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    Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis Analysis of Bacterial Community Structure in the Food, Intestines, and Feces of Earthworms

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    https://www.riss.kr/link?id=A103418839

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    다국어 초록 (Multilingual Abstract) kakao i 다국어 번역

    The bacterial communities in the food, intestines, and feces of earthworms were investigated by PCR-denaturing gradient gel electrophoresis (DGGE). In this study, PCR-DGGE was optimized by testing 6 universal primer sets for microbial 16S rRNA in 6 pure culture strains of intestinal microbes in earthworms. One primer set effectively amplified 16S rRNA from bacterial populations that were found in the food, intestines, and feces of earthworms. Compared with the reference markers from the pure culture strains, the resulting DGGE profiles contained 28 unique DNA fragments. The dominant microorganisms in the food, intestines,and feces of earthworms included Rhodobacterales bacterium, Fusobacteria, Ferrimonas marina, Aeromonas popoffii, and soil bacteria. Other strains, such as Acinetobacter, Clostridium, and Veillonella, as well as rumen bacteria and uncultured bacteria also were present. These results demonstrated that PCR-DGGE analysis can be used to elucidate bacterial diversity and identify unculturable microorganisms.
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    The bacterial communities in the food, intestines, and feces of earthworms were investigated by PCR-denaturing gradient gel electrophoresis (DGGE). In this study, PCR-DGGE was optimized by testing 6 universal primer sets for microbial 16S rRNA in 6 pu...

    The bacterial communities in the food, intestines, and feces of earthworms were investigated by PCR-denaturing gradient gel electrophoresis (DGGE). In this study, PCR-DGGE was optimized by testing 6 universal primer sets for microbial 16S rRNA in 6 pure culture strains of intestinal microbes in earthworms. One primer set effectively amplified 16S rRNA from bacterial populations that were found in the food, intestines, and feces of earthworms. Compared with the reference markers from the pure culture strains, the resulting DGGE profiles contained 28 unique DNA fragments. The dominant microorganisms in the food, intestines,and feces of earthworms included Rhodobacterales bacterium, Fusobacteria, Ferrimonas marina, Aeromonas popoffii, and soil bacteria. Other strains, such as Acinetobacter, Clostridium, and Veillonella, as well as rumen bacteria and uncultured bacteria also were present. These results demonstrated that PCR-DGGE analysis can be used to elucidate bacterial diversity and identify unculturable microorganisms.

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    참고문헌 (Reference)

    1 정영률, "훼손된 산불토양의 미생물다양성" 유기성자원학회 13 (13): 85-90, 2005

    2 Ji, N, "Universal primer PCR with DGGE for rapid detection of bacterial pathogens" 57 : 409-413, 2004

    3 Cocolin, L, "The late blowing in cheese: a new molecular approach based on PCR and DGGE to study the microbial ecology of the alteration process" 90 : 83-91, 2004

    4 Clegg, C.D, "The impact of grassland management regime on the community structure of selected bacterial groups in soils" 43 : 263-270, 2003

    5 Rondon, M.R, "The earth’s bounty: assessing and assessing soil microbial diversity" 17 : 403-409, 1999

    6 Hong-BumCho, "The Genetic Diversity Analysis of the Bacterial Community in Groundwater by Denaturing Gradient Gel Electrophoresis" 한국미생물학회 41 (41): 327-334, 2003

    7 Lane, D.J, "Rapid determination of 16S ribosomal RNA sequences for phylogenetic analysis" 82 : 6955-6959, 1985

    8 Muyzer, G, "Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA" 59 : 695-700, 1993

    9 Horn, M.A, "Nitrous oxide reductase genes (nosZ) of denitrifying microbial populations in soil and the earthworm gut are phylogenetically similar" 72 : 1019-1026, 2006

    10 Ihssen, J, "N2O-producing microorganisms in the gut of the earthworm Aporrectodea caliginosa are indicative of ingested soil bacteria" 69 : 1655-1661, 2003

    1 정영률, "훼손된 산불토양의 미생물다양성" 유기성자원학회 13 (13): 85-90, 2005

    2 Ji, N, "Universal primer PCR with DGGE for rapid detection of bacterial pathogens" 57 : 409-413, 2004

    3 Cocolin, L, "The late blowing in cheese: a new molecular approach based on PCR and DGGE to study the microbial ecology of the alteration process" 90 : 83-91, 2004

    4 Clegg, C.D, "The impact of grassland management regime on the community structure of selected bacterial groups in soils" 43 : 263-270, 2003

    5 Rondon, M.R, "The earth’s bounty: assessing and assessing soil microbial diversity" 17 : 403-409, 1999

    6 Hong-BumCho, "The Genetic Diversity Analysis of the Bacterial Community in Groundwater by Denaturing Gradient Gel Electrophoresis" 한국미생물학회 41 (41): 327-334, 2003

    7 Lane, D.J, "Rapid determination of 16S ribosomal RNA sequences for phylogenetic analysis" 82 : 6955-6959, 1985

    8 Muyzer, G, "Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA" 59 : 695-700, 1993

    9 Horn, M.A, "Nitrous oxide reductase genes (nosZ) of denitrifying microbial populations in soil and the earthworm gut are phylogenetically similar" 72 : 1019-1026, 2006

    10 Ihssen, J, "N2O-producing microorganisms in the gut of the earthworm Aporrectodea caliginosa are indicative of ingested soil bacteria" 69 : 1655-1661, 2003

    11 Tamura, K, "MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0" 24 : 1596-1599, 2007

    12 Endo, A, "Lactobacillus and Bifidobacterium diversity in horse feces, revealed by PCRDGGE" 59 : 651-655, 2009

    13 Spano, G, "In vivo PCR-DGGE analysis of Lactobacillus plantarum and Oenococcus oeni populations in red wine" 54 : 9-13, 2007

    14 Theunissen, J, "Identification of probiotic microorganisms in south African products using PCR-based DGGE analysis" 98 : 11-21, 2005

    15 Walter, J, "Detection and identification of gastrointestinal Lactobacillus species by using denaturing gradient gel electrophoresis and species specific primers" 66 : 297-303, 2000

    16 Miller, K.M, "Denaturing gradient gel electrophoresis (DGGE): a rapid and sensitive technique to screen nucleotide sequence variation in populations" 27 : 1016-1030, 1999

    17 Horn, M.A, "Dechloromonas denitrificans sp. nov., Flavobacterium denitrificans sp. nov., Paenibacillus anaericanus sp. nov. and Paenibacillus terrae strain MH72, N2O-producing bacteria isolated from the gut of the earthworm Aporrectodea caliginosa" 55 : 1255-1265, 2005

    18 Watanabe, T, "DGGE method for analyzing 16S rDNA of methanogenic archaeal community in paddy field soil" 232 : 153-163, 2004

    19 Temmerman, R, "Cultureindependent analysis of probiotic products by denaturing gradient gel electrophoresis" 69 : 220-226, 2003

    20 Rantsiou, K, "Culture-dependent and -independent methods to investigate the microbial ecology of Italian fermented sausages" 71 : 1977-1986, 2005

    21 Li, Z., "Cultivable bacterial community from south China sea sponge as revealed by DGGE fingerprinting and 16S rDNA phylogenetic analysis" 55 : 465-472, 2007

    22 Gurtner, C, "Comparative analyses of the bacterial diversity on two different biodeteriorated wall paintings by DGGE and 16S rDNA sequence analysis" 46 : 229-239, 2000

    23 Liu, W.T, "Characterization of microbial diversity by determining terminal restriction fragment length polymorphisms of genes encoding 16S rRNA" 63 : 4516-4522, 1997

    24 Edwards, C.A, "Biology of earthworms" Chapman Hall 20-45, 1972

    25 Young-Do Nam, "Bacterial, Archaeal, and Eukaryal Diversity in the Intestines of Korean People" 한국미생물학회 46 (46): 491-501, 2008

    26 Fasoli, S, "Bacterial composition of commercial probiotic products as evaluated by PCR-DGGE analysis" 82 : 59-70, 2003

    27 Leung, K, "Bacterial community dynamics in liquid swine manure during storage: molecular analysis using DGGE/ PCR of 16S rDNA" 38 : 169-177, 2001

    28 Zahner, V., "Application of 16S rDNA-DGGE and plate culture to characterization of bacterial communities associated with the sawfly, Acantholyda erythrocephala (Hymenoptera, Pamphiliidae)" 57 : 564-569, 2008

    29 Kwang-Hee Shin, "Analysis of the Anaerobic Bacterial Community in the Earthworm (Eisenia fetida) Intestine" 한국응용생명화학회 47 (47): 147-152, 2004

    30 Kim, T.W, "Analysis of Bacterial and Fungal Communities in Japanese- and Chinese-Fermented Soybean Pastes Using Nested PCR-DGGE" SPRINGER 60 (60): 315-320, 2010

    31 Hyun-Jung Kim, "Analysis of Aerobic and Culturable Bacterial Community Structures in Earthworm (Eisenia fetida) Intestine" 한국응용생명화학회 47 (47): 137-142, 2004

    32 Cocolin, L, "An application of PCR-DGGE analysis to profile the yeast populations in raw milk" 12 : 407-411, 2002

    33 Schabereiter-Gurtner, C, "An advanced molecular strategy to identify bacterial communities on art objects" 45 : 77-87, 2001

    34 Osorio, C.R, "16S rRNA gene sequence analysis of photobacterium damsel and nested PCR method for rapid detection of the causative agent of fish pasteurellosis" 65 : 2942-2946, 1999

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