Objectives: This study aims to establish the baseline profile of intestinal microbiota in pre-school and school-age Japanese children and to investigate the effects of a probiotic on the microbiota. Methods: We analyzed the intestinal microbiota and investigated the effects (before, during and after the ingestion period) on intestinal microbiota and the environment of 6 months of daily ingestion of a probiotic (Lactobacillus casei strain Shirota (LcS)-fermented milk). Results: We performed an open trial in 23 children (14 boys, 9 girls; age 7.7 ± 2.4 years (mean ± SD); BMI 19.6 ± 4.6). The composition of intestinal microbiota of healthy pre-school and school-age children resembled that of adults. During probiotic supplementation, the population levels of Bifidobacterium and total Lactobacillus increased significantly, while those of Enterobacteriaceae, Staphylococcus and Clostridium perfringens decreased significantly. A significant increase in fecal concentrations of organic acids and also a decrease in fecal pH were observed during the ingestion period. However, the patterns of fecal microbiota and intestinal environment were found to revert to the baseline levels (i.e. before ingestion) within 6 months following the cessation of probiotic intake. Conclusion: Regular intake of an LcS-containing probiotic product may modify the gut microbiota composition and intestinal environment in pre-school and school-age children while maintaining the homeostasis of the microbiota.

Keywords:
Fecal microbiota, Lactobacillus casei strain Shirota, Gut microbiota, Organic acids, Pre-school and school-age children , Probiotics
1.
Johnson CL, Versalovic J: The human microbiome and its potential importance to pediatrics. Pediatrics 2012;129:950-960.
2.
Tiihonen K, Ouwehand AC, Rautonen N: Human intestinal microbiota and healthy ageing. Ageing Res Rev 2010;9:107-116.
3.
Parvez S, Malik KA, Ah Kang S, Kim HY: Probiotics and their fermented food products are beneficial for health. J Appl Microbiol 2006;100:1171-1185.
4.
Morais MB, Jacob CM: The role of probiotics and prebiotics in pediatric practice. J Pediatr (Rio J) 2006;82(5 suppl):S189-S197.
5.
Bezirtzoglou E, Stavropoulou E: Immunology and probiotic impact of the newborn and young children intestinal microflora. Anaerobe 2011;17:369-374.
6.
Matsuda K, Tsuji H, Asahara T, Kado Y, Nomoto K: Sensitive quantitative detection of commensal bacteria by rRNA-targeted reverse transcription-PCR. Appl Environ Microbiol 2007;73:32-39.
7.
Matsuda K, Tsuji H, Asahara T, Matsumoto K, Takada T, Nomoto K: Establishment of an analytical system for the human fecal microbiota, based on reverse transcription-quantitative PCR targeting of multicopy rRNA molecules. Appl Environ Microbiol 2009;75:1961-1969.
8.
Fujimoto J, Matsuki T, Sasamoto M, Tomii Y, Watanabe K: Identification and quantification of Lactobacillus casei strain Shirota in human feces with strain-specific primers derived from randomly amplified polymorphic DNA. Int J Food Microbiol 2008;126:210-215.
9.
Jansen GJ, Wildeboer-Veloo AC, Tonk RH, Franks AH, Welling GW: Development and validation of an automated, microscopy based method for enumeration of groups of intestinal bacteria. J Microbiol Methods 1999;37:215-221.
10.
Matsuda K, Tsuji H, Asahara T, Takahashi T, Kubota H, Nagata S, Yamashiro Y, Nomoto K: Sensitive quantification of Clostridium difficile cells by reverse transcription-quantitative PCR targeting rRNA molecules. Appl Environ Microbiol 2012;78:5111-5118.
11.
Wada M, Lkhagvadorj E, Bian L, Wang C, Chiba Y, Nagata S, Shimizu T, Yamashiro Y, Asahara T, Nomoto K: Quantitative reverse transcription-PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus. J Appl Microbiol 2010;108:779-788.
12.
Kamiya S, Nagino M, Kanazawa H, Komatsu S, Mayumi T, Takagi K, Asahara T, Nomoto K, Tanaka R, Nimura Y: The value of bile replacement during external biliary drainage: an analysis of intestinal permeability, integrity, and microflora. Ann Surg 2004;239:510-517.
13.
Hayakawa M, Asahara T, Henzan N, Murakami H, Yamamoto H, Mukai N, Minami Y, Sugano M, Kubota N, Uegaki S, Kamoshida H, Sawamura A, Nomoto K, Gando S: Dramatic changes of the gut flora immediately after severe and sudden insults. Dig Dis Sci 2011;56:2361-2365.
14.
Scholtens PA, Oozeer R, Martin R, Amor KB, Knol J: The early settlers: intestinal microbiology in early life. Annu Rev Food Sci Technol 2012;3:425-447.
15.
Funkhouser LJ, Bordenstein SR: Mom knows best: the universality of maternal microbial transmission. PLoS Biol 2013;11:e1001631.
16.
Gronlund MM, Lehtonen OP, Eerola E, Kero P: Fecal microflora in healthy infants born by different methods of delivery: permanent changes in intestinal flora after cesarean delivery. J Pediatr Gastroenterol Nutr 1999;28:19-25.
17.
Schwiertz A, Gruhl B, Lobnitz M, Michel P, Radke M, Blaut M: Development of the intestinal bacterial composition in hospitalized preterm infants in comparison with breast-fed, full-term infants. Pediatr Res 2003;54:393-399.
18.
Tsuji H, Oozeer R, Matsuda K, Matsuki T, Ohta T, Nomoto K, Tanaka R, Kawashima M, Kawashima K, Nagata S, Yamashiro Y: Molecular monitoring of the development of intestinal microbiota in Japanese infants. Benef Microbes 2012;3:113-125.
19.
Ruemmele FM, Bier D, Marteau P, Rechkemmer G, Bourdet-Sicard R, Walker WA, Goulet O: Clinical evidence for immunomodulatory effects of probiotic bacteria. J Pediatr Gastroenterol Nutr 2009;48:126-141.
20.
Jin HZ, Fan XB, Hang XM, Li KB, Yang H: [Analysis of the probiotic Bifidobacterium and Lactobacillus community in child intestinal flora]. Wei Sheng Wu Xue Bao 2005;45:567-570.
21.
Buccigrossi V, Nicastro E, Guarino A: Functions of intestinal microflora in children. Curr Opin Gastroenterol 2013;29:31-38.
22.
Lin A, Bik EM, Costello EK, Dethlefsen L, Haque R, Relman DA, Singh U: Distinct distal gut microbiome diversity and composition in healthy children from Bangladesh and the United States. PLoS One 2013;8:e53838.
23.
Isolauri E: Development of healthy gut microbiota early in life. J Paediatr Child Health 2012;48(suppl 3):1-6.
24.
Luoto R, Kalliomäki M, Laitinen K, Isolauri E: The impact of perinatal probiotic intervention on the development of overweight and obesity: follow-up study from birth to 10 years. Int J Obes (Lond) 2010;34:1531-1537.
25.
Cutfield WS, Hofman PL, Mitchell M, Morison IM: Could epigenetics play a role in the developmental origins of health and disease? Pediatr Res 2007;61(5 pt 2):68R-75R.
26.
Bezirtzoglou E, Romond C: Occurrence of Enterobacteriaceae and E. coli in the intestine of the newborn by caesarean section. Acta Microbiol Bulg 1990;25:76-81.
27.
Yamashiro Y, Nagata S: Beneficial microbes for premature infants, and children with malignancy undergoing chemotherapy. Benef Microbes 2010;1:357-365.
28.
Bäckhed F: Programming of host metabolism by the gut microbiota. Ann Nutr Metab 2011;58(suppl 2):44-52.
29.
Tojo R, Suárez A, Clemente MG, de los Reyes-Gavilán CG, Margolles A, Gueimonde M, Ruas-Madiedo P: Intestinal microbiota in health and disease: role of bifidobacteria in gut homeostasis. World J Gastroenterol 2014;20:15163-15176.
30.
Lkhagvadorj E, Nagata S, Wada M, Bian L, Wang C, Chiba Y, Yamashiro Y, Shimizu T, Asahara T, Nomoto K: Anti-infectious activity of synbiotics in a novel mouse model of methicillin-resistant Staphylococcus aureus infection. Microbiol Immunol 2010;54:265-275.
31.
Fukuda S, Toh H, Hase K, Oshima K, Nakanishi Y, Yoshimura K, Tobe T, Clarke JM, Topping DL, Suzuki T, Taylor TD, Itoh K, Kikuchi J, Morita H, Hattori M, Ohno H: Bifidobacteria can protect from enteropathogenic infection through production of acetate. Nature 2011;469:543-547.
32.
Rombeau JL, Reilly KJ, Rolandelli RH: Short-chain fatty acids and intestinal surgery: rationale and clinical implication; in Cummings JH, Rombeau JL, Sakata T (eds): Physiology and Clinical Aspects of Short-Chain Fatty Acids. New York, Cambridge University Press, 1995, pp 401-425.
33.
Nagata S, Asahara T, Ohta T, Yamada T, Kondo S, Bian L, Wang C, Yamashiro Y, Nomoto K: Effect of the continuous intake of probiotic-fermented milk containing Lactobacillus casei strain Shirota on fever in a mass outbreak of norovirus gastroenteritis and the faecal microflora in a health service facility for the aged. Br J Nutr 2011;106:549-556.
34.
Bian L, Nagata S, Asahara T, Rahman MS, Ohta T, Yuki N, Wang C, Takano K, Daibo M, Nomoto K, Yamashiro Y: Effects of the continuous intake of Lactobacillus casei strain Shirota-fermented milk on risk management of long term inpatients at health service facilities for the elderly. Int J Probiotics Prebiotics 2011;6:123-132.
35.
Matsumoto K, Takada T, Shimizu K, Kado Y, Kawakami K, Makino I, Yamaoka Y, Hirano K, Nishimura A, Kajimoto O, Nomoto K: The effects of a probiotic milk product containing Lactobacillus casei strain Shirota on the defecation frequency and the intestinal microflora of sub-optimal health state volunteers: a randomized placebo-controlled cross-over study. Biosci Microflora 2006;25:39-48.
36.
Shioiri T, Yahagi K, Nakayama S, Asahara T, Yuki N, Kawakami K, Yamaoka Y, Sakai Y, Nomoto K, Totani M: The effects of a synbiotic milk product containing Lactobacillus casei strain Shirota and transgalactosylated oligosaccharides on defecation frequency, intestinal microflora, organic acid concentrations, and putrefactive metabolite of sub-optimal health state volunteers: a randomized placebo-controlled cross-over study. Biosci Microflora 2006;25:137-146.
37.
Matsumoto K, Takada T, Shimizu K, Moriyama K, Kawakami K, Hirano K, Kajimoto O, Nomoto K: Effects of a probiotic fermented milk beverage containing Lactobacillus casei strain Shirota on defecation frequency, intestinal microbiota, and the intestinal environment of healthy individuals with soft stools. J Biosci Bioeng 2010;110:547-552.
38.
Ostling CE, Lindgren SE: Inhibition of enterobacteria and Listeria growth by lactic, acetic and formic acids. J Appl Bacteriol 1993;75:18-24.
39.
Eklund T: The antimicrobial effect of dissociated and undissociated sorbic acid at different pH levels. J Appl Bacteriol 1983;54:383-389.
40.
Brocklehurst TF, Lund BM: The influence of pH, temperature and organic acids on the initiation of growth of Yersinia enterocolitica. J Appl Bacteriol 1990;69:390-397.
41.
Spanhaak S, Havenaar R, Schaafsma G: The effect of consumption of milk fermented by Lactobacillus casei strain Shirota on the intestinal microflora and immune parameters in humans. Eur J Clin Nutr 1998;52:899-907.
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2015
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