1 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /,, No.0,,.-,/* (,**/) 243 Lactococcus lactis subsp. cremoris FC * +, *, *, *, * - *. * + * + *, * - *. E#ects of Fermented Milk with Lactococcus lactis subsp. cremoris FC on Defecation Frequency and Fecal Microflora in Healthy Elderly Volunteers Toshiya Toda* +, *,, Hideki Kosaka*,, Masakazu Terai*,, Hideki Mori* -, Yoshimi Benno*. and Yukio Yamori* + * + Research Institute for Production Development, +/ Morimotocho, Shimogamo, Sakyo-ku, Kyoto 0*0 *2*/ *, Fujicco Co., Ltd, 0 +-. Minatojima-nakamachi, Chuo-ku, Kobe, Hyogo 0/* 2//2 * - Institute for Health Restoration, + 2. Higashikawasakicho, Chuo-ku, Kobe, Hyogo 0/* **.. *. Japan Collection of Microorganisms, Riken BioResource Center,, + Hirosawa, Wako, Saitama, -/+ *+32 Caspian Sea Yogurt, viscous fermented milk with Lactococcus lactis subsp. cremoris, has become popular in Japan. Since its health e#ect has not been studied su$ciently for human, the fermented-milk with L. lactis subsp. cremoris FC was tested for the e#ects on defecation frequency, fecal characteristics and fecal microflora in healthy elderly volunteers. 1* volunteers aged in average, 01.-.., and 00.3 /.- years old in males and females, respectively, were randomized into, groups for double-blind placebo-controlled crossover study. +/* g of fermented milk drink containing,* soybean milk ( +./ +* 3 CFU/g of L. lactis subsp. cremoris FC, and -.+ +* 1 CFU/g of Streptococcus salivarius subsp. thermophilus) or+/* g of fermented products without L. lactis subsp. cremoris FC were consumed daily for + month test and control periods, between which there was + month washout period. The defecation frequency and fecal characteristics were observed during these test, control and washout periods. The fecal microflora, ammonia concentration were examined in 3 subjects. The volume of defecation was significantly increased, weeks after the administration in both groups. The frequency and number of days of defecation were significantly increased only in the test group. The fecal shape and color were improved in both groups particularly in costive subjects. The ratio of Bifidobacteria to total bacteria increased and the frequency of occurrence of Clostridium perfringens tended to decrease in the test group, but there was no change in the control group. The fecal ammonia concentration also decreased in the test group. (Received Jul. 3,,**. ; Accepted Mar. 3,,**/),* Metchniccoff + Lactobacillus Bifidobacterium * + *, * - *. 0*0 *2*/ +/ 0/* 2//2 0 +-. 0/* **.. + 2. / 0 -/+ *+32, + Corresponding author ttoda@fujicco.co.jp,. / Lactococcus lactis subsp. cremoris 0 Acetobacter orientalis A. orientalis L. lactis subsp. cremoris
244 /, 0,**/ 0 2 + L. lactis subsp. cremoris Lactococcus lactis subsp cremoris 1 Lactococcus Lactococcus 2 3 L. lactis subsp. cremoris +* Lactococcus lactis subsp. cremoris L. lactis subsp. cremoris + +320,**, L. lactis subsp. cremoris FC 0 1 L. lactis subsp. cremoris FC Streptococcus salivarius subsp. thermophilus /+* +,,* 2.+ L. lactis subsp. cremoris FC +./ +* 3 CFU g S. salivarius subsp. thermophilus /+* -.+ +* 1 CFU g L. lactis subsp. cremoris FC S. salivarius subsp. thermophilus /+* S. salivarius subsp. thermophilus /+* 1.* +* 1 CFU g, -, -2 01.+..2 - + 1*, A, B + A B + A B + +. + +/* g, + +/* g. +, + +,* cm
3 : FC 245 + :, : - :. : / : / DIC +1 + : DIC,*0, : DIC,.* - : DIC -,+. : DIC -*2 / : DIC -++ + :, : - :. : / : / + :, : - :. : / : / + / / 1* 3 02.2..3 ph, +. + 2* 0 ++ +, +* -/, - -/ - / + g 1 ph +** ph ph 2 Wilcoxon ph t SPSS for Windows ver. +*.* SPSS Inc. +, 1* +/ +, 10.+ 23.2 +-.2 0.* +*.+..,, 2+ 13.3 +1.- 1.1 +.1 +,.. -.,3.0./.. --.2 2.3 /.,.2 +-./,+.* //.1
246 /, 0,**/ 0 4 + Lactococcus lactis subsp. cremoris FC +, +, +. / 24+ -4..42 +42 24/ -40 /4*,4/ 240-4+* 04-,4.* 24, -4*.4/ +4* 24/ -4. /4* +41 24. -4. /40,4,, 04/ *43** /4- +4-* 04, +4, -43 *41 04+ +4-.4+ +4+ 04, +4-.4- +40 04, +4,.4- +4-04. +4+ /4+ +42-14. /4+ -41 +40 143 /4..4.,40 24- /4**** /4/,4.* 14*.4* -41,4* 142.41**.42-4** 24+.4.** /4/ -4-* + +, + - +,* cm +. n // / + / n +* *:p *4*/ ** : p *4*+ *** : p *4**+, Lactococcus lactis subsp. cremoris FC ** p *.*+ -0., -*.2-3, Bifidobacterium Clostoridium perfringens. ph, ph - Lactobacillus Bifidobacterium,.
ῌ³ : ¹Dº»2 FC => HI ¼ 5 - Lactococcus lactis subsp. cremoris FC ± Lactococcus!"#$% & µcz ². ³. Lactococcus lactis subsp. cremoris FC \ 247 \ [ A - ), jkl ] #* P mk `anop A jq\:rs#tk_ uv '( ) * +,+-. /0 123456789 w * + L. lactis subsp. cremoris :;<=>?@ x^ yz{ }~ l Y) : A - BCDE8 FGHI JKL MN+ #*. A Y),[- ]^_ O - P$% 123456789:QRST # )#O #+- +. * PmK x_ + L. lactis subsp. cremoris U9 V"#FG ^ 01.+..2 HI:WXY Z[, "#O :@Y + - F\)] ^_ L. lactis subsp. cremoris FC :;<=> :`a b-. c de+ * Lactococcus - f,[ gh gi # - X x+ #FG )tk_ #*. `an AT# A -. 0.+ & 0., ῌ - 2.+ & 2., ῌ ""( - ª ) «:jw * & - + o
248 /, 0,**/ 0 6 / Lactococcus lactis subsp. cremoris FC ** p *.**+, Lactococcus lactis subsp. cremoris FC Total aerobes Enterobacteriaceae 24+ *4, 142 *4. 24+ *40 14- *4/ 24, *41 14, +40 140 *41 04/ +4* Total anaerobes Bifidobacterium Bifidobacterium Lactobacillus Clostoridium perfringens Clostoridium others Bacteroidaceae Eubacterium +*40 *4. 341 *4/,*4- +.42 04.,4/ /4+ +42 04. +4/ 343 +4* 341 *40 23 +*40 *4- +*4* *40-04, +24/** 04+ +40 /4+ +4-14* *41 +*4+ *4-34. *40 /0 +*4/ *4. 341 *42,-4/,+4, /4/ *43.4/ +4-14- +4* +*4+ *4. 34/ *40 /0 +*4. *4. 34/ *43,24/,/42 04* +4/.43-4+ 041 *42 +*4+ *40 34, *40 01 Total counts +*40 *4. +*40 *4- +*4/ *4. +*4. *4. + g n 3 ** : p *4*+ / +* + +/* g, + +, +/ L. lactis subsp. cremoris FC ph
7 : FC 249 - Lactococcus lactis subsp. cremoris FC ph, ph mg/g 1.4/ 043 04.1 *4-3.20,32 1/4* 342 040* *4.- -21,.- 1.41 043 040, *4-0 -.3,,2 1/4/ 143 04// *4.*..,,-+ n 3 -. +3 2 L. lactis subsp. cremoris FC Bifidobacterium C. perfringens, Enterobacteriaceae Lactobacillus +0 +1 Bifidobacterium,*.-,-./ +** L. lactis subsp. cremoris FC Bifidobacterium Bifidobacterium +2 +3,*,+,,,- C. perfringens C. perfringens L. lactis subsp. cremoris FC Lactococcus Caco, ph 2 Lactococcus 3 L. lactis subsp. cremoris,. L. lactis subsp. cremoris FC L. lactis subsp. cremoris FC L. lactis subsp. cremoris FC L. lactis subsp. cremoris FC + +/* g L. lactis subsp. cremoris FC +./ +* 3 CFU g S. salivarius subsp. thermophilus /+* -.+ +* 1 CFU g,* 1* 01.+..2, + L. lactis subsp. cremoris FC +/* g +, Bifidobacterium Clostridium perfringens + Metchnico#, E., The Prolongation of Life. GP Putman, New York +3*1, Lactobacillus GG 2,+,2 +33. - Spanhaak. S., Havenaar, R. and Schaafsma, G., The e#ect of consumption of milk fermented by Lactobacillus casei strain Shirota on the intestinal microflora and immune parameters in humans. Eur. J. Clin. Nutr., /,, 233 3*1 +332. Link-Amster, H., Rochat, F., Saudan, K.Y., Mignot, O. and Aeschlimann, J.M., Modulation of a specific humoral immune response and changes in intestinal flora mediated through fermented milk intake. FEMS Immunol. Med. Microbiol., +*, // 0- +33.
250 /, 0,**/ 0 8 / +333 0 +- 0-,**, 1 Lactococcus lactis subsp. cremoris F Acetobacter orientalis F +/ p. 2,**, 2 Kimoto, H., Kurisaki, J., Tsuji, N. M., Ohmomo, S. and Okamoto, T., Lactococci as probiotic strain : adhesion to human enterocyte like Caco-, cells and tolerance to low ph and bile. Lett. Appl. Microbiol.,,3, -+- -+0 +333 3 Kimoto, H., Nomura, M., Kobayashi, M., Mizumachi, K. and Okamoto, T., Survival of lactococci during passage through mouse digestive tract. Can. J. Microbiol.,.3, 1*1 1++,**- +* Nakajima, H., Suzuki, Y., Kaizu, H. and Hirota, T., Cholesterol lowering activity of ropy fermented milk. J. Food Sci., /1, +-,1 +-,3 +33, ++ +32* +, - +32, +- Geriatr. Med., ++, -3, -31 +31- +. 1/,,,3,,-- +332 +/ +33* +0 Mitsuoka, T. and Hayakawa, K., Die faekalflora bei menschen. I. Mitteilung : die zusammensetzung der faekalflora der verschiedenen altersgruppen. Zentralbl. Mikrobio. Hyg. I Abt. Orig., A,,-, --- -., +31, +1 +. /2+ /21 +32/ +2 -. -13-21 +312 +3 Benno, Y. and Mitsuoka, T., Impact of Bifidobacterium longum on human fecal microflora. Microbiol. Immunol., -0, 02-03. +33,,* Ueda, K., Immunity provided by colonized enteric bacteria. Bifidobacteria Microflora., /, 01 1, +320,+ Yasui, H. and Ohwaki, M., Enhancement of immune response in peyer s patch cells cultured with Bifidobacterium breve. J. Dairy Sci., 1., ++21 ++3/ +33+,, Kohwi, Y., Imai, K., Tamura, Z. and Hashimoto, Y., Antitummor e#ect of Bifidobacterium infantis in Mice. Gann, 03, 0+- 0+2 +312,- Tsuyuki, S., Yamazaki, S., Akashiba, H., Kamimura, H., Skine, K., Toida, T., Saito, M., Kawashima, T. and Ueda, K., Tumor suppressive e#ect of a cell wall preparation, WPG, from Bifidobacterium infantis in germfree and flora-bearing mice. Bifidobacteria Microflora +*,.- /, +33+,. Ruijssenaars, H.J., Stingele, F. and Hartmans, S., Biodegradability of food-associated extracellular polysaccharides. Curr. Microbaiol.,.*, +3. +33,*** +0 1 3 +1-3