• 제목/요약/키워드: 16S-rRNA

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Investigation of Bacterial Diversity in Membrane Bioreactor and Conventional Activated Sludge Processes from Petroleum Refineries Using Phylogenetic and Statistical Approaches

  • Silva, Cynthia;Jesus, Ederson C.;Torres, Ana P. R.;Sousa, Maira P.;Santiago, Vania M. J.;Oliveira, Valeria M.
    • Journal of Microbiology and Biotechnology
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    • 제20권3호
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    • pp.447-459
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    • 2010
  • Bacterial diversity of two distinct wastewater treatment systems, conventional activated sludge (CAS) and membrane bioreactor (MBR), of petroleum refineries were investigated through 16S rRNA gene libraries. Sequencing and phylogenetic analysis showed that the bacterial community composition of sludge samples was distinct between the two wastewater treatment systems. MBR clones belonged predominantly to Class Betaproteobacteria, represented mainly by genera Thiobacillus and Thauera, whereas CAS clones were mostly related to Class Alphaproteobacteria, represented by uncultured bacteria related to Order Parvularculales. Richness estimators ACE and Chao revealed that the diversity observed in both libraries at the species level is an underestimate of the total bacterial diversity present in the environment and further sampling would yield an increased observed diversity. Shannon and Simpson diversity indices were different between the libraries and revealed greater bacterial diversity for the MBR library, considering an evolutionary distance of 0.03. LIBSHUFF analyses revealed that MBR and CAS communities were significantly different at the 95% confidence level ($P{\leq}0.05$) for distances $0{\leq}D{\leq}0.20$. This work described, qualitatively and quantitatively, the structure of bacterial communities in industrial-scale MBR and CAS processes of the wastewater treatment system from petroleum refineries and demonstrated clearly differentiated communities responsible for the stable performance of wastewater treatment plants.

Occurrence of Bacterial Stem Rot of Ranunculus asiaticus Caused by Pseudomonas marginalis in Korea

  • Li, Weilan;Ten, Leonid N.;Kim, Seung-Han;Lee, Seung-Yeol;Jung, Hee-Young
    • Research in Plant Disease
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    • 제24권2호
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    • pp.138-144
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    • 2018
  • In December 2016, stem rot symptoms were observed on Persian buttercup (Ranunculus asiaticus) plants in Chilgok, Gyeongbuk, Korea. In the early stage of the disease, several black spots appeared on the stem of infected plants. As the disease progressed, the infected stem cleaved and wilted. The causal agent was isolated from a lesion and incubated on Reasoner's 2A (R2A) agar at $25^{\circ}C$. Total genomic DNA was extracted for phylogenetic analysis. Based on the 16S rRNA gene analysis, the isolated strain was found to belong to the genus Pseudomonas. To identify the isolated bacterial strain at the species level, the nucleotide sequences of the gyrase B (gyrB) and RNA polymerase D (rpoD) genes were obtained and compared with the sequences in the GenBank database. As the result, the causal agent of the stem rot disease was identified as Pseudomonas marginalis. To determine the pathogenicity of the isolated bacterial strain, it was inoculated into the stem of healthy R. asiaticus plant, the inoculated plant showed a lesion with the same characteristics as the naturally infected plant. Based on these results, this is the first report of bacterial stem rot on R. asiaticus caused by P. marginalis in Korea.

Identification of Non-Aggregatibacter actinomycetemcomitans Bacteria Grown on the Tryptic soy-Serum-Bacitracin-Vancomycin Medium

  • Jo, Eojin;Park, Soon-Nang;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • 제41권4호
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    • pp.199-208
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    • 2016
  • The aim of this study was to identify the non-Aggregatibacter actinomycetemcomitans bacteria grown on the tryptic soy-serum-bacitracin-vancomycin (TSBV) medium, an A. actinomycetemcomitans selective medium. A total of 82 unidentified bacterial isolates from the oral cavities of a Korean population were kindly provide by the Korean Collection for Oral Microbiology. All the clinical isolates were grown on TSBV medium and bacterial DNA purified from each isolate was subjected to PCR with universal primers specific for bacterial 16S rRNA genes (16S rDNAs) sequence. The each bacterial 16S rDNA was amplified by PCR and the nucleotide sequences of it was determined by the dideoxynucleotide chain termination method. They were identified by 16S rDNA sequence comparison method at the specie-level. The data showed that Neisseria spp. (42 strains), Fusobacterium spp. (10 strains), Capnocytophaga spp. (8 strains), Propionibacterium acnes (5 strains), Aggregatibacter aprophilus (4 strains), Campylobacter spp. (5 strains), Veillonella dispar (3 strains), Streptococcus sp. (1 strain), Haemophilus parainfluenzae (1 strain), Leptotrichia wadei (1 strain), Morococcus sp./Neisseria sp. (1 strain), and Staphylococcus sp. (1 strain) were identified. These results could be used to develop a new A. actinomycetemcomitans-selective medium which is more effective than the TSBV medium in future studies.

The complete mitochondrial genome sequence of the indigenous I pig (Sus scrofa) in Vietnam

  • Nguyen, Hieu Duc;Bui, Tuan Anh;Nguyen, Phuong Thanh;Kim, Oanh Thi Phuong;Vo, Thuy Thi Bich
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권7호
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    • pp.930-937
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    • 2017
  • Objective: The I pig is a long nurtured longstanding breed in Vietnam, and contains excellent indigenous genetic resources. However, after 1970s, I pig breeds have become a small population because of decreasing farming areas and increasing pressure from foreign breeds with a high growth rate. Thus, there is now the risk of the disappearance of the I pigs breed. The aim of this study was to focus on classifying and identifying the I pig genetic origin and supplying molecular makers for conservation activities. Methods: This study sequenced the complete mitochondrial genome and used the sequencing result to analyze the phylogenetic relationship of I pig with Asian and European domestic pigs and wild boars. The full sequence was annotated and predicted the secondary tRNA. Results: The total length of I pig mitochondrial genome (accession number KX094894) was 16,731 base pairs, comprised two rRNA (12S and 16S), 22 tRNA and 13 mRNA genes. The annotation structures were not different from other pig breeds. Some component indexes as AT content, GC, and AT skew were counted, in which AT content (60.09%) was smaller than other pigs. We built the phylogenetic trees from full sequence and D loop sequence using Bayesian method. The result showed that I pig, Banna mini, wild boar (WB) Vietnam and WB Hainan or WB Korea, WB Japan were a cluster. They were a group within the Asian clade distinct from Chinese pigs and other Asian breeds in both phylogenetic trees (0.0004 and 0.0057, respectively). Conclusion: These results were similar to previous phylogenic study in Vietnamese pig and showed the genetic distinctness of I pig with other Asian domestic pigs.

A report of eight unrecorded radiation resistant bacterial species in Korea isolated in 2018

  • Jang, Jun Hwee;Sathiyaraj, Gayathri;Sathiyaraj, Srinivasan;Lee, Jin Woo;Kim, Ju-Young;Maeng, Soohyun;Lee, Ki-Eun;Lee, Eun young;Kang, Myung Suk;Kim, Myung Kyum
    • Journal of Species Research
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    • 제7권3호
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    • pp.210-221
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    • 2018
  • Eight bacterial strains assigned to the phylum Firmicutes were isolated from the soil samples in Korea. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains 18JY14-16, 18JY14-35, 18JY42-5, 18JY12-20, 18JY35-8, 18JY76-9, 18JY39-1 and 18JY54-12 were most closely related to Paenibacillus lupini (MH497638; 99.4%), Paenibacillus illinoisensis (MH497643; 99.8%), Paenibacillus tundrae (MH497658; 99.7%), Paenibacillus selenitireducens (MH497639; 99.4%), Paenibacillus eucommiae (MH 497640; 99.9%), Paenibacillus vini (MH497654; 99.4%), Paenibacillus gorillae (MH497647; 99.5%), and Paenibacillus macquariensis (MH497649; 99.9%) respectively. These Paenibacillus species were Gram-stain-positive, rod-shaped and radiation resistant bacteria. This is the first report of these nine bacterial species in Korea.

Characterization of Korean Erwinia carotovora Strains from Potato and Chinese Cabbage

  • Seo, Sang-Tae;Koo, Jun-Hak;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Plant Pathology Journal
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    • 제20권4호
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    • pp.283-288
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    • 2004
  • Four Erwinia carotovora strains isolated from potatoes showing blackleg symptoms and rotted Chinese cabbage were analysed by biochemical tests and sequence analysis of 16S rDNA and 16S-23S rRNA intergenic spacer (IGS) regions, and the data were compared to related E. carotovora strains. Based on the results of the biochemical tests and sequence analysis, 2 of the 4 strains were identified as E. carotovora subsp. carotovora (Ecc), whereas the rest strains were distinct from Ecc. The last two strains, HCC3 and JEJU, were biochemically similar to E, carotovora subsp. atroseptica (Eca). However, the results of sequence analysis and Eca-specific PCR assays showed that the strains were distinct from Eca. On the basis of 16S rDNA sequence analysis, HCC3 and JEJU strains were placed in E. carotovora subsp. odorifera and E. carotovora subsp. wasabiae, respectively. The results of sequence analysis and specific PCR assay for Eca indicated that Asian Eca strains were distinct from European Eca strains, although they were phenotycally homogeneous.

Characteristics of Histamine Forming Bacteria from Tuna Fish Waste in Korea (국내 참치 부산물 내 히스타민 생성 주요 세균의 특성 구명)

  • Bang, Min-Woo;Chung, Chang-Dae;Kim, Seon-Ho;Chang, Moon-Baek;Lee, Sung-Sil;Lee, Sang-Suk
    • Journal of Life Science
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    • 제19권2호
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    • pp.277-283
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    • 2009
  • Biogenic amines are generally formed through the decarboxylation of specific free amino acids by exogenous decarboxylases released by microbial species associated with the fish products and fermented feeds. This study was conducted to investigate the properties of e tuna waste regarding the control of degradation of biogenic amines (histamine, tyramine, tryptamine, putrescine, and cadaverine) that might be related with the anti-nutritional factor of the tuna waste that is used for manufacturing domestic fish meal. The values of pH and the salt content were 6.51, 3.35% in tuna waste and 5.58 and 5.83% in tuna fish meal, respectively. The strains and dominant bacteria tested in the tuna waste sample were 9.20, 9.29, 5.67, 7.82 and 7.58 log CFU/g of total bacteria, aerobic plate count (APC), total coliform (TC), Lactobacillus spp. and Bacillus spp., respectively. The main histamine forming-bacteria (HFB) in tuna waste were detected by silica gel thin-layer chromatography (TLC) and 7 histamine-forming bacterial species were isolated among microbes grown in selective medium. The histamine concentration was determined by detection of fluorescence of ο-phthaldialdehyde (OPA) derivatives using HPLC and the date were used to reconfirm the identities of the amine-producing bacteria. The 15 histamine- forming bacteria strains grown in trypicase soy broth (TSB) supplemented with 1% L-histidine (TSBH) were identified as Lactococcus(L.) lactis subsp. lactis, Klebsiella pneummonlae, L. garvieae 36, Vibrio olivaceus, Hafnia alvei and L. garvieae which were main dominant amine - producing strains, and Morganella morganii identified by 16S ribosomal RNA (rRNA) sequencing with PCR amplification. A Phylogenetic tree generated from the 16S rRNA sequencing data showed different phyletic lines that could be readily classified as biogenic amine forming gram-positive and negative bacteria.

Complete Mitochondrial Genome Sequence and Genetic Diversity of Duroc Breed (돼지 Duroc 품종에서 미토콘드리아 유전체 서열의 특성과 집단의 유전적 다양성)

  • Cho, 1.C.;Han, S.H.;Choi, Y.L.;Ko, M.S.;Lee, J.G;Lee, J.H;Jeon, J .T
    • Journal of Animal Science and Technology
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    • 제46권6호
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    • pp.937-946
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    • 2004
  • Duroc is widely used to improve the meat quality and productivity. To elucidate the phylogenetic relation and the sequence specificity for the maternal property, the complete sequence of mitochondrial genome was determined and the population diversity of Duroc was investigated in this study. The length of mtDNA tested is 16,584-bp. There are several insertion/deletion mutations in the control region and coding regions for tRNA and rRNA, respectively, but not in peptide-coding regions. Four peptide-coding genes(COⅡ, COⅢ, ND3 and ND4) showed incomplete termination codon sequences such as T--, and two(ND2 and ND4L) did alternative initiation codons(AIC), respectively. Especially, the initiation codon sequences of ND2 gene were polymorphic in this population. Polymorphisms were detected in 11-bp duplication motif within control region as well as ND2 and CYTB. Variation patterns observed from the tests on three mtDNA regions were linked completely and then two haplotypes obtained from combining the data dividing this population. Duroc mtDNA is observed at the European pig cluster in the phylogenetic tree, however, the results from the population analyses supported previous opinions. This study suggests that the breed Duroc was mainly originated from the European pig lineage, and Asian lineage was also used to form the pig breed Duroc as maternal progenitors.

Development and Application of Detection Method for Aphanizomenon flos-aquae not Usable as a Food Materials in Korea (식품 중 사용금지 원료인 Aphanizomenon flos-aquae 검출법 개발 및 응용)

  • Park, Yong-Chjun;Shin, Seung-Jung;Lee, Ho-Yeon;Kim, Yong-Sang;Kim, Mi-Ra;Lee, Sang-Jae;Lee, Hwa-Jung
    • Journal of Food Hygiene and Safety
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    • 제28권2호
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    • pp.188-193
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    • 2013
  • Anatoxin-a, saxitoxin and neosaxitoxin are produced by Aphanizomenon flos-aquae that is a sort of the cyanobacteria phylum. Therefore, it is not permitted for food materials in Korea. Traditionally, the classification of cyanobacteria has been based on morphological characters such as trichome width, cell size, division planes, shape, and the presence of character such as gas vacuole. But, some diagnostic features, such as gas vacuole or akinetes, can show variation with different environmental or growth conditions and even be lost during cultivation. Therefore, we developed detection method for functional foods containing Aph. flos-aquae by PCR. To design the primer, 16S rRNA region of Aph. flos-aquae, Spirulina laxissima, and Spirulina spp. registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for comparative analysis, BioEdit ver. 7.0.9.0. was used. As a result, we was design AFA-F1/AFA-R1 (363 bp) primer for the differentiation Aph. flos-aquae from chlorella, spirulina, green tea, and spinach. Also, it could be distinguished chlorella and spirulina products those are made to contain 1% Aph. flos-aquae.

Antimicrobial activities of Burkholderia sp. strains and optimization of culture conditions (Burkholderia sp. OS17의 항균활성 증진을 위한 배양최적화)

  • Nam, Young Ho;Choi, Ahyoung;Hwang, Buyng Su;Chung, Eu Jin
    • Korean Journal of Microbiology
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    • 제54권4호
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    • pp.428-435
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    • 2018
  • In this study, we isolated and identified bacteria from freshwater and soil collected from Osang reservoir, to screen antimicrobial bacteria against various pathogenic bacteria. 38 strains were isolated and assigned to the class Proteobacteria (22 strains), Actinobacteria (7 strains), Bacteroidets (6 strains), and Firmicutes (3 strains) based on 16S rRNA gene sequence analysis. Among them, strain OS17 showed a good growth inhibition against 5 methicillin-resistant Staphylococcus aureus subsp. aureus strains and Bacillus cereus, Bacillus subtilis, Filobasidium neoformans. As a result of the 16S rRNA gene sequence analysis, strain OS17 show the high similarity with Burkholderia ambifaria $AMMD^T$, B. diffusa $AM747629^T$, B. tettitorii $LK023503^T$ 99.8%, 99.7%, 99.6%, respectively. We investigated cell growth and antimicrobial activity according to commercial culture medium, temperature, pH for culture optimization of strain OS17. Optimal conditions for growth and antimicrobial activity in strain OS17 were found to be: YPD medium, $35^{\circ}C$ and pH 6.5. When the strain was cultured in LB, NB, TSB, R2A media at $20^{\circ}C$ and $25^{\circ}C$, the antimicrobial activity did not show. Culture filtrate of strain OS17 showed antimicrobial activity against 5 MRSA strains, Bacillus cereus, Bacillus subtilis, and Filobasidium neoformans with inhibition zones from 2 to 8 mm. Optimal reaction time was 48 h in YPD medium, 100 rpm and 0.3 vvm in 2 L-scale fed-batch fermentation process for antimicrobial activity. Culture optimization of strain OS17 can be improved on antimicrobial activity. Therefore, the antimicrobial activity of Burkholderia sp. OS17 had potential as antibiotics for pathogens including MRSA.