| Project/Area Number |
02558009
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory animal science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MITSUOKA Tomotari The Univ.of Tokyo, Fac.of Agricul., Honorary Prof., 農学部, 名誉教授 (30157549)
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| Co-Investigator(Kenkyū-buntansha) |
MAEJIMA Kazuyoshi Keio Univ., Sch.of Med., Prof., 医学部, 教授 (70051464)
GOTO Naoaki The Univ.of Tokyo, Fac.of Agricul., Prof., 農学部, 教授 (70011989)
HIRAYAMA Kazuhiro The Univ.of Tokyo, Fac.of Agricul., Assis.Prof., 農学部, 助手 (60208858)
ITOH Kikuji The Univ.of Tokyo, Fac.of Agricul., Assoc.Prof., 農学部, 助教授 (50100045)
TAKAHASHI Eiji The Univ.of Tokyo, Fac.of Agricul., Prof., 農学部, 教授 (50183439)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥18,200,000 (Direct Cost: ¥18,200,000)
Fiscal Year 1992: ¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 1991: ¥5,500,000 (Direct Cost: ¥5,500,000)
Fiscal Year 1990: ¥8,900,000 (Direct Cost: ¥8,900,000)
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| Keywords | SPF mouse / SPF rat / Gnotobiote / Intestinal flora / ノトバイオ-ト / 腸内フロ-ラ / <Staphylococcus>___ー <qureus>___ー |
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| Research Abstract |
The purpose of this project is first of all to produce of gnotobiotic mice and rats with normal physiological function for standardizing the intestinal flora of SPF mice and rats, and then to develop the preservation and monitoring system of standardized intestinal flora of mice and rats. Our results were as follows :
1. Gnotobiotic (GB) rats with normal physiological function were produced by using the methods which already have established for production of gnotobiotic mice. Namely, 3 strains of Lactobachillus (L.acidophilus, L.murinus, L.salivarius) and 28 strains of Bacteroidaceae isolated from feces of conventional (CV) rats, and chloroform-treated CV rat feces (survive only clostridia) were inoculated into germfree (GF) rats. These ex-Gf rats were free from Clostridium difficile and eliminated Pseudomonas aeruginosa inoculated orally.
2. Gb rat flora was kept in GF mice inoculated with fecal suspension of GB rats. The production of GB rats was possible only by oral inoculation of feces of GB rats and rat-flora-mice.
3. Standardized mouse flora kept in the intestine of GB mice were inoculated into 4 different strains of GF mice (CF#1, IQI,NC,BALB/c). There were no differences in cecal size and the composition of fecal flora among GF mouse strains.
4. In SPF rat colony established by transferring GB rats, composition of their fecal flora was kept stably in the colony.
5. Preservation method for standardized mouse flora by freezing was examined. Finally, in 1 : 100 fecal suspension in nutrient broth with 10 or 20% glycerol kept in -80 C floral bacteria were kept for 1 year. By inoculation with preserved floral suspension into GF mice, GB mice with normal physiological function were able to be produced, and their floral composition was similar to original GB mice more than that kept in GB mouse intestine for 1 year.
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