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      KCI등재 SCIE SCOPUS

      Molecular Analysis of Colonized Bacteria in a Human Newborn Infant Gut

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      https://www.riss.kr/link?id=A104675457

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      다국어 초록 (Multilingual Abstract)

      The complex ecosystem of intestinal microflora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut.
      To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA isolated
      from feces on the 1st, 3rd, and 6th day.
      16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and 50oC annealing temperature.
      Nine independent libraries were produced
      by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones (76.7%) of all 325 isolated clones
      were characterized as known species, while other 105 clones (32.3%) were characterized as unknown species.
      The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days.
      Thus, set A primers were better suited for this type of molecular ecological analysis.
      On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present.
      The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and treptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present.
      The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.
      번역하기

      The complex ecosystem of intestinal microflora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fra...

      The complex ecosystem of intestinal microflora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut.
      To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA isolated
      from feces on the 1st, 3rd, and 6th day.
      16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and 50oC annealing temperature.
      Nine independent libraries were produced
      by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones (76.7%) of all 325 isolated clones
      were characterized as known species, while other 105 clones (32.3%) were characterized as unknown species.
      The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days.
      Thus, set A primers were better suited for this type of molecular ecological analysis.
      On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present.
      The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and treptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present.
      The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.

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      다국어 초록 (Multilingual Abstract)

      The complex ecosystem of intestinal microflora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut.
      To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA isolated from feces on the 1st, 3rd, and 6th day.
      16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and 50oC annealing temperature.
      Nine independent libraries were produced by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones (76.7%) of all 325 isolated clones were characterized as known species, while other 105 clones (32.3%) were characterized as unknown species.
      The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days.
      Thus, set A primers were better suited for this type of molecular ecological analysis.
      On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present.
      The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and treptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present.
      The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.
      번역하기

      The complex ecosystem of intestinal microflora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fra...

      The complex ecosystem of intestinal microflora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut.
      To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA isolated from feces on the 1st, 3rd, and 6th day.
      16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and 50oC annealing temperature.
      Nine independent libraries were produced by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones (76.7%) of all 325 isolated clones were characterized as known species, while other 105 clones (32.3%) were characterized as unknown species.
      The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days.
      Thus, set A primers were better suited for this type of molecular ecological analysis.
      On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present.
      The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and treptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present.
      The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.

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      참고문헌 (Reference)

      1 Cole, "previewing a new autoaligner that allows regular updates andthe new prokaryotic taxonomy. Nucleic Acids Res. 31" 31 : 442-443, 2003

      2 Hecht, "p.113-119. American Society for Microbiology" 113-119, 2003

      3 Cho, "and Y.K. Choi. 2003. The genetic diversityanalysis of the bacterial community in groundwater by denaturinggradient gel electrophoresis" 41 : 327-334, 2003

      4 Marchesi, "and W.G. Wade. 1998. Design and evaluationof useful bacterium-specific PCR primers that amplifygenes coding for bacterial 16S rRNA. Appl. Environ. Microbiol.64" 64 : 795-799, 1998

      5 R. Chierici, "and V. Vigi. 2003. Intestinalmicroflora in early infancy" 91 : 48-55, 2003

      6 Wang, "and T.H. Florin. 2003.Molecular characterization of the microbial species that colonizehuman ileal and colonic mucosa by using 16S rDNAsequence analysis. J. Appl. Microbiol. 95" 95 : 508-520, 2003

      7 C, "and S. Rolleke. 2001.An advanced molecular strategy to identify bacterial communitieson art objects. J. Microbiol. Methods. 45" 45 : 77-87, 2001

      8 Ward, "and M.M. Bateson. 1990. 16S rRNAsequences reveal numerous uncultured microorganisms in anatural community. Nature 345" 63-65, 1990

      9 C.M. Cavanaugh, "and M.F. Polz. 2002. Sequencingindependentmethod to generate oligonucleotide probes targetinga variable region in bacterial 16S rRNA by PCR withdetachable primers. Appl. Environ. Microbiol. 68" 68 : 6077-6086, 2002

      10 Bonnet, "and M.D. Collins. 2002.Differences in rDNA libraries of faecal bacteria derived from 10-and 25-cycle PCRs. Int. J. Syst. Evol. Microbiol. 52" 52 : 757-763, 2002

      1 Cole, "previewing a new autoaligner that allows regular updates andthe new prokaryotic taxonomy. Nucleic Acids Res. 31" 31 : 442-443, 2003

      2 Hecht, "p.113-119. American Society for Microbiology" 113-119, 2003

      3 Cho, "and Y.K. Choi. 2003. The genetic diversityanalysis of the bacterial community in groundwater by denaturinggradient gel electrophoresis" 41 : 327-334, 2003

      4 Marchesi, "and W.G. Wade. 1998. Design and evaluationof useful bacterium-specific PCR primers that amplifygenes coding for bacterial 16S rRNA. Appl. Environ. Microbiol.64" 64 : 795-799, 1998

      5 R. Chierici, "and V. Vigi. 2003. Intestinalmicroflora in early infancy" 91 : 48-55, 2003

      6 Wang, "and T.H. Florin. 2003.Molecular characterization of the microbial species that colonizehuman ileal and colonic mucosa by using 16S rDNAsequence analysis. J. Appl. Microbiol. 95" 95 : 508-520, 2003

      7 C, "and S. Rolleke. 2001.An advanced molecular strategy to identify bacterial communitieson art objects. J. Microbiol. Methods. 45" 45 : 77-87, 2001

      8 Ward, "and M.M. Bateson. 1990. 16S rRNAsequences reveal numerous uncultured microorganisms in anatural community. Nature 345" 63-65, 1990

      9 C.M. Cavanaugh, "and M.F. Polz. 2002. Sequencingindependentmethod to generate oligonucleotide probes targetinga variable region in bacterial 16S rRNA by PCR withdetachable primers. Appl. Environ. Microbiol. 68" 68 : 6077-6086, 2002

      10 Bonnet, "and M.D. Collins. 2002.Differences in rDNA libraries of faecal bacteria derived from 10-and 25-cycle PCRs. Int. J. Syst. Evol. Microbiol. 52" 52 : 757-763, 2002

      11 Tung, "and K.Y. Sanbonmatsu. 2002. All-atomhomology model of the Escherichia coli 30S ribosomal subunit.Nat. Struct. Biol. 9" 9 : 750-755, 2002

      12 A, "and J. Dore. 1999. Direct analysis of genes encoding 16SrRNA from complex communities reveals many novel molecularspecies within the human gut. Appl. Environ. Microbiol.65" 65 : 4799-4807, 1999

      13 Mackie, "and H.R. Gaskins. 1999. Developmentalmicrobial ecology of the neonatal gastrointestinal tract. Am. J.Clin. Nutr. 69" 69 : 1035S-1045S, 1999

      14 A.J, "and F. Rodriguez-Valera. 1999.Patterns of sequence variation in two regions of the 16S rRNAmultigene family of Escherichia coli. Int. J. Syst. Bacteriol. 49" 49 : 601-610, 1999

      15 Li, "and E.Topp. 2003. Evaluation of QIAamp DNA Stool Mini Kit forecological studies of gut microbiota. J. Microbiol. Methods. 54" 54 : 13-20, 2003

      16 F, "and E. Stackebrandt. 1997.Determination of microbial diversity in environmental samples pitfalls of PCR-based rRNA analysis. FEMS Microbiol. Rev.21" 21 : 213-229, 1997

      17 Kang, "and C.M. Kim. 2004.Web-based research assistant tools for analysis of microbialdiversity. J. Fuzzy Logic and Intelligent Systems. 14" 14 : 545-550, 2004

      18 A.L, "and A.R. Bird. 2002. A comparison offive methods for extraction of bacterial DNA from human faecalsamples. J. Microbiol. Methods. 50" 50 : 131-139, 2002

      19 C.F, "and A.D. Akkermans.2002. Molecular monitoring of succession of bacterial communitiesin human neonates. Appl. Environ. Microbiol. 68" 68 : 219-226, 2002

      20 F, "Malagelada . 2003 Gut flora in health and disease.Lancet. 361" 361 : 512-519, 2003

      21 "J.J. 1999. Influences of microbiota on intestinal immune systemdevelopment. Am. J. Clin. Nutr. 69" 69 : 1046S-1051S, 1999

      22 "J.C. 1998. The value of microbial diversity. Curr.Opin. Microbiol. 1" 1 : 278-285, 1998

      23 Wilson, "I.G. 1997. Inhibition and facilitation of nucleic acid amplification. Appl. Environ. Microbiol. 63 and K. Fujita. 1983. Development and differences of intestinal flora in the neonatal period in breast-fedand bottle-fed infants. Pediatrics 72" 63 : 3741-3751, 1997

      24 Hur, "I. and J. Chun. 2004. A method for comparing multiple bacterialcommunity structures from 16S rDNA clone librarysequences. J. Microbiol. 42" 42 : 9-13, 2004

      25 "I. 2002. Molecular community analysis of microbial diversity.Curr. Opin. Biotechnol. 13" 13 : 213-217, 2002

      26 Tannock, "G.W. 2001 Molecular assessment of intestinal microflora.Am. J. Clin. Nutr. 73" 73 : 410S-414S, 2001

      27 Wang, "G.C. and Y. Wang. 1997. Frequency of formation of chimericmolecules as a consequence of PCR coamplification of 16SrRNA genes from mixed bacterial genomes. Appl. Environ.Microbiol. 63" 63 : 4645-4650, 1997

      28 "Factors affecting PCR-mediated recombination" 4 : 482-486, 2002

      29 Shanahan, "F. 2002. The host-microbe interface within the gut. BestPract. Res. Clin." 16 : 915-931, 2002

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