Effect of a multi-species synbiotic formulation on fecal bacterial microbiota of healthy cats and dogs as evaluated by pyrosequencing
Jose F. Garcia-Mazcorro
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorDavid J. Lanerie
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorCasey G. Paddock
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorNiels Grützner
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorJörg M. Steiner
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorRenata Ivanek
Department of Veterinary Integrative Biosciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorCorresponding Author
Jan S. Suchodolski
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Correspondence: Jan S. Suchodolski, Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843-4474, USA. Tel./fax: +1 979 458 4015; e-mail: [email protected]Search for more papers by this authorJose F. Garcia-Mazcorro
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorDavid J. Lanerie
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorCasey G. Paddock
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorNiels Grützner
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorJörg M. Steiner
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorRenata Ivanek
Department of Veterinary Integrative Biosciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Search for more papers by this authorCorresponding Author
Jan S. Suchodolski
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA
Correspondence: Jan S. Suchodolski, Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843-4474, USA. Tel./fax: +1 979 458 4015; e-mail: [email protected]Search for more papers by this authorAbstract
The effect of a multi-species synbiotic on the fecal microbiota of healthy cats (n = 12) and dogs (n = 12) was evaluated. The synbiotic (containing 5 × 109 CFU of a mixture of seven probiotic strains, and a blend of fructooligosaccharides and arabinogalactans) was administered daily for 21 days. Fecal and serum samples were collected before, during, and up to 3 weeks after administration. Changes in the fecal microbiota were analyzed using denaturing gradient gel electrophoresis, 16S rRNA gene libraries, quantitative real-time PCR, and 16S rRNA gene 454-pyrosequencing. Probiotic species were detectable in 10/12 dogs and 11/12 cats during product administration. Abundances of Enterococcus and Streptococcus spp. were significantly increased in at least one time point during administration, and returned to baseline abundance after treatment was discontinued. No changes in the major bacterial phyla were identified on 454-pyrosequencing. No adverse gastrointestinal effects were recorded and no significant changes in gastrointestinal function or immune markers were observed during the study period. This study shows that while the ingestion of probiotics and prebiotics does not appear to alter the predominant bacterial phyla present in feces, supplementation with the investigated synbiotic leads to an increased abundance of probiotic bacteria in the feces of healthy cats and dogs.
Supporting Information
| Filename | Description |
|---|---|
| fem1185-sup-0001-TableS1.docWord document, 94 KB | Table S1. Median (interquartile range) pyrosequencing bacterial tags (percentages of all sequences) obtained before synbiotic administration at baseline (BL), after 5 days of administration of the synbiotic formulation (day 5), and 2 days after discontinuation of administration of the synbiotic formulation (day 23) in the enrolled cats. |
| fem1185-sup-0002-TableS2.docWord document, 68.5 KB | Table S2. Median (interquartile range) pyrosequencing bacterial tags (percentages of all sequences) obtained before synbiotic administration at baseline (BL), after 5 days of administration of the synbiotic formulation (day 5), and 2 days after discontinuation of administration of the synbiotic formulation (day 23) in the enrolled dogs. |
| fem1185-sup-0003-TableS3.docWord document, 60 KB | Table S3. Mean concentrations (± SD) of relevant blood, serum, and fecal markers obtained before (baseline), during (day 21), and after (day 42) synbiotic administration. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
References
- Baillon MLA, Marshall-Jones ZV & Butterwick RF (2004) Effects of probiotic Lactobacillus acidophilus strain DSM13241 in healthy adult dogs. Am J Vet Res 65: 338–343.
- Barry KA, Hernot DC, Middelbos IS, Francis C, Dunsford B, Swanson KS & Fahey GC Jr. (2009) Low-level fructan supplementation of dogs enhances nutrient digestion and modifies stool metabolite concentrations, but does not alter fecal microbiota populations. J Anim Sci 87: 3244–3252.
- Bartosch S, Woodmansey EJ, Paterson JC, McMurdo ME & Macfarlane GT (2005) Microbiological effects of consuming a synbiotic containing Bifidobacterium bifidum, Bifidobacterium lactis, and oligofructose in elderly persons, determined by real-time polymerase chain reaction and counting of viable bacteria. Clin Infect Dis 40: 28–37.
- Benjamini Y & Hochberg Y (1995) Controlling the false discovery rate – a practical and powerful approach to multiple testing. J R Stat Soc Ser B Methodol 57: 289–300.
- Benyacoub J, Czarnecki-Maulden GL, Cavadini C, Sauthier T, Anderson RE, Schiffrin EJ & von der Weid T (2003) Supplementation of food with Enterococcus faecium (SF68) stimulates immune functions in young dogs. J Nutr 133: 1158–1162.
- Bezkorovainy A (2001) Probiotics: determinants of survival and growth in the gut. Am J Clin Nutr 73: 399S–405S.
- Biagi G, Cipollini I, Pompei A, Zaghini G & Matteuzzi D (2007) Effect of a Lactobacillus animalis strain on composition and metabolism of the intestinal microflora in adult dogs. Vet Microbiol 124: 160–165.
- Blount DG, Pritchard DI & Heaton PR (2005) Age-related alterations to immune parameters in Labrador retriever dogs. Vet Immunol Immunopathol 108: 399–407.
- Callaway TR, Edrington TS, Anderson RC, Harvey RB, Genovese KJ, Kennedy CN, Venn DW & Nisbet DJ (2008) Probiotics, prebiotics and competitive exclusion for prophylaxis against bacterial disease. Anim Health Res Rev 9: 217–225.
- Campbell DJ, Rawlings JM, Koelsch S, Wallace J, Strain JJ & Hannigan BM (2004) Age-related differences in parameters of feline immune status. Vet Immunol Immunopathol 100: 73–80.
- Culligan EP, Hill C & Sleator RD (2009) Probiotics and gastrointestinal disease: successes, problems and future prospects. Gut Pathog 1: 19.
- Eckburg PB, Bik EM, Bernstein CN, Purdom E, Dethlefsen L, Sargent M, Gill SR, Nelson KE & Relman DA (2005) Diversity of the human intestinal microbial flora. Science 308: 1635–1638.
- Elli M, Callegari ML, Ferrari S, Bessi E, Cattivelli D, Soldi S, Morelli L, Goupil Feuillerat N & Antoine JM (2006) Survival of yogurt bacteria in the human gut. Appl Environ Microbiol 72: 5113–5117.
- FAO/WHO (2002) Working Group Report on Drafting Guidelines for the Evaluation of Probiotics in Food. FAO/WHO, London, ON.
- Fetz K, Ruaux CG, Steiner JM, Suchodolski JS & Williams DA (2004) Purification and partial characterization of feline α1-proteinase inhibitor (fα1-PI) and the development and validation of a radioimmunoassay for the measurement of fα1-PI in serum. Biochimie 86: 67–75.
- Frank DN, St Amand AL, Feldman RA, Boedeker EC, Harpaz N & Pace NR (2007) Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases. P Natl Acad Sci USA 104: 13780–13785.
- Fuller R (1989) Probiotics in man and animals. J Appl Bacteriol 66: 365–378.
- Furet JP, Quenee P & Tailliez P (2004) Molecular quantification of lactic acid bacteria in fermented milk products using real-time quantitative PCR. Int J Food Microbiol 97: 197–207.
- Furrie E (2006) A molecular revolution in the study of intestinal microflora. Gut 55: 141–143.
- Gibson GR, Scott KP, Rastall RA et al. (2010) Dietary prebiotics: current status and new definition. Food Sci Technol Bull Funct Foods 7: 1–19.
- Handl S, Dowd SE, Garcia-Mazcorro JF, Steiner JM & Suchodolski JS (2011) Massive parallel 16S rRNA gene pyrosequencing reveals highly diverse fecal bacterial and fungal communities in healthy dogs and cats. FEMS Microbiol Ecol 76: 301–310.
- Heilig HG, Zoetendal EG, Vaughan EE, Marteau P, Akkermans AD & de Vos WM (2002) Molecular diversity of Lactobacillus spp. and other lactic acid bacteria in the human intestine as determined by specific amplification of 16S ribosomal DNA. Appl Environ Microbiol 68: 114–123.
- Jones SE & Versalovic J (2009) Probiotic Lactobacillus reuteri biofilms produce antimicrobial and anti-inflammatory factors. BMC Microbiol 9: 35.
- Kailasapathy K & Chin J (2000) Survival and therapeutic potential of probiotic organisms with reference to Lactobacillus acidophilus and Bifidobacterium spp. Immunol Cell Biol 78: 80–88.
- Kelley RL, Minikhiem D, Kiely B, O'Mahony L, O'Sullivan D, Boileau T & Park JS (2009) Clinical benefits of probiotic canine-derived Bifidobacterium animalis strain AHC7 in dogs with acute idiopathic diarrhea. Vet Ther 10: 121–130.
- Lappin MR, Veir JK, Satyaraj E & Czarnecki-Maulden G (2009) Pilot study to evaluate the effect of oral supplementation of Enterococcus faecium SF68 on cats with latent feline herpesvirus 1. J Feline Med Surg 11: 650–654.
- Lee YK, Puong KY, Ouwehand AC & Salminen S (2003) Displacement of bacterial pathogens from mucus and Caco-2 cell surface by lactobacilli. J Med Microbiol 52: 925–930.
- Lozupone C & Knight R (2005) UniFrac: a new phylogenetic method for comparing microbial communities. Appl Environ Microbiol 71: 8228–8235.
- Malinen E, Rinttila T, Kajander K, Matto J, Kassinen A, Krogius L, Saarela M, Korpela R & Palva A (2005) Analysis of the fecal microbiota of irritable bowel syndrome patients and healthy controls with real-time PCR. Am J Gastroenterol 100: 373–382.
- Marshall-Jones ZV, Baillon ML, Croft JM & Butterwick RF (2006) Effects of Lactobacillus acidophilus DSM13241 as a probiotic in healthy adult cats. Am J Vet Res 67: 1005–1012.
- Martin R, Olivares M, Perez M, Xaus J, Torre C, Fernandez L & Rodriguez JM (2010) Identification and evaluation of the probiotic potential of lactobacilli isolated from canine milk. Vet J 185: 193–198.
- McCoy S & Gilliland SE (2007) Isolation and characterization of Lactobacillus species having potential for use as probiotic cultures for dogs. J Food Sci 72: M94–M97.
- Melgarejo T, Williams DA & Asem EK (1998) Enzyme-linked immunosorbent assay for canine alpha 1-protease inhibitor. Am J Vet Res 59: 127–130.
- Middelbos IS, Vester Boler BM, Qu A, White BA, Swanson KS & Fahey GC Jr. (2010) Phylogenetic characterization of fecal microbial communities of dogs fed diets with or without supplemental dietary fiber using 454 pyrosequencing. PLoS ONE 5: e9768.
- Muyzer G, de Waal EC & Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59: 695–700.
- Ogue-Bon E, Khoo C, McCartney AL, Gibson GR & Rastall RA (2010) In vitro effects of synbiotic fermentation on the canine faecal microbiota. FEMS Microbiol Ecol 73: 587–600.
- O'Mahony D, Murphy KB, MacSharry J, Boileau T, Sunvold G, Reinhart G, Kiely B, Shanahan F & O'Mahony L (2009) Portrait of a canine probiotic Bifidobacterium – from gut to gut. Vet Microbiol 139: 106–112.
- Pagnini C, Saeed R, Bamias G, Arseneau KO, Pizarro TT & Cominelli F (2010) Probiotics promote gut health through stimulation of epithelial innate immunity. P Natl Acad Sci USA 107: 454–459.
- Palka-Santini M, Schwarz-Herzke B, Hosel M, Renz D, Auerochs S, Brondke H & Doerfler W (2003) The gastrointestinal tract as the portal of entry for foreign macromolecules: fate of DNA and proteins. Mol Genet Genomics 270: 201–215.
- R Development Core Team (2004) A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria. http://www.R-project.org/.
- Rinkinen M, Westermarck E, Salminen S & Ouwehand AC (2003) Absence of host specificity for in vitro adhesion of probiotic lactic acid bacteria to intestinal mucus. Vet Microbiol 97: 55–61.
- Rinttila T, Kassinen A, Malinen E, Krogius L & Palva A (2004) Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR. J Appl Microbiol 97: 1166–1177.
- Ritchie LE, Burke KF, Garcia-Mazcorro JF, Steiner JM & Suchodolski JS (2010) Characterization of fecal microbiota in cats using universal 16S rRNA gene and group-specific primers for Lactobacillus and Bifidobacterium spp. Vet Microbiol 144: 140–146.
- Ritz K (2007) The plate debate: cultivable communities have no utility in contemporary environmental microbial ecology. FEMS Microbiol Ecol 60: 358–362.
10.1111/j.1574-6941.2007.00331.x Google Scholar
- Roberfroid M, Gibson GR & Hoyles L et al. (2010) Prebiotic effects: metabolic and health benefits. Br J Nutr 104 (suppl 2): S1–S63.
- Saulnier DM, Gibson GR & Kolida S (2008) In vitro effects of selected synbiotics on the human faecal microbiota composition. FEMS Microbiol Ecol 66: 516–527.
- Sauter SN, Benyacoub J, Allenspach K, Gaschen F, Ontsouka E, Reuteler G, Cavadini C, Knorr R & Blum JW (2006) Effects of probiotic bacteria in dogs with food responsive diarrhoea treated with an elimination diet. J Anim Physiol Anim Nutr (Berl) 90: 269–277.
- Simpson KW, Rishniw M, Bellosa M, Liotta J, Lucio A, Baumgart M, Czarnecki-Maulden G, Benyacoub J & Bowman D (2009) Influence of Enterococcus faecium SF68 probiotic on giardiasis in dogs. J Vet Intern Med 23: 476–481.
- Sparkes AH, Papasouliotis K, Sunvold G, Werrett G, Clarke C, Jones M, Gruffydd-Jones TJ & Reinhart G (1998) Bacterial flora in the duodenum of healthy cats, and effect of dietary supplementation with fructo-oligosaccharides. Am J Vet Res 59: 431–435.
- Strompfova V, Laukova A & Ouwehand AC (2004) Selection of enterococci for potential canine probiotic additives. Vet Microbiol 100: 107–114.
- Suchodolski JS, Camacho J & Steiner JM (2008) Analysis of bacterial diversity in the canine duodenum, jejunum, ileum, and colon by comparative 16S rRNA gene analysis. FEMS Microbiol Ecol 66: 567–578.
- Suchodolski JS, Ruaux CG, Steiner JM, Fetz K & Williams DA (2004) Application of molecular fingerprinting for qualitative assessment of small-intestinal bacterial diversity in dogs. J Clin Microbiol 42: 4702–4708.
- Suchodolski JS, Ruaux CG, Steiner JM, Fetz K & Williams DA (2005) Assessment of the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique. Am J Vet Res 66: 1556–1562.
- Suchodolski JS, Dowd SE, Westermarck E, Steiner JM, Wolcott RD, Spillmann T & Harmoinen JA (2009) The effect of the macrolide antibiotic tylosin on microbial diversity in the canine small intestine as demonstrated by massive parallel 16S rRNA gene sequencing. BMC Microbiol 9: 210.
- Swanson KS, Grieshop CM, Flickinger EA, Bauer LL, Chow J, Wolf BW, Garleb KA & Fahey GC Jr. (2002) Fructooligosaccharides and Lactobacillus acidophilus modify gut microbial populations, total tract nutrient digestibilities and fecal protein catabolite concentrations in healthy adult dogs. J Nutr 132: 3721–3731.
- Swanson KS, Dowd SE, Suchodolski JS et al. (2011) Phylogenetic and gene-centric metagenomics of the canine intestinal microbiome reveals similarities with humans and mice. ISME J 5: 639–649.
- Tress U, Suchodolski JS, Williams DA & Steiner JM (2006) Development of a fecal sample collection strategy for extraction and quantification of fecal immunoglobulin A in dogs. Am J Vet Res 67: 1756–1759.
- Vahjen W & Manner K (2003) The effect of a probiotic Enterococcus faecium product in diets of healthy dogs on bacteriological counts of Salmonella spp., Campylobacter spp. and Clostridium spp. in faeces. Arch Tierernahr 57: 229–233.
- Valeur N, Engel P, Carbajal N, Connolly E & Ladefoged K (2004) Colonization and immunomodulation by Lactobacillus reuteri ATCC 55730 in the human gastrointestinal tract. Appl Environ Microbiol 70: 1176–1181.
- Venturi A, Gionchetti P, Rizzello F, Johansson R, Zucconi E, Brigidi P, Matteuzzi D & Campieri M (1999) Impact on the composition of the faecal flora by a new probiotic preparation: preliminary data on maintenance treatment of patients with ulcerative colitis. Aliment Pharmacol Ther 13: 1103–1108.
- Vitali B, Ndagijimana M, Cruciani F, Carnevali P, Candela M, Guerzoni ME & Brigidi P (2010) Impact of a synbiotic food on the gut microbial ecology and metabolic profiles. BMC Microbiol 10: 4.
- Walter J, Hertel C, Tannock GW, Lis CM, Munro K & Hammes WP (2001) Detection of Lactobacillus, Pediococcus, Leuconostoc, and Weissella species in human feces by using group-specific PCR primers and denaturing gradient gel electrophoresis. Appl Environ Microbiol 67: 2578–2585.
- Warnes GR (2010) Various R programming tools for plotting data. http://cran.r-project.org/web/packages/gplots/gplots.pdf.
- Wolvers D, Antoine JM, Myllyluoma E, Schrezenmeir J, Szajewska H & Rijkers GT (2010) Guidance for substantiating the evidence for beneficial effects of probiotics: prevention and management of infections by probiotics. J Nutr 140: 698S–712S.
- Worthley DL, Le Leu RK, Whitehall VL et al. (2009) A human, double-blind, placebo-controlled, crossover trial of prebiotic, probiotic, and synbiotic supplementation: effects on luminal, inflammatory, epigenetic, and epithelial biomarkers of colorectal cancer. Am J Clin Nutr 90: 578–586.
- Zoetendal EG, Rajilic-Stojanovic M & de Vos WM (2008) High-throughput diversity and functionality analysis of the gastrointestinal tract microbiota. Gut 57: 1605–1615.
