Mycobacterium avium subsp. paratuberculosis, etiological factor in Crohn’s disease?

Authors: B. Fixa 1,5;  I. Pavlík 4;  J. Bedrna 2;  O. Komárková 1;  Z. Nožička 3;  I. Slaná 4;  P. Králík 4;  T. Vaňásek 5;  M. Volfová 5
Authors‘ workplace: II. interní klinika, LF UK a FN Hradec Králové 1;  Chirurgická klinika, LF UK a FN Hradec Králové 2;  Fingerlandův ústav patologické anatomie, LF UK a FN Hradec Králové 3;  Výzkumný ústav veterinárního lékařství, v. v. i., Brno 4;  Hepato-gastroenterologie s. r. o., Hradec Králové 5
Published in: Gastroent Hepatol 2010; 64(2): 7-13
Category: Original Article


The etiology of Crohn’s disease remains open. The significance of Mycobacterium avium subsp. paratuberculosis (MAP) in the development of Crohn’s Disease has for a long time been influenced by the great difficulties associated with the cultivation procedure. The goal of our study was to determine the incidence of MAP by the new highly sensitive PCR method (hsPCR) for the detection of specific DNA fragment IS900 and together with reports in the literature, to assess the position of MAP in the etiology of Crohn’s disease. Material and methods: Small specimens taken from the operated gut of 106 patients with Crohn’s disease were examined for the presence of MAP by a highly sensitive PCR method. Resection of terminal ileum and cecum – primary operations were performed in 61 patients, reoperation in 32 patients, dominant operation of the colon in 11 patients, resection of the jejunum in 2 patients. The results from the involved pathological tissue were compared with the results from macroscopically unaffected tissue (controls). Results: In affected tissue, MAP was present in 58 patients (54.7 %). In 14 of these patients (13.2 %), MAP was also found in control specimens. In 4 patients (3.8 %) MAP was found only in unaffected tissue. In total, MAP was found in 62 patients (58.5 %). Conclusion: A significantly higher incidence of MAP found in the affected tissue of patients with Crohn’s disease in comparison with unaffected tissue supports the view of the possible role of MAP in the etiology of Crohn’s disease.

Key words:
Crohn’s disease – Mycobacterium avium subsp. paratuberculosis – paratuberculosis – Johne‘s disease


1. Crohn BB, Ginzburg L, Oppenheimer GD. Regional ileitis: a pathologic and clinical entity. JAMA 1932; 99: 1323–1329.

2. Dalziel TK. Chronic intestinal enteritis. BMJ 1913; 2: 1068–1069.

3. Thayer WR, Coutu JA, Chiodini RJ et al. Possible role of Mycobacteria to inflammatory bowel disease. Dig Dis Sci 1984; 29(12): 1080–1085.

4. Chiodini RJ, Van Kruiningen HJ, Thayer WR et al. Possible role of mycobacteria in inflammatory bowel disease. I. An unclassified Mycobacterium species isolated from patients with Crohn’s disease. Dig Dis Sci 1984; 29(12): 1073–1079.

5. Fixa B, Komárková O, Bedrna J et al. Crohnova choroba a Mycobacterium avium subspecies paratuberculosis. Čes a Slov Gastroent 2000; 54(5): 56–61.

6. Ayele WY, Bartos M, Svastova P et al. Distribution of Mycobacterium avium subsp. paratuberculosis in organs of naturally infected bull-calves and breeding bulls. Vet Microbiol 2004; 103(3–4): 209–217.

7. Pavlík I, Bejčková L, Pavlas M et al. Characterization by restriction endonuclease analysis and DNA hybridization using IS900 of bovine, ovine, caprine and human dependent strains, of Mycobacterium paratuberculosis isolated in various localities. Vet Microbiol 1995; 45(4): 311–318.

8. Pavlik I, Horvathova A, Dvorska L et al. Standardisation of restriction fragment length polymorphism for Mycobacterium avium subspecies paratuberculosis. J Microbiol Meth 1999; 38(1–2): 155–167.

9. Sanderson JD, Moss MT, Tizard ML et al. Mycobacterium paratuberculosis DNA in Crohn’s disease tissue. Gut 1992; 33(7): 890–896.

10. Dell’Isola B, Poyart C, Goulet O et al. Detection of Mycobacterium paratuberculosis by polymerase chain reaction in children with Crohn’s disease. J Infect Dis 1994; 169(2): 449–451.

11. Lisby G, Andersen J, Engbaek K et al. Mycobacterium paratuberculosis in intestinal tissue from patients with Crohn’s disease demonstrated by a nested primer polymerase chain reaction. Scand J Gastroenterol 1994; 29(10): 923–929.

12. Fidler HM, Thurrell W, Johnson NM et al. Specific detection of Mycobacterium paratuberculosis DNA associated with granulomatous tissue in Crohn’s disease. Gut 1994; 35(4): 506–510.

13. Suenaga K, Yokoyama Y, Nishimori I et al. Serum antibodies to Mycobacterium paratuberculosis in patients with Crohn’s disease. Dig Dis Sci 1999; 44(6): 1202–1207.

14. Rowbotham DS, Mapstone NP, Trejdosiewicz LK et al. Mycobacterium paratuberculosis DNA not detected in Crohn’s disease tissue by fluorescent polymerase chain reaction. Gut 1995; 37: 660–667.

15. Dumonceau JM, Van Bossum A, Adler M et al. No Mycobacterium paratuberculosis found in Crohn’s disease using the polymerase chain reaction. Dige Dis Sci 1996; 41(2): 421–426.

16. Mishina D, Kastel P, Brown ST et al. On the etiology of Crohn disease. Proc Nat Acad Sci - US 1996; 93(18): 9816–9820.

17. Del Prete R, Quaranta M, Lippolis A et al. Detection of Mycobacterium paratuberculosis in stool samples of patients with inflammatory bowel disease by IS900-based PCR and colorimetric detection of amplified DNA. J Microbiol Methods 1998; 33: 105–114.

18. Collins MT, Lisby G, Moser C et al. Results of multiple diagnostic tests for Mycobacterium avium subsp. paratuberculosis in patients with inflammatory bowel disease and in controls. J Clin Microbiol 2000; 38(12): 4373–4381.

19. Autschbach F, Eisold S, Hinz U et al. High prevalence of Mycobacterium avium subspecies paratuberculosis IS900 DNA in gut tissues from individuals with Crohn’s disease. Gut 2005; 54(7): 944–949.

20. Bull TJE, McMinn EJ, Sidi-Boumedine K et al. Detection and verification of Mycobacterium avium subsp. paratuberculosis in fresh ileocolonic mucosal biopsy specimens from individuals with and without Crohn’s disease. J Clin Microbiol 2003; 41(7): 2915–2923.

21. Sechi LA, Scanu AM, Molicotti P et al. Detection and isolation of Mycobacterium avium subspecies paratuberculosis from intestinal mucosal biopsies of patients with and without Crohn’s disease in Sardinia. Am J Gastroenterol 2005; 100(7): 1537–1538.

22. Feller M, Huwiler K, Stephan R et al. Mycobacterium avium subspecies paratuberculosis and Crohn’s disease: a systematic review and meta-analysis. Lancet Infect Dis 2007; 7: 607–613.

23. Ryan P, Bennett MW, Aarons S et al. PCR detection of Mycobacterium paratuberculosis in Crohn’s disease granulomas isolated by laser capture microdissection. Gut 2002; 51: 665–670.

24. Ryan P, Kelly RG, Lee G et al. Bacterial DNA within granulomas of patients with Crohn’s disease-detection by laser capture microdissection and PCR. Am J Gastroenterol 2004; 99(8): 1539–1543.

25. Naser SA, Ghobrial G, Romero C et al. Culture of Mycobacterium avium subspecies paratuberculosis from the blood of patients with Crohn’s disease. Lancet 2004;364(9439): 1039–1044.

26. Hermon-Taylor J, Barnes N, Clarke C et al. Mycobacterium paratuberculosis cervical lymphadenitis, followed five years later by terminal ileitis similar to Crohn’s disease. BMJ 1998; 316(7129): 449–453.

27. Marks DJ, Harbord MW, MacAllister R et al. Defective acute inflammation in Crohn’s disease: a clinical investigation. Lancet 2006; 367(9511): 668–678.

28. Simms LA, Doecke JD, Walsh MD et al. Reduced alpha-defensin expression is associated with inflammation and not NOD2 mutation status in ileal Crohn’s disease. Gut 2008; 57: 903–910.

29. Ogura Y, Lala S, Xin W et al. Expression of NOD2 in Paneth cells: a possible link to Crohn’s ileitis. Gut 2003; 52(11): 1591–1597.

30. Stabel JR. Transitions in immune responses to Mycobacterium paratuberculosis. Vet Microbiol 2000; 77(3–4): 465–473.

31. Sechi LA, Gazouli M, Ikonomopoulos J et al. Mycobacterium avium subsp. paratuberculosis, genetic susceptibility to Crohn’s disease, and Sardinians: the way ahead. J Clin Microbiol 2005; 43(10): 5275–5277.

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

33. Wang G, Stange EF, Wehkamp J. Host-microbe interaction: mechanisms of defensin deficiency in Crohn’s disease. Expert Rev Anti Infect Ther 2007; 5(6): 1049–1057.

34. Wehkamp J, Wang G, Kübler I et al. The Paneth cell alpha-defensin deficiency of ileal Crohn’s disease is linked to Wnt/Tcf-4. J Immunol 2007; 179(5): 3109–3118.

35. Fixa B, Komárková O, Bedrna J et al. Crohnova choroba a protilátky proti Mycobacterium avium subspecies paratuberculosis. Česká a slovenská gastroenterologie 2000; 54(2): 175–179.

36. Hermon-Taylor J. Mycobacterium avium subspecies paratuberculosis in the causation of Crohn’s disease. World J Gastroenterol 2000; 6(5): 630–632.

37. Fixa B, Komárková O, Skaunic V et al. Inhibition of leucocyte migration by antigens from human colon and E. coli O 14 in patients with ulcerative colitis. Scand J Gastroenterol, 1975; 10: 491–493.

38. Rowbotham DS, Howdle PD, Trejdosiewicz LK. Peripheral cell-mediated immune response to mycobacterial antigens in inflammatory bowel disease. Clin Exp Immunol 1995;102(3): 456–461.

39. Ren Z, Turton J, Borody T et al. Selective Th2 pattern of cytokine secretion in Mycobacterium avium subsp. paratuberculosis infected Crohn’s disease. J Gastroenterol Hepatol 2008; 23(2): 310–314.

40. Jones PH, Farver TB, Beaman B et al. Crohn’s disease in people exposed to clinical cases of bovine paratuberculosis. Epidemiol Infect 2006; 134(1): 49–56.

41. Sartor RB. Does Mycobacterium avium subspecies paratuberculosis cause Crohn’s disease? Gut 2005; 54(7): 896–898.

42. Greenstein RJ, Su L, Haroutunian V et al. On the Action of Methotrexate and 6-Mercaptopurine on M. avium subspecies paratuberculosis. PLoS One 2007; 2(1): e161.

43. Greenstein RJ, Su L, Shahidi A et al. On the Action of 5-Amino-Salicylic Acid and Sulfapyridine on M. avium including Subspecies paratuberculosis. PLoS One 2007; 2(6): e516.

44. Mendoza JL, Lana R, Díaz-Rubio M. Mycobacterium avium subspecies paratuberculosis and its relationship with Crohn’s disease. World J Gastroenterol 2009; 15(4): 417–422.

45. Graham DY, Al-Assi MT, Robinson M. Prolonged remission in Crohn’s disease following therapy for Mycobaterium paratuberculosis. Gastroenterol 1995; 108: A826.

46. Gui GP, Thomas PR, Tizard ML et al. Two-year-outcomes analysis of Crohn’s disease treated with rifabutin and macrolide antibiotics. J Antimicrob Chemother 1997; 39: 393–400.

47. Borgaonkar MR, MacIntosh DG, Fardy JM. A meta-analysis of antimycobacterial therapy for Crohn’s disease. Am J Gastroenterol 2000; 95: 725–729.

48. Borody TJ, Bilkey S, Wettstein AR et al. Anti-mycobacterial therapy in Crohn’s disease heals mucosa with longitudinal scars. Dig Liver Dis 2007; 39(5): 438–444.

49. Selby W, Pavli P, Rotty B et al. Two-Year Combination Antibiotic Therapy With Clarithromycin, Rifabutin, and Clofazimine for Crohn’s Disease. Gastroenterol 2007; 132(7): 2313–2319.

50. Hruska K, Bartos M, Kralik P et al. Mycobacterium avium subsp. paratuberculosis in powdered infant milk: paratuberculosis in cattle – the public health problem to be solved. Vet Med-Czech 2005; 50: 327–335.

51. Naser S.A, Schwartz D., Shafran I. Isolation of Mycobacterium avium subsp paratuberculosis from breast milk of Crohn’s disease patients. AJG 2000; 95(4): 1094–1095.

Autoři děkují za významnou pomoc při získávání a ukládání vzorků ze střevních resekátů paní Ireně Šulcové z 2. interní katedry LF UK v Hradci Králové.

Paediatric gastroenterology Gastroenterology and hepatology Surgery

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