora is also important. Despite this, the pure effects of meat and milk intake on the microbiome are yet to be fully elucidated. To evaluate the effect of beef, pork, chicken and milk on intestinal microflora, we observed changes in the microbiome in response to different types of dietary animal proteins in vitro. Feces were collected from five 6-week-old pigs. The suspensions were pooled and inoculated into four different media containing beef, pork, chicken, or skim milk powder in distilled water. Changes in microbial communities were analyzed using 16S rRNA sequencing. The feces alone had the highest microbial alpha diversity. Among the treatment groups, beef showed the highest microbial diversity, followed by pork, chicken, and milk. The three dominant phyla were Proteobacteria, Firmicutes, and Bacteroidetes in all the groups. The most abundant genera in beef, pork, and chicken were Rummeliibacillus, Clostridium, and Phascolarctobacterium, whereas milk was enriched with Streptococcus, Lactobacillus, and Enterococcus. Aerobic bacteria decreased while anaerobic and facultative anaerobic bacteria increased in protein-rich nutrients. Functional gene groups were found to be over-represented in protein-rich nutrients. Our results provide baseline information for understanding the roles of dietary animal proteins in reshaping the gut microbiome. Furthermore, growth-promotion by specific species/genus may be used as a cultivation tool for uncultured gut microorganisms.

Isolation and Characteristics of a Homofermentative lactic Acid Bacterium (호모발효 젖산군의 분리 및 특성)

  • 하미영;정선용;김성준
    • KSBB Journal
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    • v.17 no.4
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    • pp.333-338
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    • 2002
  • This study was targeted to isolate and characterize a bacterium producing lactic acid in a large amount. Lactic acid bacteria of about fifty strains were isolated from kimchi, a Korean traditional fermented vegetable food. Strain KH-1 of them was most effective in the lactic acid production and showed 99% homology with Lactobacillus casei from analysis of 16S rRNA sequencing. The conversion ratio of lactic acid from glucose by 1. casei KH-1 was 98% in anaerobic condition, and the lactic acid was composed as racemic mixture of D(-)-and L(+)-lactic acid, 7% and 93%, respectively. This result indicated that L. casei KH-1 was a homofermentative bacterium mainly producing L(+)-lactic acid. The strain KH-1 used glucose as a preferential substrate but not utilized lactose. In investigation of more inexpensive nitrogen source for cultivation of strain KH-1 using industrial MRS medium, when yeast extract and corn steep liquor were used at the ratio of 1 to 1, the molar yield of lactic acid produced per mole of glucose(Yp/s) was 1.09.

High-rate Denitrifying Process Based on Methanol and Characteristics of Organic Carbon Uptake (메탄올 기반 탈질 공정의 고속화 및 탄소 섭취 특성)

  • Park, Suin;Jeon, Junbeom;Bae, Hyokwan
    • Journal of Korean Society on Water Environment
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    • v.36 no.6
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    • pp.581-591
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    • 2020
  • In this study, two types of reactors were operated to examine the properties of methanol uptake under the high-rate denitrification process. In a sequencing batch reactor, the denitrifying activity was enriched up to 0.80 g-N/g-VSS-day for 72 days. Then, the enriched denitrifying sludge was transferred to a completely stirred tank reactor (CSTR). At the final phase on Day 46-50, the nitrogen removal efficiency was around 100% and the total nitrogen removal rate reached 0.097±0.003 kg-N/㎥-day. During the continuous process, the sludge settling index (SVI30) was stabilized as 118.3 mL/g with the biomass concentration of 1,607 mg/L. The continuous denitrifying process was accelerated by using a sequencing batch reactor (SBR) with a total nitrogen removal rate of 0.403±0.029 kg-N/㎥-day with a high biomass concentration of 8,433 mg-VSS/L. Because the reactor was open to ambient air with the dissolved oxygen range of 0.2-0.5 mg-O2/L, an increased organic carbon requirement of 5.58±0.70 COD/NO3--N was shown for the SBR in comparison to the value of 4.13±0.94 for the test of the same biomass in a completely anaerobic batch reactor. The molecular analysis based on the 16S rRNA gene showed that Methyloversatilis discipulorum and Hyphomicrobium zavarzinii were the responsible denitrifiers with the sole organic carbon source of methanol.

Decoding the intestinal microbiota repertoire of sow and weaned pigs using culturomic and metagenomic approaches

  • Mun, Daye;Kim, Hayoung;Shin, Minhye;Ryu, Sangdon;Song, Minho;Oh, Sangnam;Kim, Younghoon
    • Journal of Animal Science and Technology
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    • v.63 no.6
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    • pp.1423-1432
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    • 2021
  • To elucidate the role and mechanism of microbes, we combined culture-dependent and culture-independent approaches to investigate differences in gut bacterial composition between sows and weaned pigs. Under anaerobic conditions, several nonselective and selective media were used for isolation from fecal samples. All isolated bacteria were identified and classified through 16S rRNA sequencing, and the microbiota composition of the fecal samples was analyzed by metagenomics using next generation sequencing (NGS) technology. A total of 278 and 149 colonies were acquired from the sow and weaned pig fecal samples, respectively. Culturomics analysis revealed that diverse bacterial genus and species belonged to Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes were isolated from sow and weaned pigs. When comparing culture-dependent and culture-independent analyses, 191 bacterial species and 2 archaeal bacterial species were detected through culture-independent analysis, and a total of 23 bacteria were isolated through a culture-dependent approach, of which 65% were not detected by metagenomics. In conclusion, culturomics and metagenomics should be properly combined to fully understand the intestinal microbiota, and livestock-derived microbial resources should be informed by culturomic approaches to understand and utilize the mechanism of host-microbe interactions.

Characterization of beer-spoilage microorganism and its rapid detection by specific PCR primer (맥주오염미생물의 동정과 specific PCR primer의한 신속한 검출 방법)

  • Lee, Taek-In;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.28 no.A
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    • pp.141-147
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    • 2008
  • Several contaminated bacteria such as Lactobacillus brevis and Pediococcus damnosus in beer production cause beer spoilage by producing off flavours and turbidity. Detection of these organisms is complicated by the strict anaerobic conditions and lengthy incubation times required for their cultivation, consequently there is a need for more rapid detection methods. Recently, two contaminated strains were isolated from vessel of beer production and identified as Lactobacillus species by API kit identificaton as well as 16S-23S ITS sequencing analyses. Two isolated strains were named as Lactobacillus sp. HLA1 and Lactobacillus HLB2, respectively. A polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Lactobacillus sp.. Two sets of primer pairs (HLA1-F/HLA1-R and HLB2-F/HLB2-R) were designed for the amplification of a 1576 base pair (bp) fragment of the HLA1 16S-23S rRNA gene and 1888 bp fragement of the HLB2 16S-23S rRNA. Amplified PCR products were highly specific to detect corresponding bacteria when other contaminated strains were used as PCR templates. However, detection of both strains were limited when $100{\mu}{\ell}$ of cultured samples were mixed with $100m{\ell}$ of beer sample in arbitrary manner. The sensitivity of the assay still needs to be improved for direct detection of the small amounts of bacteria present in beer.

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Comparison of Anodic Community in Microbial Fuel Cells with Iron Oxide-Reducing Community

  • Yokoyama, Hiroshi;Ishida, Mitsuyoshi;Yamashita, Takahiro
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.757-762
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    • 2016
  • The group of Fe(III) oxide-reducing bacteria includes exoelectrogenic bacteria, and they possess similar properties of transferring electrons to extracellular insoluble-electron acceptors. The exoelectrogenic bacteria can use the anode in microbial fuel cells (MFCs) as the terminal electron acceptor in anaerobic acetate oxidation. In the present study, the anodic community was compared with the community using Fe(III) oxide (ferrihydrite) as the electron acceptor coupled with acetate oxidation. To precisely analyze the structures, the community was established by enrichment cultures using the same inoculum used for the MFCs. High-throughput sequencing of the 16S rRNA gene revealed considerable differences between the structure of the anodic communities and that of the Fe(III) oxide-reducing community. Geobacter species were predominantly detected (>46%) in the anodic communities. In contrast, Pseudomonas (70%) and Desulfosporosinus (16%) were predominant in the Fe(III) oxide-reducing community. These results demonstrated that Geobacter species are the most specialized among Fe(III)-reducing bacteria for electron transfer to the anode in MFCs. In addition, the present study indicates the presence of a novel lineage of bacteria in the genus Pseudomonas that highly prefers ferrihydrite as the terminal electron acceptor in acetate oxidation.

Multiple liver infection by Sphingomonas sp. and Bacillus sp. in a wild rodent (Apodemus agrarius): a case report

  • Kim, Jiro;Monoldorova, Sezim;Kim, Soojin;Kim, Nam Kyung;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.71-74
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    • 2017
  • The first case of liver infection caused by Sphingomonas sp. and Bacillus sp. in a wild rodent is reported. A captured wild rodent, Apodemus agrarius (A. agrarius), presented with multiple liver abscess-like nodules (diameter 0.7~2.4 mm) in which Gram-positive and Gram-negative bacilli were detected simultaneously. These were grown in aerobic and anaerobic cultures, respectively, and were identified as Sphingomonas sp. and Bacillus sp., respectively, according to 16S rRNA sequencing.