Journal of Microbiology

  • 제43권4호
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  • Pages.345-353
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  • 2005
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  • 1225-8873(pISSN)
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  • 1976-3794(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Molecular Analysis of Colonized Bacteria in a Human Newborn Infant Gut

  • Park Hee-Kyung (Institute for Genomic Medicine, GENEIN Co., Ltd.) ;
  • Shim Sung-Sub (Departments of Pediatrics, College of Medicine, Pusan National University) ;
  • Kim Su-Yung (Departments of Pediatrics, College of Medicine, Pusan National University) ;
  • Park Jae-Hong (Departments of Pediatrics, College of Medicine, Pusan National University) ;
  • Park Su-Eun (Departments of Pediatrics, College of Medicine, Pusan National University) ;
  • Kim Hak-Jung (Departments of Biochemistry, College of Medicine, Pusan National University) ;
  • Kang Byeong-Chul (Division of Applied Bioengineering, Dongseo University) ;
  • Kim Cheol-Min (Departments of Biochemistry, College of Medicine, Pusan National University)
  • 발행 : 2005.08.01
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초록

The complex ecosystem of intestinal micro flora 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 iso-lated from feces on the 1st, 3rd, and 6th day. 16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and $50^{\circ}C$ 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 Streptococcus 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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