Changes of Fecal Bifidobacterium Species in Adult Patients with Hepatitis B Virus-Induced Chronic Liver Disease

The beneficial effects of Bifidobacteria on health have been widely accepted. Patients with chronic liver disease have varying degrees of intestinal microflora imbalance with a decrease of total Bifidobacterial counts. Since different properties have been attributed to different Bifidobacterium species and there is no information available for the detailed changes in the genus Bifidobacterium in patients with chronic liver disease heretofore, it is meaningful to investigate the structure of this bacterium at the species level in these patients. The aim of this study was to characterize the composition of intestinal Bifidobacterium in patients with hepatitis B virus-induced chronic liver disease. Nested-PCR-based denaturing gradient gel electrophoresis (PCR-DGGE), clone library, and real-time quantitative PCR were performed on the fecal samples of 16 patients with chronic hepatitis B (CHB patients), 16 patients with hepatitis B virus-related cirrhosis (HBV cirrhotics), and 15 healthy subjects (Controls). Though there was no significant difference in the diversity among the three groups (P = 0.196), Bifidobacterium dentium seems to be specifically enhanced in patients as the PCR-DGGE profiles showed, which was further validated by clone library and real-time quantitative PCR. In contrast to the B. dentium, Bifidobacteriumcatenulatum/Bifidobacterium pseudocatenulatum were detected less frequently in the predominant profile and by quantitative PCR in HBV cirrhotics than in the controls, and the level of this species was also significantly different between these two groups (P = 0.023). Although having no quantitative difference among the three groups, Bifidobacterium longum was less commonly detected in HBV cirrhotics than in CHB patients and Controls by quantitative PCR (P = 0.011). Thus, the composition of intestinal Bifidobacterium was deeply altered in CHB and HBV cirrhotic patients with a shift from beneficial species to opportunistic pathogens. The results provide further insights into the dysbiosis of the intestinal microbiota in patients with hepatitis B virus-induced chronic liver disease and might potentially serve as guidance for the probiotics interventions of these diseases.

[1]  K. Tateda,et al.  Oral administration of Bifidobacterium longum prevents gut‐derived Pseudomonas aeruginosa sepsis in mice , 2008, Journal of applied microbiology.

[2]  T. Mitsuoka.  Bifidobacteria and their role in human health , 1990, Journal of Industrial Microbiology.

[3]  Y. Sanz,et al.  Imbalances in faecal and duodenal Bifidobacterium species composition in active and non-active coeliac disease , 2008, BMC Microbiology.

[4]  O. Korshunova,et al.  Molecular Genetic Study of Species and Strain Variability in Bifidobacteria Population in Intestinal Microflora of Breast-Fed Infants and Their Mothers , 2010, Bulletin of Experimental Biology and Medicine.

[5]  Shu-sen Zheng,et al.  Protective role of supplement with foreign Bifidobacterium and Lactobacillus in experimental hepatic ischemia‐reperfusion injury , 2006, Journal of gastroenterology and hepatology.

[6]  K. Michaelsen,et al.  Case Study of the Distribution of Mucosa-Associated Bifidobacterium Species, Lactobacillus Species, and Other Lactic Acid Bacteria in the Human Colon , 2003, Applied and Environmental Microbiology.

[7]  Roger Williams,et al.  The intestinal flora and bacterial infection in cirrhosis. , 2006, Journal of hepatology.

[8]  V. Rada,et al.  Cross-talk of human gut with bifidobacteria. , 2009, Nutrition reviews.

[9]  J. Handelsman,et al.  Introducing DOTUR, a Computer Program for Defining Operational Taxonomic Units and Estimating Species Richness , 2005, Applied and Environmental Microbiology.

[10]  S. Salminen,et al.  Quantitative assessment of faecal bifidobacterial populations by real‐time PCR using lanthanide probes , 2006, Journal of applied microbiology.

[11]  D. Sinderen,et al.  Microbiomic analysis of the bifidobacterial population in the human distal gut , 2009, The ISME Journal.

[12]  R. Wiest,et al.  Gut microflora in the pathogenesis of the complications of cirrhosis. , 2004, Best practice & research. Clinical gastroenterology.

[13]  L. Hoyles,et al.  Examination of faecal Bifidobacterium populations in breast- and formula-fed infants during the first 18 months of life. , 2010, Microbiology.

[14]  N. Maeda,et al.  Reduction of vitamin K concentration by salivary Bifidobacterium strains and their possible nutritional competition with Porphyromonas gingivalis , 2007, Journal of applied microbiology.

[15]  C. Huh,et al.  Hepatoprotective effect of lactic acid bacteria, inhibitors of β-glucuronidase production against intestinal microflora , 2005, Archives of pharmacal research.

[16]  P. Adamski,et al.  Mucosal bacterial microflora and mucus layer thickness in adolescents with inflammatory bowel disease. , 2009, World journal of gastroenterology.

[17]  M. Romond,et al.  Bifidobacteria and human health: regulatory effect of indigenous bifidobacteria on Escherichia coli intestinal colonization. , 1997, Anaerobe.

[18]  Peter Chung,et al.  Transmission of Diverse Oral Bacteria to Murine Placenta: Evidence for the Oral Microbiome as a Potential Source of Intrauterine Infection , 2010, Infection and Immunity.

[19]  A. Margolles,et al.  Distinct Bifidobacterium strains drive different immune responses in vitro. , 2010, International journal of food microbiology.

[20]  Wei Xu,et al.  Intestinal Microbiota Was Assessed in Cirrhotic Patients with Hepatitis B Virus Infection , 2011, Microbial Ecology.

[21]  Qiang Liu,et al.  Diversity and abundance of the bacterial 16S rRNA gene sequences in forestomach of alpacas (Lama pacos) and sheep (Ovis aries). , 2010, Anaerobe.

[22]  Maria Saarela,et al.  Bifidobacterial Diversity in Human Feces Detected by Genus-Specific PCR and Denaturing Gradient Gel Electrophoresis , 2001, Applied and Environmental Microbiology.

[23]  M. Fenlon,et al.  The Isolation of Bifidobacteria from Occlusal Carious Lesions in Children and Adults , 2009, Caries Research.

[24]  R. Tanaka,et al.  Distribution of Bifidobacterial Species in Human Intestinal Microflora Examined with 16S rRNA-Gene-Targeted Species-Specific Primers , 1999, Applied and Environmental Microbiology.

[25]  M. Samsom,et al.  Lower Bifidobacteria counts in both duodenal mucosa-associated and fecal microbiota in irritable bowel syndrome patients. , 2009, World journal of gastroenterology.

[26]  J. Fujimoto,et al.  Quantitative PCR with 16S rRNA-Gene-Targeted Species-Specific Primers for Analysis of Human Intestinal Bifidobacteria , 2004, Applied and Environmental Microbiology.

[27]  G. Holtrop,et al.  Effect of inulin on the human gut microbiota: stimulation of Bifidobacterium adolescentis and Faecalibacterium prausnitzii , 2008, British Journal of Nutrition.

[28]  Zhuang,et al.  Guideline on prevention and treatment of chronic hepatitis B in China (2005). , 2007, Chinese medical journal.

[29]  M. Kamm,et al.  Bacterial and fungal microbiota in relation to probiotic therapy (VSL#3) in pouchitis , 2006, Gut.

[30]  D. Lavanchy,et al.  Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures , 2004, Journal of viral hepatitis.

[31]  Y. Chen,et al.  Changes in intestinal microflora in patients with chronic severe hepatitis. , 2001, Chinese medical journal.

[32]  L. Zoitopoulos,et al.  Isolation and Identification of Bifidobacteriaceae from Human Saliva , 2008, Applied and Environmental Microbiology.

[33]  D. Beighton,et al.  Resistance to Acidic Environments of Caries-Associated Bacteria: Bifidobacterium dentium and Bifidobacterium longum , 2010, Caries Research.

[34]  R. Knight,et al.  Evolution of Mammals and Their Gut Microbes , 2008, Science.

[35]  Marcus J. Claesson,et al.  The Bifidobacterium dentium Bd1 Genome Sequence Reflects Its Genetic Adaptation to the Human Oral Cavity , 2009, PLoS genetics.

[36]  R. Pugh,et al.  Transection of the oesophagus for bleeding oesophageal varices , 1973, The British journal of surgery.

[37]  G. Garcia‐Tsao,et al.  Probiotics and Liver Disease , 2008, Journal of clinical gastroenterology.

[38]  J. Steiner,et al.  Assessment of microbial diversity along the feline intestinal tract using 16S rRNA gene analysis. , 2008, FEMS microbiology ecology.

[39]  R. Mackie,et al.  Denaturing Gradient Gel Electrophoresis Analysis of 16S Ribosomal DNA Amplicons To Monitor Changes in Fecal Bacterial Populations of Weaning Pigs after Introduction of Lactobacillus reuteri Strain MM53 , 2000, Applied and Environmental Microbiology.

[40]  Lanjuan Li,et al.  Effects of gut microflora on hepatic damage after acute liver injury in rats. , 2010, The Journal of trauma.

[41]  H. Zhao,et al.  Intestinal microflora in patients with liver cirrhosis. , 2004, Chinese journal of digestive diseases.

[42]  P. Bork,et al.  A human gut microbial gene catalogue established by metagenomic sequencing , 2010, Nature.

[43]  B. Roe,et al.  A core gut microbiome in obese and lean twins , 2008, Nature.

[44]  M. Pirlich,et al.  Gastrointestinal tract in liver disease: which organ is sick? , 2008, Current opinion in clinical nutrition and metabolic care.

[45]  Y. Sanz,et al.  Differences in faecal bacterial communities in coeliac and healthy children as detected by PCR and denaturing gradient gel electrophoresis. , 2007, FEMS immunology and medical microbiology.

[46]  M. Malaguarnera,et al.  Bifidobacterium longum with Fructo-Oligosaccharide (FOS) Treatment in Minimal Hepatic Encephalopathy: A Randomized, Double-Blind, Placebo-Controlled Study , 2007, Digestive Diseases and Sciences.

[47]  E. Zoetendal,et al.  Isolation of Bifidobacteria from Breast Milk and Assessment of the Bifidobacterial Population by PCR-Denaturing Gradient Gel Electrophoresis and Quantitative Real-Time PCR , 2008, Applied and Environmental Microbiology.

[48]  S. Delgado,et al.  Bifidobacterial Diversity Determined by Culturing and by 16S rDNA Sequence Analysis in Feces and Mucosa from Ten Healthy Spanish Adults , 2006, Digestive Diseases and Sciences.