The effect of oxygen on endotoxin production in bacteria of the Bacteroides fragilis group isolated from patients with colorectal carcinoma
Authors:
D. Chmelař 1,2; M. Hájek 1,3; J. Janečková 4; J. Vobejdová 4; P. Martineková 1; A. Kašíková 1
Authors‘ workplace:
Katedra biomedicínských oborů, Ústav mikrobiologie a imunologie, Lékařská fakulta Ostravské univerzity v Ostravě
1; Referenční laboratoř ČR pro anaerobní bakterie, Lékařská fakulta Ostravské univerzity v Ostravě
2; Centrum hyperbarické medicíny, Městská nemocnice Ostrava
3; Oddělení infekční diagnostiky, Litomyšlská nemocnice, Nemocnice Pardubického kraje
4
Published in:
Epidemiol. Mikrobiol. Imunol. 65, 2016, č. 2, s. 129-135
Category:
Original Papers
Overview
Objective:
The aim of the study was to draw attention to the risk posed by anaerobic bacteria of the Bacteroides fragilis (BAFR) group, isolated particularly from abdominal lesions, and to assess the possible role of these species in colorectal cancer. A correlation has previously been suggested between the detection of the bacteria of the genus Bacteroides in patients on a meat-based diet and intestinal and, in particular, colorectal cancer. Given that the species of the BAFR group are major producers of endotoxins, measurements and statistical analysis of endotoxin production were used to compare the Bacteroides strains isolated from clinical specimens of patients with colon cancer, rectal cancer, and other abdominal lesions.
Materials and Methods:
Endotoxin production was detected in bacterial strains of the BAFR group (B. fragilis, B. thetaiotaomicron, B. distasonis, and B. vulgatus) isolated from clinical specimens of patients with rectal cancer, colon cancer, and intestinal cancer and was compared with that in strains from samples of patients with inflammatory conditions (anal abscess, appendicitis, skin abscess, etc.) under anaerobic and microaerophilic (with 5% of oxygen) culture conditions. The production of endotoxins was detected quantitatively using the Pyrosate LAL assay kit (Limulus Amoebocyte Lysate Test, BIOGENIX, CR) in four species of the BAFR group after anaerobic and microaerophilic culture. Five strains of each isolated Bacteroides species from each type of specimens were tested (in total 140 BAFR strains). The amount of endotoxin was given in endotoxin units per ml (EU/ml).
Results:
Endotoxin production by bacteria under microaerophilic culture conditions was several times higher in comparison with strictly anaerobic culture.
The difference was statistically significant (F1.269 = 160, p <0.0001). As regards the effect of oxygen on endotoxin production, the amount of endotoxins produced under microaerophilic culture conditions (average 889.1 EU/ml) was 2.5 times as high as that observed under anaerobic culture conditions (358.2 EU/ml), regardless of the bacteroides species and diagnosis. These results suggest that the amount of free oxygen in the environment affects the amount of endotoxin generated by the Bacteroides strains.
Conclusion:
The results show that endotoxin production by the Bacteroides strains under microaerophilic culture conditions is several times as high as that under strictly anaerobic culture conditions.
KEYWORDS:
Bacteroides fragilis group – endotoxins – colorectal carcinoma
Sources
1. Bang FA. Bacterial Disease of Limulus polyphemus. Bull J Hopkins Hosp, 1956;98:325–330.
2. Čižnár I. Toxíny gramnegatívnych črevných baktérií. Bratislava, VEDA, 1982, 15–69.
3. Crawley, MJ Statistical Computing. An Introduction to data analysis using S-Plus. Chichester, John Wiley and Sons, 2002.
4. Delahooke, DM, Barclay GR, Poxton IR. Tumor necrosis factor induction by an aqueous phenol-extracted lipopolysacchyride complex from Bacteroides species. Infect. Immun., 1995,63: 840–846.
5. Fukata M, Abreu MT. Role of Toll-like receptors in gastrointestinal malignancies. Oncogene, 2008;27(2):234–243.
6. Gee JM, Rocha ER, Smith CJ. Examination of the Contributions of DPS to the Oxidative Stress response and Pathogenesis of the Oportunistic Pathogen, Bacteroides fragilis. The 9th Biennial Congress of ther Anaerobe Society of Americas, 2008.
7. Goldner M, Mingot M, Emond JP, Dublanchet A. Influence of different levels of redox potential on fermentative products formed by Bacteroides fragilis. Clin Ifect Dis, 1997;25 (Supl.):S147–S150.
8. Chmelař D. Gastrointestinální trakt – zdroj endogenních anaerobních infekcí. Správy klinickej mikrobiológie, 2001, (Supl. A):87–88. ISSN 1335-8219.
9. Chmelař D, Vrtný J. Produkce endotoxinu u bakterií Bacteroides fragilis group ve vztahu k onemocnění karcinomem tlustého střeva a rekta u lidí. Klin Mikrobiol Inf Lek, 2010;16(3):97–102.
10. Joiner KA, McAdam KP, Kasper DL. Lipopolysaccharides from Bacteroides fragilis are mitogenic for spleen cells from endotoxin responder and nonresponder mice. Infec. Immun, 1982;36(3):1139–1145.
11. Kozelská R. Průkaz endotoxinů u patogenních gramnegativních bakterií. Bakalářská práce, Katedra vyšetřovacích metod a lékařské biologie, Zdravotně sociální fakulta, Ostravská univerzita, 2006:29–32.
12. Krásová Z, Matušek A, Chmelař D. Hyperbaric oxygenation in the treatment of necrotizing fasciitis. Vnit. Lek, 1992;36 (7):640–644.
13. Lewis CJ, Coby BA. Oxidant production by APCs in response to glycoantigens. The Journal of Immunology, 2007;178:36–39.
14. Lien E, Ingalls RR. Toll-like receptors. Critical Care Medicine, 2002;30(1), Suppl:1–11.
15. Matušek A, Martínek L, Chmelař D, Krásová Z. Fournierova gangréna. Rozhledy v chirurgii, 1998;1:24–26.
16. Morris JG. Characteristics of anaerobic metabolism. In: Duerden, B. I, Drasar, B. S (eds). Anaerobes in Human Disease., London Edward Arnold, 1991:16–37.
17. Muta T, Takeshige K. Essential roles of CD14 and lipopolysaccharide-binding protein for activation of toll-like receptor (TLR)2 as well as TLR4 Reconstitution of TLR2- and TLR4-activation by distinguishable ligands in LPS preparations. Eur J Biochem, 2001;268(16): 4580–4589.
18. Namavar F, Theunissen EB, Verweij-Van Vught AMJJ. Epidemiology of the Bacteroides fragilis group in the colonic flora of patients with colonic cancer. J Med Microbiol, 1989;29: 171–176.
19. Petruželka L, Konopásek B. Klinická onkologie. Praha. Karolinum, 2003, ISBN 80-246-0395-0.
20. R Development Core Team. A language and environment for statistical computing. R Foundation for Statistical Computing, 2004, Vienna, Austria. Dostupné na www: < http://www.R-project.org>
21. Reott MA, Rocha ER, Parker AC, Smith CJ. Thioredoxins in Redox Maintenance and Survival During Oxidative Stress of Bacteroides fragilis. The 9th Biennial Congress of the Anaerobe Society of Americas, 2008.
22. Rocha ER, Selby T, Coleman JP, Smith CJ. Oxidative stress response in an anaerobe, Bacteroides fragilis: a role for catalase in protection against hydrogen peroxide. Journal of Bacteriology, 1996;178(23):6895–6903.
23. Rocha ER, Smith CJ. Role of the alkyl hydroperoxide reductase (ahpCF) gene in oxidative stress defense of the obligate Anaerobe Bacteroides fragilis. Journal of Bacteriology, 1999;181(18):5701–5710.
24. Rokosz A, Kruszewska D, Rouyan GS, Meisel-Mikołajczyk F. Detection of endotoxins and enterotoxins of Bacteroides fragilis in culture media. Med Dosw Mikrobiol, 1997; 49 (1–2):61–67.
25. Rokosz A, Meisel-Mikołajczyk F, Malchar C, Nowaczyk M, Górski A. Stimulation of adhesion molecules on vascular endothelium by capsular polysaccharide, lipopolysaccharide and components of lipopolysaccharide from Bacteroides thetaiotaomicron. Med Dosw Mikrobiol, 2000;52(3):275–282.
26. Sears CL. Enterotoxigenic Bacteroides fragilis:a rogue among symbiotes. Clin Microbiol Rev, 2009;22(2):349–369.
27. Sears CL, Geis AL, Housseau F. Bacteroides fragilis subverts mucosal biology: from symbiont to colon carcinogenesis. J Clin Invest, 2014;124(10): 4166–4172.
28. Shear M. Chemical Treatment of Tumors IX: reactions of mice with primary subcutaneous tumors. J Natl Cancer Inst, 1944; 4:461–476.
29. Smith CJ. The Bacteroides fragilis Oxidative Stress Response During Intra-Abdominal Infections. The 9th Biennial Congress of ther Anaerobe Society of Americas, 2008.
30. Tang YP, Dallas MM, Malamy MH. Characterization of the Bat I (Bacteroides aerotolerance) operon in Bacteroides fragilis isolation of a B. fragilis mutant with reduced aerotolerance and impaired growth in in vivo model systems. Mol Microbiol, 1999;32:139–149.
31. Vangelakos CG, Rhee K-J, Wu S, Gan C, Sears CL. Bacteroides fragilis and Other Bacteroides Species Demonstrate Marked Aerotolerance. The 9th Biennial Congress of ther Anaerobe Society of Americas, 2008.
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Hygiene and epidemiology Medical virology Clinical microbiologyArticle was published in
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