Top

Published in:

Open Access 01-12-2017 | Research

Detection of enterotoxigenic Bacteroides fragilis in patients with ulcerative colitis

Authors: Samin Zamani, Sonia Hesam Shariati, Mohammad Reza Zali, Hamid Asadzadeh Aghdaei, Akram Sarabi Asiabar, Saied Bokaie, Bizhan Nomanpour, Leonardo Antonio Sechi, Mohammad Mehdi Feizabadi

Published in: Gut Pathogens | Issue 1/2017

Abstract

Purpose

Ulcerative colitis (UC) as a type of inflammatory bowel disease (IBD), presumed to occur as a consequence of increased immune responses to intestinal microbiota in genetically susceptible individuals. Enterotoxigenic Bacteroides fragilis (ETBF) strains are important intestinal bacteria that can be involved in IBD. The aim of this study was to design a quantitative assay for detection of B. fragilis and ETBF and also to find their association with UC.

Methods

Ninety-five biopsies were collected from patients with UC (n = 35) and with no IBD (nIBD, n = 60). All the specimens were cultured in Bacteroides bile esculin agar medium. Specific primers and probes were designed for quantitative real-time PCR (QRT-PCR) based on 16S rRNA and bft genes sequences of ETBF.

Results

The bft genes were detected in 51.4% of UC samples and 1.6% of nIBD samples, respectively. In UC patients, 37.1% of samples with diarrhea and 11.4% of samples without diarrhea, harbored the bft gene. Mean value of the number of ETBF with bft gene in UC and nIBD samples were 4.46 ×Ÿ 10² and 1.96, respectively. Likewise these result for 16S rRNA gene in UC and nIBD samples were 2.0 × 10³ and 8.4 × 10³, respectively.

Conclusions

There was no significant association between presence and numbers of 16S rRNA gene of B. fragilis and UC. ETBF was detected more in UC specimens and biopsies of UC patients with diarrhea than in the control group. These data demonstrated that ETBF is associated with development of UC and as a causative agent for the development of diarrhea in these patients.

Ley RE, Peterson DA, Gordon JI. Ecological and evolutionary forces shaping microbial diversity in the human intestine. Cell. 2006;124(4):837–48.CrossRefPubMed

Sartor RB. Genetics and environmental interactions shape the intestinal microbiome to promote inflammatory bowel disease versus mucosal homeostasis. Gastroenterology. 2010;139(6):1816–9.CrossRefPubMed

Lucke K, Miehlke S, Jacobs E, Schuppler M. Prevalence of Bacteroides and Prevotella spp. in ulcerative colitis. J Med Microbiol. 2006;55(5):617–24.CrossRefPubMed

Cummings JH, Macfarlane GT, Macfarlane S. Intestinal bacteria and ulcerative colitis. Curr Issues Intest Microbiol. 2003;4(1):9–20.PubMed

Sartor RB. Mechanisms of disease: pathogenesis of Crohn’s disease and ulcerative colitis. Nat Clin Pract Gastroenterol Hepatol. 2006;3(7):390–407.CrossRefPubMed

Khor B, Gardet A, Xavier RJ. Genetics and pathogenesis of inflammatory bowel disease. Nature. 2011;474(7351):307–17.CrossRefPubMedPubMedCentral

Campieri M, Gionchetti P. Bacteria as the cause of ulcerative colitis. Gut. 2001;48(1):132–5.CrossRefPubMedPubMedCentral

Zamani S, Zali MR, Aghdaei HA, Sechi LA, Niegowska M, Caggiu E, Keshavarz R, Mosavari N, Feizabadi MM. Mycobacterium avium subsp. paratuberculosis and associated risk factors for inflammatory bowel disease in Iranian patients. Gut Pathog. 2017;9(1):1.CrossRefPubMedPubMedCentral

van Kruiningen HJ. On the use of antibiotics in Crohn’s disease. J Clin Gastroenterol. 1995;20:310–6.CrossRefPubMed

10.

Shen BB, Qian JM. Intestinal flora and ulcerative colitis. Pract J Clin Med. 2008;5:002.

11.

Eckburg PBBE, Bernstein CN, Sargent M, Purdom EA, Relman DA. Molecular analysis of the colonic mucosal microbiota in patients with Crohn’s disease. Science. 2005;308:1635–8.CrossRefPubMedPubMedCentral

12.

Rehman A, Lepage P, Nolte A, Hellmig S, Schreiber S, et al. Transcriptional activity of the dominant gut mucosal microbiota in chronic inflammatory bowel disease patients. J Med Microbiol. 2010;59:1114–22.CrossRefPubMed

13.

Zoetendal EG, von Wright A, Vilpponen-Salmela T, Ben-Amor K, Akkermans AD, et al. Mucosa-associated bacteria in the human gastrointestinal tract are uniformly distributed along the colon and differ from the community recovered from feces. Appl Environ Microbiol. 2002;68:3401–7.CrossRefPubMedPubMedCentral

14.

Gophna U, Sommerfeld K, Gophna S, Doolittle WF, van Zanten SJV. Differences between tissue-associated intestinal microfloras of patients with Crohn’s disease and ulcerative colitis. J Clin Microbiol. 2006;44:4136–41.CrossRefPubMedPubMedCentral

15.

Wells CL, van de Westerlo EM, Jechorek RP, Feltis BA, Wilkins TD, Erlandsen SL. Bacteroides fragilis enterotoxin modulates epithelial permeability and bacterial internalization by HT-29 enterocytes. Gastroenterology. 1996;110(5):1429–37.CrossRefPubMed

16.

Prindiville TP, Sheikh RA, Cohen SH, Tang YJ, Cantrell MC, Silva J Jr. Bacteroides fragilis enterotoxin gene sequences in patients with inflammatory bowel disease. Emerg Infect Dis. 2000;6(2):171.CrossRefPubMedPubMedCentral

17.

Rabizadeh S, Rhee KJ, Wu S, Huso D, Gan CM, Golub JE, Sears CL. Enterotoxigenic Bacteroides fragilis: a potential instigator of colitis. Inflamm Bowel Dis. 2007;13(12):1475–83.CrossRefPubMedPubMedCentral

18.

Sack RB, Myers LL, Almeido-Hill J, Shoop DS, Bradbury WC, Reid R, Santosham M. Enterotoxigenic Bacteroides fragilis: epidemiologic studies of its role as a human diarrhoeal pathogen. J Diarrhoeal Dis Res. 1992;10:4–9.PubMed

19.

Pathela P, Hasan KZ, Roy E, Alam K, Huq F, Siddique AK, Sack RB. Enterotoxigenic Bacteroides fragilis-associated diarrhea in children 0–2 years of age in rural Bangladesh. J Infect Dis. 2005;191(8):1245–52.CrossRefPubMed

20.

Basset C, Holton J, Bazeos A, Vaira D, Bloom S. Are Helicobacter species and enterotoxigenic Bacteroides fragilis involved in inflammatory bowel disease? Dig Dis Sci. 2004;49(9):1425–32.CrossRefPubMed

21.

Dignass A, Eliakim R, Magro F, Maaser C, Chowers Y, Geboes K, et al. Second European evidence-based consensus on the diagnosis and management of ulcerative colitis part 1: definitions and diagnosis. J Crohns Colitis. 2012;6(10):965–90.CrossRefPubMed

22.

Adams PS. Data analysis and reporting. In: Dorak MT, editor. Real-time PCR. New York: Taylor and Francis; 2006. p. 39–61.

23.

Gismera CS, Aladren BS. Inflammatory bowel diseases: a disease(s) of modern times? Is incidence still increasing? World J Gastroenterol. 2008;14(36):5491–8.CrossRefPubMedPubMedCentral

24.

Daryani NE, Bashashati M, Aram S, Hashtroudi AA, Shakiba M, Sayyah A, et al. Pattern of relapses in Iranian patients with ulcerative colitis. A prospective study. J Gastrointest Liver Dis. 2006;15(4):355.

25.

Musso G, Gambino R, Cassander M. Interactions between gut microbiota and host metabolism predisposing to obesity and diabetes. Annu Rev Med. 2011;63:361–80.CrossRef

26.

Cáceres M, Zhang G, Weintraub A, Nord CE. Prevalence and antimicrobial susceptibility of enterotoxigenic Bacteroides fragilis in children with diarrhea in Nicaragua. Anaerobe. 2000;6:143–8.CrossRef

27.

Scotto d’Abusco AS, M Del Grosso and M Pantosti. Characterization of the enterotoxin gene of Bacteroides fragilis strains from different human sources, abstr. 3.106, p. 56. In: Abstracts of the 2nd world congress on anaerobic bacteria and infections. Boston: International Society of Anaerobic Bacteria; 1998.

28.

Sears CL, Islam S, Saha A, Arjumand M, Alam NH, Faruque AS, et al. Association of enterotoxigenic Bacteroides fragilis infection with inflammatory diarrhea. Clin Infect Dis. 2008;47:797–803.CrossRefPubMedPubMedCentral

29.

Boleij A, Hechenbleikner EM, Goodwin AC, Badani R, Stein EM, Lazarev MG, et al. The Bacteroides fragilis toxin gene is prevalent in the colon mucosa of colorectal cancer patients. Clin Infect Dis. 2015;60:208–15.CrossRefPubMed

30.

Rhee KJ, Wu S, Wu X, Huso DL, Karim B, Franco AA, et al. Induction of persistent colitis by a human commensal, enterotoxigenic Bacteroides fragilis, in wild-type C57BL/6 mice. Infect Immun. 2009;77:1708–18.CrossRefPubMedPubMedCentral

31.

Sears CL, Geis AL, Housseau F. Bacteroides fragilis subverts mucosal biology: from symbiont to colon carcinogenesis. J Clin Investig. 2014;124:4166–72.CrossRefPubMedPubMedCentral

32.

Herrinton LJ, Liu L, Levin TR, Allison JE, Lewis JD, Velayos F. Incidence and mortality of colorectal adenocarcinoma in persons with inflammatory bowel disease from 1998 to 2010. Gastroenterology. 2012;143:382–9.CrossRefPubMed

33.

Gillen CD, Walmsley RS, Prior P, Andrews HA, Allan RN. Ulcerative colitis and Crohn’s disease: a comparison of the colorectal cancer risk in extensive colitis. Gut. 1994;35:1590–2.CrossRefPubMedPubMedCentral

34.

Hartley MG, Hudson MJ, Swarbrick ET, Hill MJ, Gent AE, Hellier MD, Grace RH. The rectal mucosa-associated microflora in patients with ulcerative colitis. J Med Microbiol. 1992;36:96–103.CrossRefPubMed

35.

Kato N, Liu CX, Kato H, Watanabe K, Tanaka Y, Yamamoto T, Suzuki K, Ueno K. A new subtype of the metalloprotease toxin gene and the incidence of the three bft subtypes among Bacteroides fragilis isolates in Japan. FEMS Microbiol Lett. 2000;182(1):171–6.CrossRefPubMed

36.

Avila-Campos MJ, Liu C, Song Y, Rowlinson MC, Finegold SM. Determination of bft gene subtypes in Bacteroides fragilis clinical isolates. J Clin Microbiol. 2007;45(4):1336–8.CrossRefPubMedPubMedCentral

37.

Akhi MT, Seifi SJ, Asgharzadeh M, Rezaee MA, Oskuei SA, Pirzadeh T, Memar MY, Alizadeh N, Sofla HS. Role of Enterotoxigenic Bacteroides fragilis in Children Less Than 5 Years of Age With Diarrhea in Tabriz, Iran. Jundishapur journal of microbiology. 2016;9(6):e32163.

38.

Zhou Y, Zhi F. Lower level of bacteroides in the gut microbiota is associated with inflammatory bowel disease: a meta-analysis. Biomed Res Int. 2016;24:2016.

39.

Takaishi H, Matsuki T, Nakazawa A, Takada T, Kado S, Asahara T, Kamada N, Sakuraba A, Yajima T, Higuchi H, Inoue N. Imbalance in intestinal microflora constitution could be involved in the pathogenesis of inflammatory bowel disease. Int J Med Microbiol. 2008;298(5):463–72.CrossRefPubMed

Title: Detection of enterotoxigenic Bacteroides fragilis in patients with ulcerative colitis
Authors: Samin Zamani
Sonia Hesam Shariati
Mohammad Reza Zali
Hamid Asadzadeh Aghdaei
Akram Sarabi Asiabar
Saied Bokaie
Bizhan Nomanpour
Leonardo Antonio Sechi
Mohammad Mehdi Feizabadi
Publication date: 01-12-2017
Publisher: BioMed Central
Published in: Gut Pathogens / Issue 1/2017
Electronic ISSN: 1757-4749
DOI: https://doi.org/10.1186/s13099-017-0202-0

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Detection of enterotoxigenic Bacteroides fragilis in patients with ulcerative colitis

Abstract

Purpose

Methods

Results

Conclusions

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2017

Existence of a novel qepA variant in quinolone resistant Escherichia coli from aquatic habitats of Bangladesh

Comparison of clinical and immunological findings in gnotobiotic piglets infected with Escherichia coli O104:H4 outbreak strain and EHEC O157:H7

Prevalence of pks-positive Escherichia coli in Japanese patients with or without colorectal cancer

Oral administration of Clostridium butyricum CGMCC0313-1 inhibits β-lactoglobulin-induced intestinal anaphylaxis in a mouse model of food allergy

Attenuation of Helicobacter pylori-induced gastric inflammation by prior cag − strain (AM1) infection in C57BL/6 mice

Outer membrane phospholipase A’s roles in Helicobacter pylori acid adaptation

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page