Prebiotic oligosaccharides: Comparative evaluation using in vitro cultures of infants' fecal microbiomes

Applied and Environmental Microbiology

Volumn 80, Issue 23, 2014, Pages 7388-7397

  Stiverson, J ^a   Williams, T ^a,b   Chen, J ^a   Adams, S ^a   Hustead, D ^b   Price, P ^b   Guerrieri, J ^c   Deacon, J ^c   Yu, Z ^a  

^a OHIO STATE UNIVERSITY   (United States)

^b ABBOTT LABORATORIES   (United States)

^c Institute of Clinical Research   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ACETIC ACID; BACTERIA; ESCHERICHIA COLI; FATTY ACIDS; OLIGOSACCHARIDES; ORGANIC ACIDS; PH; POLYMERASE CHAIN REACTION; POLYSACCHARIDES; RNA;

ACETIC ACID CONCENTRATION; BIFIDOBACTERIUM; BIFIDOBACTERIUM LONGUM; COMPARATIVE EVALUATIONS; GALACTO-OLIGOSACCHARIDES; INTESTINAL BIFIDOBACTERIA; LONG-CHAIN INULINS; QUANTITATIVE PCR;

VOLATILE FATTY ACIDS;

ABUNDANCE; ACETIC ACID; BACTERIUM; CARBOHYDRATE; CONCENTRATION (COMPOSITION); FATTY ACID; FECAL COLIFORM; GROWTH RATE; INHIBITION; INOCULATION; POPULATION STRUCTURE;

BACTERIAL DNA; FATTY ACID; OLIGOSACCHARIDE; PREBIOTIC AGENT; RIBOSOME DNA; RNA 16S;

BACTERIAL LOAD; CHEMISTRY; DNA SEQUENCE; DRUG EFFECTS; FECES; GENETICS; HUMAN; IN VITRO STUDY; INFANT; INTESTINE FLORA; METABOLISM; MICROBIOLOGY; MICROFLORA; PH; REAL TIME POLYMERASE CHAIN REACTION;

BACTERIAL LOAD; DNA, BACTERIAL; DNA, RIBOSOMAL; FATTY ACIDS; FECES; GASTROINTESTINAL MICROBIOME; HUMANS; HYDROGEN-ION CONCENTRATION; IN VITRO TECHNIQUES; INFANT; MICROBIOTA; OLIGOSACCHARIDES; PREBIOTICS; REAL-TIME POLYMERASE CHAIN REACTION; RNA, RIBOSOMAL, 16S; SEQUENCE ANALYSIS, DNA;

EID: 84910125468     PISSN: 00992240     EISSN: 10985336     Source Type: Journal    
DOI: 10.1128/AEM.02200-14     Document Type: Article

References (78)

1. Fanaro S, Chierici R, Guenini P, Vigi V. 2003. Intestinal microflora in early infancy: composition and development. Acta Paediatr. 92:S48-S55. http://dx.doi.org/10.1111/j.1651-2227.2003.tb00646.x.
2. Harmsen HJM, Wildeboer-Veloo ACM, Raangs GC, Wagendorp AA, Klijn N, Bindels JG, Welling GW. 2000. Analysis of intestinal flora development in breast-fed and formula-fed infants by using molecular identification and detection methods. J. Pediatr. Gastroenterol. Nutr. 30: 61-67. http://dx.doi.org/10.1097/00005176-200001000-00019.
3. Cebra JJ. 1999. Influences of microbiota on intestinal immune system development. Am. J. Clin. Nutr. 69:1046S-1051S.
4. Chierici R, Fanaro S, Saccomandi D, Vigi V. 2003. Advances in the modulation of the microbial ecology of the gut in early infancy. Acta Paediatr. 92: 56-63. http://dx.doi.org/10.1111/j.1651-2227.2003.tb00647.x.
5. Lönnerdal B. 2004. Human milk proteins: key components for the biological activity of human milk. Adv. Exp. Med. Biol. 554:11-25. http://dx.doi.org/10.1007/978-1-4757-4242-8_4.
6. Horwood LJ, Darlow BA, Mogridge N. 2001. Breast milk feeding and cognitive ability at 7-8 years. Arch. Dis. Child. Fetal Neonatal Ed. 84:F23-F27. http://dx.doi.org/10.1136/fn.84.1.F23.
7. Savino F, Lupica MM. 2006. Breast milk: biological constituents for health and well-being in infancy. Recenti Prog. Med. 97:519-527. (In Italian.) http://dx.doi.org/10.1701/183.1934.
8. Barile D, Rastall RA. 2013. Human milk and related oligosaccharides as prebiotics. Curr. Opin. Biotechnol. 24:214-219. http://dx.doi.org/10.1016/j.copbio.2013.01.008.
9. Coppa GV, Zampini L, Galeazzi T, Gabrielli O. 2006. Prebiotics in human milk: a review. Dig. Liver Dis. 38(Suppl 2):S291-S294. http://dx.doi.org/10.1016/S1590-8658(07)60013-9.
10. Boehm G, Stahl B. 2003. Oligosaccharides, p 203-243. In Mattila-Sandholm T (ed), Functional dairy products. Woodhead Publishing Limited, Cambridge, England.
11. Newburg DS, Ruiz-Palacios GM, Morrow AL. 2005. Human milk glycans protect infants against enteric pathogens. Annu. Rev. Nutr. 25:37-58. http://dx.doi.org/10.1146/annurev.nutr.25.050304.092553.
12. Vandenplas Y. 2002. Oligosaccharides in infant formula. Br. J. Nutr. 87:293-296. http://dx.doi.org/10.1079/BJNBJN/2002551.
13. Roberfroid M. 2007. Prebiotics: the concept revisited. J. Nutr. 137:830S-837S.
14. Boehm G, Moro G. 2008. Structural and functional aspects of prebiotics used in infant nutrition. J. Nutr. 138:1818S-1828S.
15. Williams TA. 2009. A DNA-based investigation of intestinal microbiota of infants and the impact of prebiotics and maternal intestinal microbiota. Ph.D. dissertation. The Ohio State University, Columbus, OH.
16. Ben XM, Zhou XY, Zhao WH, Yu WL, Pan W, Zhang WL, Wu SM, Van Beusekom CM, Schaafsma A. 2004. Supplementation of milk formula with galacto-oligosaccharides improves intestinal micro-flora and fermentation in term infants. Chin. Med. J. (Engl.) 117:927-931.
17. Boehm G, Lidestri M, Casetta P, Jelinek J, Negretti F, Stahl B, Marini A. 2002. Supplementation of a bovine milk formula with an oligosaccharide mixture increases counts of faecal bifidobacteria in preterm infants. Arch. Dis. Child. Fetal Neonatal Ed. 86:F178-F181. http://dx.doi.org/10.1136/fn.86.3.F178.
18. Kim SH, Lee DH, Meyer D. 2007. Supplementation of baby formula with native inulin has a prebiotic effect in formula-fed babies. Asia Pac. J. Clin. Nutr. 16:172-177.
19. Brunser O, Gotteland M, Cruchet S, Figueroa G, Garrido D, Steenhout P. 2006. Effect of a milk formula with prebiotics on the intestinal microbiota of infants after an antibiotic treatment. Pediatr. Res. 59:451-456. http://dx.doi.org/10.1203/01.pdr.0000198773.40937.61.
20. Euler AR, Mitchell DK, Kline R, Pickering LK. 2005. Prebiotic effect of fructo-oligosaccharide supplemented term infant formula at two concentrations compared with unsupplemented formula and human milk. J. Pediatr. Gastroenterol. Nutr. 40:157-164. http://dx.doi.org/10.1097/00005176-200502000-00014.
21. Nakamura N, Gaskins HR, Collier CT, Nava GM, Rai D, Petschow B, Russell WM, Harris C, Mackie RI, Wampler JL, Walker DC. 2009. Molecular ecological analysis of fecal bacterial populations from term infants fed formula supplemented with selected blends of prebiotics. Appl. Environ. Microbiol. 75:1121-1128. http://dx.doi.org/10.1128/AEM.02359-07.
22. Kurokawa K, Itoh T, Kuwahara T, Oshima K, Toh H, Toyoda A, Takami H, Morita H, Sharma VK, Srivastava TP, Taylor TD, Noguchi H, Mori H, Ogura Y, Ehrlich DS, Itoh K, Takagi T, Sakaki Y, Hayashi T, Hattori M. 2007. Comparative metagenomics revealed commonly enriched gene sets in human gut microbiomes. DNA Res. 14:169-181. http://dx.doi.org/10.1093/dnares/dsm018.
23. Magne F, Hachelaf W, Suau A, Boudraa G, Mangin I, Touhami M, Bouziane-Nedjadi K, Pochart P. 2006. A longitudinal study of infant faecal microbiota during weaning. FEMS Microbiol. Ecol. 58:563-571. http://dx.doi.org/10.1111/j.1574-6941.2006.00182.x.
24. Palmer C, Bik EM, Digiulio DB, Relman DA, Brown PO. 2007. Development of the human infant intestinal microbiota. PLoS Biol. 5:e177. http://dx.doi.org/10.1371/journal.pbio.0050177.
25. Amaretti A, Bernardi T, Tamburini E, Zanoni S, Lomma M, Matteuzzi D, Rossi M. 2007. Kinetics and metabolism of Bifidobacterium adolescentis MB 239 growing on glucose, galactose, lactose, and galactooligosaccharides. Appl. Environ. Microbiol. 73:3637-3644. http://dx.doi.org/10.1128/AEM.02914-06.
26. Rada V, Nevoral J, Trojanová I, Tománková E, Smehilová M, Killer J. 2008. Growth of infant faecal bifidobacteria and clostridia on prebiotic oligosaccharides in in vitro conditions. Anaerobe 14:205-208. http://dx.doi.org/10.1016/j.anaerobe.2008.05.003.
27. Vernazza CL, Gibson GR, Rastall RA. 2006. Carbohydrate preference, acid tolerance and bile tolerance in five strains of Bifidobacterium. J. Appl. Microbiol. 100:846-853. http://dx.doi.org/10.1111/j.1365-2672.2006.02832.x.
28. Olano-Martin E, Gibson GR, Rastell RA. 2002. Comparison of the in vitro bifidogenic properties of pectins and pectic-oligosaccharides. J. Appl. Microbiol. 93:505-511. http://dx.doi.org/10.1046/j.1365-2672.2002.01719.x.
29. Olano-Martin E, Mountzouris KC, Gibson GR, Rastall RA. 2000. In vitro fermentability of dextran, oligodextran and maltodextrin by human gut bacteria. Br. J. Nutr. 83:247-255.
30. Rycroft CE, Jones MR, Gibson GR, Rastall RA. 2001. A comparative in vitro evaluation of the fermentation properties of prebiotic oligosaccharides. J. Appl. Microbiol. 91:878-887. http://dx.doi.org/10.1046/j.1365-2672.2001.01446.x.
31. Yu Z, Morrison M. 2009. The gut microbiome: current understanding and future perspective, p 19-40. In Jaykus L-A, Wang H, Schlesinger LS (ed), Food-borne microbes: shaping the host ecosystem. ASM Press, Washington, DC.
32. Veereman-Wauters G, Staelens S, Van de Broek H, Plaskie K, Wesling F, Roger L, McCartney A, Assam P. 2011. Physiological and bifidogenic effects of prebiotic supplements in infant formulae. J. Pediatr. Gastroenterol. Nutr. 52:763-771. http://dx.doi.org/10.1097/MPG.0b013e3182139f39.
33. Moro G, Minoli I, Mosca M, Fanaro S, Jelinek J, Stahl B, Boehm G. 2002. Dosage-related bifidogenic effects of galacto-and fructo-oligosaccharides in formula-fed term infants. J. Pediatr. Gastroenterol. Nutr. 34:291-295. http://dx.doi.org/10.1097/00005176-200203000-00014.
34. Sanz ML, Gibson GR, Rastall RA. 2005. Influence of disaccharide structure on prebiotic selectivity in vitro. J. Agric. Food Chem. 53:5192-5199. http://dx.doi.org/10.1021/jf050276w.
35. Knol J, Scholtens P, Kafka C, Steenbakkers J, Gro S, Helm K, Klarczyk M, Schöpfer H, Böckler H-M, Wells J. 2005. Colon microflora in infants fed formula with galacto-and fructo-oligosaccharides: more like breastfed infants. J. Pediatr. Gastroenterol. Nutr. 40:36-42. http://dx.doi.org/10.1097/00005176-200501000-00007.
36. Shuhaimi M, Ali AM, Saleh NM, Yazid AM. 2001. Utilisation of enterobacterial repetitive intergenic consensus (ERIC) sequence-based PCR to fingerprint the genomes of Bifidobacterium isolates and other probiotic bacteria. Biotechnol. Lett. 23:731-736. http://dx.doi.org/10.1023/A:1010355003674.
37. Ventura M, Meylan V, Zink R. 2003. Identification and tracing of Bifidobacterium species by use of enterobacterial repetitive intergenic consensus sequences. Appl. Environ. Microbiol. 69:4296-4301. http://dx.doi.org/10.1128/AEM.69.7.4296-4301.2003.
38. Lane DJ. 1991. 16S/23S rRNA sequencing, p 115-175. In Stackebrandt E, Goodfellow MD (ed), Nucleic acid techniques in bacterial systematics. John Wiley and Sons, New York, NY.
39. Yu Z, Morrison M. 2004. Improved extraction of PCR-quality community DNA from digesta and fecal samples. Biotechniques 36:808-812.
40. Anderson K, Yu Z, Chen J, Jenkins J, Courtney P, Morrison M. 2008. Analyses of Bifidobacterium, Lactobacillus, and total bacterial populations in healthy volunteers consuming calcium gluconate by denaturing gradient gel electrophoresis and real-time PCR. Int. J. Probiotics Prebiotics 3:31-36.
41. Bartosch S, Fite A, Macfarlane GT, McMurdo MET. 2004. Characterization of bacterial communities in feces from healthy elderly volunteers and hospitalized elderly patients by using real-time PCR and effects of antibiotic treatment on the fecal microbiota. Appl. Environ. Microbiol. 70:3575-3581. http://dx.doi.org/10.1128/AEM.70.6.3575-3581.2004.
42. Matsuki T, Watanabe K, Fujimoto J, Kado Y, Takada T, Matsumoto K, Tanaka R. 2004. Quantitative PCR with 16S rRNA-gene-targeted speciesspecific primers for analysis of human intestinal bifidobacteria. Appl. Environ. Microbiol. 70:167-173. http://dx.doi.org/10.1128/AEM.70.1.167-173.2004.
43. Xia Q, Williams T, Hustead D, Price P, Morrison M, Yu Z. 2012. Quantitative analysis of intestinal bacterial populations from term infants fed formula supplemented with fructo-oligosaccharides. J. Pediatr. Gastroenterol. Nutr. 55(3):314-320. http://dx.doi.org/10.1097/MPG.0b013e3182523254.
44. Sabat G, Rose P, Hickey WJ, Harkin JM. 2000. Selective and sensitive method for PCR amplification of Escherichia coli 16S rRNA genes in soil. Appl. Environ. Microbiol. 66:844-849. http://dx.doi.org/10.1128/AEM.66.2.844-849.2000.
45. Chen J, Yu Z, Michel FC, Jr, Wittum T, Morrison M. 2007. Development and application of real-time PCR assays for quantification of erm genes conferring resistance to macrolides-lincosamides-streptogramin B in livestock manure and manure management systems. Appl. Environ. Microbiol. 73:4407-4416. http://dx.doi.org/10.1128/AEM.02799-06.
46. Marchesi JR, Sato T, Weightman AJ, Martin TA, Fry JC, Hiom SJ, Wade WG. 1998. Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA. Appl. Environ. Microbiol. 64:795-799.
47. Osborn AM, Moore ER, Timmis KN. 2000. An evaluation of terminalrestriction fragment length polymorphism (T-RFLP) analysis for the study of microbial community structure and dynamics. Environ. Microbiol. 2:39-50. http://dx.doi.org/10.1046/j.1462-2920.2000.00081.x.
48. Yu Z, Garcia-Gonzalez R, Schanbacher FL, Morrison M. 2008. Evaluations of different hypervariable regions of archaeal 16S rRNA genes in profiling of methanogens by archaea-specific PCR and denaturing gradient gel electrophoresis. Appl. Environ. Microbiol. 74:889-893. http://dx.doi.org/10.1128/AEM.00684-07.
49. Marteau P, Seksik P. 2004. Tolerance of probiotics and prebiotics. J. Clin. Gastroenterol. 38:S67-S69. http://dx.doi.org/10.1097/01.mcg.0000128929.37156.a7.
50. Mugambi MN, Musekiwa A, Lombard M, Young T, Blaauw R. 2012. Synbiotics, probiotics or prebiotics in infant formula for full term infants: a systematic review. Nutr. J. 11:81. http://dx.doi.org/10.1186/1475-2891-11-81.
51. Rao S, Srinivasjois R, Patole S. 2009. Prebiotic supplementation in full-term neonates: a systematic review of randomized controlled trials. Arch. Pediatr. Adolesc. Med. 163:755-764. http://dx.doi.org/10.1001/archpediatrics.2009.94.
52. Roberfroid M, Gibson GR, Hoyles L, McCartney AL, Rastall R, Rowland I, Wolvers D, Watzl B, Szajewska H, Stahl B, Guarner F, Respondek F, Whelan K, Coxam V, Davicco MJ, Leotoing L, Wittrant Y, Delzenne NM, Cani PD, Neyrinck AM, Meheust A. 2010. Prebiotic effects: metabolic and health benefits. Br. J. Nutr. 104(Suppl 2):S1-S63. http://dx.doi.org/10.1017/S0007114510003363.
53. Macfarlane GT, Steed H, Macfarlane S. 2008. Bacterial metabolism and health-related effects of galacto-oligosaccharides and other prebiotics. J. Appl. Microbiol. 104:305-344. http://dx.doi.org/10.1111/j.1365-2672.2007.03520.x.
54. Zivkovic AM, German JB, Lebrilla CB, Mills DA. 2 August 2010. Human milk glycobiome and its impact on the infant gastrointestinal microbiota. Proc. Natl. Acad. Sci. U. S. A. http://dx.doi.org/10.1073/pnas.1000083107.
55. Garrido D, Ruiz-Moyano S, Jimenez-Espinoza R, Eom HJ, Block DE, Mills DA. 2013. Utilization of galactooligosaccharides by Bifidobacterium longum subsp. infantis isolates. Food Microbiol. 33:262-270. http://dx.doi.org/10.1016/j.fm.2012.10.003.
56. Pompei A, Cordisco L, Raimondi S, Amaretti A, Pagnoni UM, Matteuzzi D, Rossi M. 2008. In vitro comparison of the prebiotic effects of two inulin-type fructans. Anaerobe 14:280-286. http://dx.doi.org/10.1016/j.anaerobe.2008.07.002.
57. Rossi M, Corradini C, Amaretti A, Nicolini M, Pompei A, Zanoni S, Matteuzzi D. 2005. Fermentation of fructooligosaccharides and inulin by bifidobacteria: a comparative study of pure and fecal cultures. Appl. Environ. Microbiol. 71:6150-6158. http://dx.doi.org/10.1128/AEM.71.10.6150-6158.2005.
58. Kleessen B, Hartmann L, Blaut M. 2001. Oligofructose and long-chain inulin: influence on the gut microbial ecology of rats associated with a human faecal flora. Br. J. Nutr. 86:291-300. http://dx.doi.org/10.1079/BJN2001403.
59. Maathuis AJ, van den Heuvel EG, Schoterman MH, Venema K. 2012. Galacto-oligosaccharides have prebiotic activity in a dynamic in vitro colon model using a 13C-labeling technique. J. Nutr. 142:1205-1212. http://dx.doi.org/10.3945/jn.111.157420.
60. Kleessen B, Bunke H, Tovar K, Noack J, Sawatzki G. 1995. Influence of two infant formulas and human milk on the development of faecal flora in newborn infants. Acta Paediatr. 84:1347-1356. http://dx.doi.org/10.1111/j.1651-2227.1995.tb13567.x.
61. Sakata S, Tonooka T, Ishizeki S, Takada M, Sakamoto M, Fukuyama M, Benno Y. 2005. Culture-independent analysis of fecal microbiota in infants, with special reference to Bifidobacterium species. FEMS Microbiol. Lett. 243:417-423. http://dx.doi.org/10.1016/j.femsle.2005.01.002.
62. Avershina E, Storro O, Oien T, Johnsen R, Wilson R, Egeland T, Rudi K. 2013. Bifidobacterial succession and correlation networks in a large unselected cohort of mothers and their children. Appl. Environ. Microbiol. 79:497-507. http://dx.doi.org/10.1128/AEM.02359-12.
63. Asakuma S, Hatakeyama E, Urashima T, Yoshida E, Katayama T, Yamamoto K, Kumagai H, Ashida H, Hirose J, Kitaoka M. 2011. Physiology of consumption of human milk oligosaccharides by infant gut-associated bifidobacteria. J. Biol. Chem. 286:34583-34592. http://dx.doi.org/10.1074/jbc.M111.248138.
64. Garrido D, Dallas DC, Mills DA. 2013. Consumption of human milk glycoconjugates by infant-associated bifidobacteria: mechanisms and implications. Microbiology 159:649-664. http://dx.doi.org/10.1099/mic.0.064113-0.
65. Ruiz-Moyano S, Totten SM, Garrido D, Smilowitz JT, German JB, Lebrilla CB, Mills DA. 2013. Variation in consumption of human milk oligosaccharides by infant-gut associated strains of Bifidobacterium breve. Appl. Environ. Microbiol. 79:6040-6049. http://dx.doi.org/10.1128/AEM.01843-13.
66. Roger LC, Costabile A, Holland DT, Hoyles L, McCartney AL. 2010. Examination of faecal Bifidobacterium populations in breast-and formula-fed infants during the first 18 months of life. Microbiology 156:3329-3341. http://dx.doi.org/10.1099/mic.0.043224-0.
67. Rinne MM, Gueimonde M, Kalliomäki M, Hoppu U, Salminen SJ, Isolauri E. 2005. Similar bifidogenic effects of prebiotic-supplemented partially hydrolyzed infant formula and breastfeeding on infant gut microbiota. FEMS Immunol. Med. Microbiol. 43:59-65. http://dx.doi.org/10.1016/j.femsim.2004.07.005.
68. Vlkova E, Rada V, Bujnakova D, Kmet V. 2004. Enumeration, isolation, and identification of bifidobacteria from infant feces. Folia Microbiol. (Praha) 49:209-212. http://dx.doi.org/10.1007/BF02931404.
69. Harmsen HJ, Gibson GR, Elfferich P, Raangs GC, Wildeboer-Veloo AC, Argaiz A, Roberfroid MB, Welling GW. 2000. Comparison of viable cell counts and fluorescence in situ hybridization using specific rRNA-based probes for the quantification of human fecal bacteria. FEMS Microbiol. Lett. 183:125-129. http://dx.doi.org/10.1111/j.1574-6968.2000.tb08945.x.
70. Fushinobu S. 2010. Unique sugar metabolic pathways of bifidobacteria. Biosci. Biotechnol. Biochem. 74:2374-2384. http://dx.doi.org/10.1271/bbb.100494.
71. Gomes AMP, Malcata FX. 1999. Bifidobacterium spp. and Lactobacillus acidophilus: biological, biochemical, technological and therapeutical properties relevant for use as probiotics. Trends Food Sci. Technol. 10: 139-157. http://dx.doi.org/10.1016/S0924-2244(99)00033-3.
72. Wilson M. 2005. The gastrointestinal tract and its indigenous microbiota, p 251-317. In Wilson M (ed), Microbial inhabitants of humans. Cambridge University Press, Cambridge, United Kingdom.
73. Ogawa K, Ben RA, Pons S, de Paolo MI, Bustos Fernandez L. 1992. Volatile fatty acids, lactic acid, and pH in the stools of breast-fed and bottle-fed infants. J. Pediatr. Gastroenterol. Nutr. 15:248-252. http://dx.doi.org/10.1097/00005176-199210000-00004.
74. Penders J, Vink C, Driessen C, London N, Thijs C, Stobberingh EE. 2005. Quantification of Bifidobacterium spp., Escherichia coli and Clostridium difficile in faecal samples of breast-fed and formula-fed infants by real-time PCR. FEMS Microbiol. Lett. 243:141-147. http://dx.doi.org/10.1016/j.femsle.2004.11.052.
75. Scott KP, Martin JC, Duncan SH, Flint HJ. 28 August 2013. Prebiotic stimulation of human colonic butyrate-producing bacteria and bifidobacteria, in vitro. FEMS Microbiol. Ecol. http://dx.doi.org/10.1111/1574-6941.12186.
76. Nadkarni MA, Martin FE, Jacques NA, Hunter N. 2002. Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set. Microbiology 148:257-266.
77. Bernhard AE, Field KG. 2000. A PCR assay to discriminate human and ruminant feces on the basis of host differences in Bacteroides-Prevotella genes encoding 16S rRNA. Appl. Environ. Microbiol. 66:4571-4574. http://dx.doi.org/10.1128/AEM.66.10.4571-4574.2000.
78. Satokari RM, Vaughan EE, Akkermans ADL, Saarela M, de Vos WM. 2001. Bifidobacterial diversity in human feces detected by genus-specific PCR and denaturing gradient gel electrophoresis. Appl. Environ. Microbiol. 67:504-513. http://dx.doi.org/10.1128/AEM.67.2.504-513.2001.