Original Article
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World J Gastroenterol. Feb 7, 2011; 17(5): 609-617
Published online Feb 7, 2011. doi: 10.3748/wjg.v17.i5.609
Bacteriocinogeny in experimental pigs treated with indomethacin and Escherichia coli Nissle
Jan Bures, David Smajs, Jaroslav Kvetina, Miroslav Förstl, Jan Smarda, Darina Kohoutova, Martin Kunes, Jiri Cyrany, Ilja Tacheci, Stanislav Rejchrt, Jirina Lesna, Viktor Vorisek, Marcela Kopacova
Jan Bures, Darina Kohoutova, Jiri Cyrany, Ilja Tacheci, Stanislav Rejchrt, Marcela Kopacova, 2nd Department of Medicine, Charles University in Praha, Faculty of Medicine at Hradec Kralove, University Teaching Hospital, 500 05 Hradec Kralove, Czech Republic
David Smajs, Jan Smarda, Department of Biology, Masaryk University, Faculty of Medicine, 625 00 Brno, Czech Republic
Jaroslav Kvetina, Martin Kunes, Institute of Experimental Biopharmaceutics, Joint Research Centre of Czech Academy of Sciences and PRO.MED.CS Praha a.s., 500 02 Hradec Kralove, Czech Republic
Miroslav Förstl, Jirina Lesna, Institute of Clinical Microbiology, Charles University in Praha, Faculty of Medicine at Hradec Kralove, University Teaching Hospital, 500 05 Hradec Kralove, Czech Republic
Viktor Vorisek, Institute of Clinical Biochemistry and Diagnostics, Charles University in Praha, Faculty of Medicine at Hradec Kralove, University Teaching Hospital, 500 05 Hradec Kralove, Czech Republic
Author contributions: All authors contributed equally to this work.
Supported by Research project MZO 00179906 from the Ministry of Health of the Czech Republic, by institutional support from the Czech Republic (MSM0021622415) and by research grants GAČR 305/080535 and NS9665-4/2008 (Ministry of Health of the Czech Republic)
Correspondence to: Jan Bures, MD, PhD, Professor, 2nd Department of Medicine, Charles University in Praha, Faculty of Medicine at Hradec Kralove, University Teaching Hospital, 500 05 Hradec Kralove, Czech Republic. bures@lfhk.cuni.cz
Telephone: +420-495-834240 Fax: +420-495-834785
Received: August 3, 2010
Revised: September 25, 2010
Accepted: October 2, 2010
Published online: February 7, 2011
Abstract

AIM: To evaluate bacteriocinogeny in short-term high-dose indomethacin administration with or without probiotic Escherichia coli Nissle 1917 (EcN) in experimental pigs.

METHODS: Twenty-four pigs entered the study: Group A (controls), Group B (probiotics alone), Group C (indomethacin alone) and Group D (probiotics and indomethacin). EcN (3.5 × 1010 bacteria/d for 14 d) and/or indomethacin (15 mg/kg per day for 10 d) were administrated orally. Anal smears before and smears from the small and large intestine were taken from all animals. Bacteriocin production was determined with 6 different indicator strains; all strains were polymerase chain reaction tested for the presence of 29 individual bacteriocin-encoding determinants.

RESULTS: The general microbiota profile was rather uniform in all animals but there was a broad diversity in coliform bacteria (parallel genotypes A, B1, B2 and D found). In total, 637 bacterial strains were tested, mostly Escherichia coli (E. coli). There was a higher incidence of non-E. coli strains among samples taken from the jejunum and ileum compared to that of the colon and rectum indicating predominance of E. coli strains in the large intestine. Bacteriocinogeny was found in 24/77 (31%) before and in 155/560 (28%) isolated bacteria at the end of the study. Altogether, 13 individual bacteriocin types (out of 29 tested) were identified among investigated strains. Incidence of four E. coli genotypes was equally distributed in all groups of E. coli strains, with majority of genotype A (ranging from 81% to 88%). The following types of bacteriocins were most commonly revealed: colicins Ia/Ib (44%), microcin V (18%), colicin E1 (16%) and microcin H47 (6%). There was a difference in bacteriocinogeny between control group A (52/149, 35%) and groups with treatment at the end of the study: B: 31/122 (25%, P = 0.120); C: 43/155 (28%, P = 0.222); D: 29/134 (22%, P = 0.020). There was a significantly lower prevalence of colicin Ib, microcins H47 and V (probiotics group, P < 0.001), colicin E1 and microcin H47 (indomethacin group, P < 0.001) and microcins H47 and V (probiotics and indomethacin group, P = 0.025) compared to controls. Escherichia fergusonii (E. fergusonii) was identified in 6 animals (6/11 isolates from the rectum). One strain was non-colicinogenic, while all other strains of E. fergusonii solely produced colicin E1. All animals started and remained methanogenic despite the fact that EcN is a substantial hydrogen producer. There was an increase in breath methane (after the treatment) in 5/6 pigs from the indomethacin group (C).

CONCLUSION: EcN did not exert long-term liveability in the porcine intestine. All experimental pigs remained methanogenic. Indomethacin and EcN administered together might produce the worst impact on bacteriocinogeny.

Keywords: Bacteriocinogeny, Escherichia coli Nissle 1917, Experimental pigs, Indomethacin