Microbial Ecology

, Volume 61, Issue 3, pp 693–703 | Cite as

Intestinal Microbiota Was Assessed in Cirrhotic Patients with Hepatitis B Virus Infection

Intestinal Microbiota of HBV Cirrhotic Patients
  • Haifeng Lu
  • Zhongwen Wu
  • Wei Xu
  • Jiezuan Yang
  • Yunbo Chen
  • Lanjuan Li
Host Microbe Interaction

Abstract

To unravel the profile of intestinal microecological parameters in Chinese patients with asymptomatic carriage of hepatitis B virus (HBV), chronic hepatitis B, decompensated HBV cirrhosis, and health controls and to establish their correlation with liver disease progression, we performed quantitative PCR and immunological techniques to investigate fecal parameters, including population of fecal predominant bacteria and the abundance of some virulence genes derived from Escherichia coli, Bacteroides fragilis, Clostridium difficile, and Clostridium perfringens in fecal crude DNA and some immunological parameters in extracts of all fecal samples. Data analysis indicated that 16S rRNA gene copy numbers for Faecalibacterium prausnitzii, Enterococcus faecalis, Enterobacteriaceae, bifidobacteria, and lactic acid bacteria (Lactobacillus, Pediococcus, Leuconostoc, and Weissella) showed marked variation in the intestine of HBV cirrhotic patients. The Bifidobacteria/Enterobacteriaceae (B/E) ratio, which may indicate microbial colonization resistance of the bowel, was decreased significantly in turn from 1.15 ± 0.11 in healthy controls, 0.99 ± 0.09 in asymptomatic carriers, and 0.76 ± 0.08 in patients with chronic hepatitis B to 0.64 ± 0.09 in patients with decompensated HBV cirrhosis (for all, P < 0.01). This suggests that B/E ratio is useful for following the level of intestinal microecological disorder in the course of liver disease progression. The data for virulence gene abundance suggested increased diversity of virulence factors during liver disease progression. Fecal secretory IgA and tumor necrosis factor-α in decompensated HBV cirrhotic patients were present at higher levels than in other groups, which indicates that a complicated autoregulatory system tries to achieve a new intestinal microecological balance.

References

  1. 1.
    Barbut F, Gariazzo B, Bonne L, Lalande V, Burghoffer B, Luiuz R, Petit JC (2007) Clinical features of Clostridium difficile-associated infections and molecular characterization of strains: results of a retrospective study, 2000–2004. Infect Control Hosp Epidemiol 28:131–139PubMedCrossRefGoogle Scholar
  2. 2.
    Bartosch S, Fite A, Macfarlane GT, McMurdo ME (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 70:3575–3581PubMedCrossRefGoogle Scholar
  3. 3.
    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 40:28–37PubMedCrossRefGoogle Scholar
  4. 4.
    Beckebaum S, Malago M, Dirsch O, Cicinnati VR, Trippler M, Lampertico P, Lama N, Treichel U, Gerken G, Broelsch CE (2003) Efficacy of combined lamivudine and adefovir dipivoxil treatment for severe HBV graft reinfection after living donor liver transplantation. Clin Transplant 17:554–559PubMedCrossRefGoogle Scholar
  5. 5.
    Begley M, Gahana CGM, Hill C (2005) The interaction between bacteria and bile. FEMS Microbiol Rev 29:625–651PubMedCrossRefGoogle Scholar
  6. 6.
    Blum S, Schiffrin EJ (2003) Intestinal microflora and homeostasis of the mucosal immune response: implications for probiotic bacteria? Curr Issues Intest Microbiol 4:53–60PubMedGoogle Scholar
  7. 7.
    Boullier S, Tanguy M, Kadaoui KA, Caubet C, Sansonetti P, Corthesy B, Phalipon A (2009) Secretory IgA-mediated neutralization of Shigella flexneri prevents intestinal tissue destruction by down-regulating inflammatory circuits. J Immunol 183:5879–5885PubMedCrossRefGoogle Scholar
  8. 8.
    Brenchley JM, Price DA, Schacker TW, Asher TE, Silvestri G, Rao S, Kazzaz Z, Bornstein E, Lambotte O, Altmann D, Blazar BR, Rodriguez B, Teixeira-Johnson L, Landay A, Martin JN, Hecht FM, Picker LJ, Lederman MM, Deeks SG, Douek DC (2006) Microbial translocation is a cause of systemic immune activation in chronic HIV infection. Nat Med 12:1365–1371PubMedCrossRefGoogle Scholar
  9. 9.
    Castelnau C, Le Gal F, Ripault MP, Gordien E, Martinot-Peignoux M, Boyer N, Pham BN, Maylin S, Bedossa P, Deny P, Marcellin P, Gault E (2006) Efficacy of peginterferon alpha-2b in chronic hepatitis delta: relevance of quantitative RT-PCR for follow-up. Hepatology (Baltimore, Md) 44:728–735Google Scholar
  10. 10.
    Chesta J, Silva M, Thompson L, del Canto E, Defilippi C (1991) Bacterial overgrowth in small intestine in patients with liver cirrhosis. Rev Méd Chile 119:626–632PubMedGoogle Scholar
  11. 11.
    Collins MD, Lawson PA, Willems A, Cordoba JJ, Fernandez-Garayzabal J, Garcia P, Cai J, Hippe H, Farrow JA (1994) The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. Int J Syst Bacteriol 44:812–826PubMedCrossRefGoogle Scholar
  12. 12.
    Cummings JH, Antoine JM, Azpiroz F, Bourdet-Sicard R, Brandtzaeg P, Calder PC, Gibson GR, Guarner F, Isolauri E, Pannemans D, Shortt C, Tuijtelaars S, Watzl B (2004) PASSCLAIM—gut health and immunity. Eur J Nutr 43(Suppl 2):II118–II173PubMedCrossRefGoogle Scholar
  13. 13.
    Delacroix DL, Reynaert M, Pauwels S, Geubel AP, Vaerman JP (1982) High serum levels of secretory IgA in liver disease: possible liver origin of the circulating secretory component. Dig Dis Sci 27:333–340PubMedCrossRefGoogle Scholar
  14. 14.
    Floch MH, Katz J, Conn HO (1970) Qualitative and quantitative relationships of the fecal flora in cirrhotic patients with portal systemic encephalopathy and following portacaval anastomosis. Gastroenterology 59:70–75PubMedGoogle Scholar
  15. 15.
    Fukushima H, Tsunomori Y, Seki R (2003) Duplex real-time SYBR green PCR assays for detection of 17 species of food- or waterborne pathogens in stools. J Clin Microbiol 41:5134–5146PubMedCrossRefGoogle Scholar
  16. 16.
    Guarner F, Malagelada JR (2003) Gut flora in health and disease. Lancet 361:512–519PubMedCrossRefGoogle Scholar
  17. 17.
    Haarman M, Knol J (2006) Quantitative real-time PCR analysis of fecal Lactobacillus species in infants receiving a prebiotic infant formula. Appl Environ Microbiol 72:2359–2365PubMedCrossRefGoogle Scholar
  18. 18.
    Han DW (2002) Intestinal endotoxemia as a pathogenetic mechanism in liver failure. World J Gastroenterol 8:961–965PubMedGoogle Scholar
  19. 19.
    Hilali F, Ruimy R, Saulnier P, Barnabe C, Lebouguenec C, Tibayrenc M, Andremont A (2000) Prevalence of virulence genes and clonality in Escherichia coli strains that cause bacteremia in cancer patients. Infect Immun 68:3983–3989PubMedCrossRefGoogle Scholar
  20. 20.
    Hodgson HJ (1985) Gut–liver interactions in the IgA system. Scand J Gastroenterol Suppl 114:39–44PubMedCrossRefGoogle Scholar
  21. 21.
    Joishy M, Davies I, Ahmed M, Wassel J, Davies K, Sayers A, Jenkins H (2009) Fecal calprotectin and lactoferrin as noninvasive markers of pediatric inflammatory bowel disease. J Pediatr Gastroenterol Nutr 48:48–54PubMedCrossRefGoogle Scholar
  22. 22.
    Jones J, Loftus EV Jr, Panaccione R, Chen LS, Peterson S, McConnell J, Baudhuin L, Hanson K, Feagan BG, Harmsen SW, Zinsmeister AR, Helou E, Sandborn WJ (2008) Relationships between disease activity and serum and fecal biomarkers in patients with Crohn’s disease. Clin Gastroenterol Hepatol 6:1218–1224PubMedCrossRefGoogle Scholar
  23. 23.
    Kapel N, Roman C, Caldari D, Sieprath F, Canioni D, Khalfoun Y, Goulet O, Ruemmele FM (2005) Fecal tumor necrosis factor-alpha and calprotectin as differential diagnostic markers for severe diarrhea of small infants. J Pediatr Gastroenterol Nutr 41:396–400PubMedCrossRefGoogle Scholar
  24. 24.
    Lamb CA, Mohiuddin MK, Gicquel J, Neely D, Bergin FG, Hanson JM, Mansfield JC (2009) Faecal calprotectin or lactoferrin can identify postoperative recurrence in Crohn’s disease. Br J Surg 96:663–674PubMedCrossRefGoogle Scholar
  25. 25.
    Ma TY, Iwamoto GK, Hoa NT, Akotia V, Pedram A, Boivin MA, Said HM (2004) TNF-alpha-induced increase in intestinal epithelial tight junction permeability requires NF-kappa B activation. Am J Physiol 286:G367–G376CrossRefGoogle Scholar
  26. 26.
    Macpherson AJ, Uhr T (2004) Induction of protective IgA by intestinal dendritic cells carrying commensal bacteria. Science (New York, NY) 303:1662–1665CrossRefGoogle Scholar
  27. 27.
    Mariat D, Firmesse O, Levenez F, Guimaraes V, Sokol H, Dore J, Corthier G, Furet JP (2009) The Firmicutes/Bacteroidetes ratio of the human microbiota changes with age. BMC Microbiol 9:123PubMedCrossRefGoogle Scholar
  28. 28.
    Martini GA, Phear EA, Ruebner B, Sherlock S (1957) The bacterial content of the small intestine in normal and cirrhotic subjects: relation to methionine toxicity. Clin Sci (Lond) 16:35–51Google Scholar
  29. 29.
    Miettinen TA (1972) Lipid absorption, bile acids, and cholesterol metabolism in patients with chronic liver disease. Gut 13:682–689PubMedCrossRefGoogle Scholar
  30. 30.
    Moriya K, Sumi N, Nakao M, Yamazaki M, Saito M, Ito K (2000) The prevalence of virulence-related genes, eaeA, aggR and astA, of localized and aggregative-adherent Escherichia coli (EPEC and EAggEC) in healthy children and age-matched patients with diarrhea. Kansenshogaku Zasshi 74:134–142PubMedGoogle Scholar
  31. 31.
    Murray WR, Blackwood A, Calman KC, MacKay C (1980) Faecal bile acids and clostridia in patients with breast cancer. Br J Cancer 42:856–860PubMedCrossRefGoogle Scholar
  32. 32.
    O’Sullivan DJ (2008) Genomics can advance the potential for probiotic cultures to improve liver and overall health. Curr Pharm Des 14:1376–1381PubMedCrossRefGoogle Scholar
  33. 33.
    Palma P, Mihaljevic N, Hasenberg T, Keese M, Koeppel TA (2007) Intestinal barrier dysfunction in developing liver cirrhosis: an in vivo analysis of bacterial translocation. Hepatol Res 37:6–12PubMedCrossRefGoogle Scholar
  34. 34.
    Pelletier G, Briantais MJ, Buffet C, Pillot J, Etienne JP (1982) Serum and intestinal secretory IgA in alcoholic cirrhosis of the liver. Gut 23:475–480PubMedCrossRefGoogle Scholar
  35. 35.
    Ramachandran A, Balasubramanian KA (2001) Intestinal dysfunction in liver cirrhosis: its role in spontaneous bacterial peritonitis. J Gastroenterol Hepatol 16:607–612PubMedCrossRefGoogle Scholar
  36. 36.
    Scholtens PA, Alliet P, Raes M, Alles MS, Kroes H, Boehm G, Knippels LM, Knol J, Vandenplas Y (2008) Fecal secretory immunoglobulin A is increased in healthy infants who receive a formula with short-chain galacto-oligosaccharides and long-chain fructo-oligosaccharides. J Nutr 138:1141–1147PubMedGoogle Scholar
  37. 37.
    Sharma N, Chaudhry R (2006) Rapid detection of enterotoxigenic Bacteroides fragilis in diarrhoeal faecal samples. Indian J Med Res 124:575–582PubMedGoogle Scholar
  38. 38.
    Si JM, Yu YC, Fan YJ, Chen SJ (2004) Intestinal microecology and quality of life in irritable bowel syndrome patients. World J Gastroenterol 10:1802–1805PubMedGoogle Scholar
  39. 39.
    Sokol H, Pigneur B, Watterlot L, Lakhdari O, Bermudez-Humaran LG, Gratadoux JJ, Blugeon S, Bridonneau C, Furet JP, Corthier G, Grangette C, Vasquez N, Pochart P, Trugnan G, Thomas G, Blottiere HM, Dore J, Marteau P, Seksik P, Langella P (2008) Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patients. Proc Natl Acad Sci USA 105:16731–16736PubMedCrossRefGoogle Scholar
  40. 40.
    Song Y, Liu C, Finegold SM (2004) Real-time PCR quantitation of clostridia in feces of autistic children. Appl Environ Microbiol 70:6459–6465PubMedCrossRefGoogle Scholar
  41. 41.
    Suzuki K, Meek B, Doi Y, Muramatsu M, Chiba T, Honjo T, Fagarasan S (2004) Aberrant expansion of segmented filamentous bacteria in IgA-deficient gut. Proc Natl Acad Sci U S A 101(7):1981–1986PubMedCrossRefGoogle Scholar
  42. 42.
    Suzuki MT, Taylor LT, DeLong EF (2000) Quantitative analysis of small-subunit rRNA genes in mixed microbial populations via 5′-nuclease assays. Appl Environ Microbiol 66:4605–4614PubMedCrossRefGoogle Scholar
  43. 43.
    Toprak NU, Yagci A, Gulluoglu BM, Akin ML, Demirkalem P, Celenk T, Soyletir G (2006) A possible role of Bacteroides fragilis enterotoxin in the aetiology of colorectal cancer. Clin Microbiol Infect 12:782–786PubMedGoogle Scholar
  44. 44.
    Usein CR, Damian M, Tatu-Chitoiu D, Capusa C, Fagaras R, Tudorache D, Nica M, Le Bouguenec C (2001) Prevalence of virulence genes in Escherichia coli strains isolated from Romanian adult urinary tract infection cases. J Cell Mol Med 5:303–310PubMedCrossRefGoogle Scholar
  45. 45.
    van der Waaij D (1992) History of recognition and measurement of colonization resistance of the digestive tract as an introduction to selective gastrointestinal decontamination. Epidemiol Infect 109:315–326PubMedCrossRefGoogle Scholar
  46. 46.
    Williams R (2007) Review article: bacterial flora and pathogenesis in hepatic encephalopathy. Aliment Pharmacol Ther 25(Suppl 1):17–22PubMedCrossRefGoogle Scholar
  47. 47.
    Yan F, Polk DB (2004) Commensal bacteria in the gut: learning who our friends are. Curr Opin Gastroenterol 20:565–571PubMedCrossRefGoogle Scholar
  48. 48.
    Yu H, Cao ZC, Geng JY, He ZG, Wang ZM, Sun XY, Wang ZH (2005) Effect of interferon alpha on liver fibrosis in patients with chronic hepatitis B. Di 1 Jun Yi Da Xue Xue Bao (Acad J First Med College PLA) 25:1409–1412Google Scholar
  49. 49.
    Zeuzem S (2000) Gut-liver axis. Int J Colorectal Dis 15:59–82PubMedCrossRefGoogle Scholar
  50. 50.
    Zhang W, Zhao M, Ruesch L, Omot A, Francis D (2007) Prevalence of virulence genes in Escherichia coli strains recently isolated from young pigs with diarrhea in the US. Vet Microbiol 123:145–152PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Haifeng Lu
    • 1
  • Zhongwen Wu
    • 1
  • Wei Xu
    • 1
  • Jiezuan Yang
    • 1
  • Yunbo Chen
    • 1
  • Lanjuan Li
    • 1
  1. 1.State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, College of MedicineZhejiang UniversityHangzhouPeople’s Republic of China

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