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BackgroundColorectal cancer (CRC) is the 3rd most common cancer worldwide and the Czech Republic has the 6th highest incidence of CRC worldwide. Large intestinal microbiota play in its etiopathogenesis important role. Bacteriocins are proteins, produced by bacteria from the Enterobacteriaceae family. The aim of our prospective study was to assess the colonization of large intestinal mucosa by Escherichia coli strains and to investigate their bacteriocin production.MethodsA total of 30 consecutive patients with colorectal adenoma, CRA (17 men, 13 women, aged 39–79, mean age 63 ± 9), 30 patients with CRC (23 men, 7 women, aged 38–86, mean age 67 ± 11) and 20 healthy controls (9 men, 11 women, age 23–84, mean age 55 ± 15) were enrolled into prospective study. Mucosal biopsies were taken in the caecum, transverse colon and rectum during pancolonoscopy. Microbiological culture, isolation and identification of bacteria followed. Bacteriocin production was assessed by growth inhibition of indicator strains E. coli K12-Row, E. coli C6 (phi), and Shigella sonnei 17. Identification of bacteriocin-encoding determinants and E. coli phylogroups was performed using PCR methods.ResultsA total of 622 strains were isolated and further investigated. A significantly higher frequency of simultaneous production of colicins and microcins was revealed in the group of patients with CRC, when compared to patients with CRA, p = 0.031. A significantly higher frequency of E. coli phylogroup D was found in patients with CRC, when compared to controls, p = 0.044. A significantly higher prevalence of bacteriocinogeny was confirmed in patients with advanced adenoma when compared to patients with non-advanced adenoma, p = 0.010. Increasing bacteriocinogeny was associated with an increasing stage of CRC (assessed according to TNM classification). Either E. coli phylogroup B2 or E. coli phylogroup D were isolated in biopsies of patients with right-sided CRC. A statistically higher incidence of E. coli phylogroup B2 was found in patients with right-sided CRC when compared to patients with left-sided CRC, p = 0.028.ConclusionsLarge intestinal mucosa of patients with more advanced colorectal neoplasia is colonized with more virulent strains of E. coli and higher production of bacteriocins is observed in these patients when compared to those with less advanced colorectal neoplasia.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-014-0733-7) contains supplementary material, which is available to authorized users.

Mycobacteria cause serious infections in animals and human beings. Huge economic losses on farms are caused by selected species of this wide family. A high risk of transmission of infection from animal to human exists. The knowledge of exact pathogen characteristics is an important factor which can improve quick and adequate healing. Cultivation and determination of phenotype is still the "gold standard", but has the disadvantage of taking a long time and also low detection limit. Biochemical characterisation of isolates is not exact, and it is expensive. A more popular method used is the amplification of specific loci by polymerase chain reaction (PCR). For this method, the isolation of sufficient amounts of purified DNA is necessary. In this paper the most frequently used method for DNA isolation from live mycobacterial cells, body fluids, tissues, histological samples and forensic materials are outlined. This paper assists only as guide for these methods, so we describe them briefly.Keywords: Johne's disease; Crohn's disease; zoonoses List of abbreviations: BCG = Bacille Calmette-Guérin; CB18 = C18-carboxypropylbetain; CFU = colony forming units; CPH = N-cetylpyridinium chloride, cerebrospinal fluid; CTAB = cetyltrimethyl ammonium bromide; DNA = deoxyribonucleic acid; DR = direct repeat domain; DTT = dithiotreitol; EDTA = disodium ethylene diamine tetraacetate 2 H 2 O; FB = freezing and boiling; HIV/AIDS = Human Immune-Deficiency Virus/Acquired Immune Deficiency Syndrome; IgG = class G immunoglobulin; IMS = immunomagnetic separation; IS = insertion sequence; LCM = laser capture micro-dissection; MAC = Mycobacterium avium complex; MOTT = mycobacteria other than tuberculosis; MTC = Mycobacterium tuberculosis complex; NALC = N-acetyl-L-cystein; OTN-PCR = one-tube nested polymerase chain reaction; PBS = phosphate-buffered saline; PCI = phenol-chloroform-isoamylalcohol; PCR = polymerase chain reaction; PRA-PCR = polymerase restriction analysis-polymerase chain reaction; RFLP = restriction fragment length polymorphism; rRNA = ribosomal ribonucleic acid; SDS = sodium dodecyl sulphate; TE = Tris-HCl + EDTA buffer; TLC = thin-layer chromatography; TSA = tuberculostearic acid: 10-methyloktadekan acid

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ABSTRACT:Interstitial pneumonia (2/3 of the lungs were affected) and diffusely enlarged bronchial and mediastinal lymph nodes were diagnosed by gross examination of a dead 16-year-old mare. Based on histopathological examination and the detection of acid-fast rods after staining by the Ziehl-Neelsen technique, tuberculosis was suspected. Mycobacterium avium subsp. avium of serotype 2 and IS901+/IS1245+ genotype was isolated from the pulmonary lymphnode after five-week incubation at 37°C. Due to the fact that horses have a naturally high resistance to mycobacterial infections, the high age of the mare most likely contributed to the development of the disease.