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Clostridium difficile is the etiological agent of diarrhoea and colitis, especially in elderly patients. The incidence of these diseases has increased during the last 10 years. Emergence of so-called hypervirulent strains is considered as one of the main factors responsible for the more severe disease and changed profile of sensitivity to antimicrobial agents. The aim of this work was to determine the sensitivity profile of toxigenic strains of C. difficile in the Czech Republic in 2011-2012 to selected antibiotics. The antibiotics clindamycin, metronidazole, vancomycin and amoxicillin with clavulanic acid were used for this purpose. Isolates cultured on Brazier's C. difficile selective agar were analysed for the presence of toxin genes using Xpert detection system. Xpert analysis revealed that 33 strains carried the genes for toxins tcdB, cdt and tcdCΔ117, thus showing characteristics typical for the hypervirulent ribotype 027/PFGE type NAP1/REA type B1. The remaining 29 strains carried only the gene for toxin B (tcdB) and not cdt and tcdCΔ117. Our results indicate the higher susceptibility of C. difficile hypertoxigenic strains to three out of four tested antibiotics (except vancomycin) than it is for the other toxigenic strains. We found that only 10.34% of other toxigenic strains were resistant to clindamycin, and no resistance was found in all other cases. All the isolates were sensitive to amoxicillin/clavulanic acid in vitro. However, its use is not recommended for therapy of infections caused by C. difficile.

Článek

Enumeration of Mycobacterium avium subsp. paratuberculosis by quantitative real-time PCR, culture on solid media and optical densitometry

BMC research notes. 2012 ; 5 () : 114.

BMC Res Notes
ISSN 1756-0500
Medvik
Zdroj

BACKGROUND: Different approaches are used for determining the number of Mycobacterium avium subsp. paratuberculosis (MAP) cells in a suspension. The majority of them are based upon culture (determination of CFU) or visual/instrumental direct counting of MAP cells. In this study, we have compared the culture method with a previously published F57 based quantitative real-time PCR (F57qPCR) method, to determine their relative abilities to count the number of three different MAP isolates in suspensions with the same optical densities (OD). McFarland turbidity standards were also compared with F57qPCR and culture, due to its frequent inclusion and use in MAP studies. FINDINGS: The numbers of MAP in two-fold serial dilutions of isolates with respective OD measurements were determined by F57qPCR and culture. It was found that culture provided lower MAP CFU counts by approximately two log10, compared to F57qPCR. The McFarland standards (as defined for E. coli) showed an almost perfect fit with the enumeration of MAP performed by F57qPCR. CONCLUSIONS: It is recommended to use culture and/or qPCR estimations of MAP numbers in experiments where all subsequent counts are performed using the same method. It is certainly not recommended the use of culture as the standard for qPCR experiments and vice versa.

MeSH
bakteriální nálož metody MeSH
denzitometrie MeSH
DNA bakterií analýza MeSH
Escherichia coli cytologie MeSH
feces mikrobiologie MeSH
kultivační média MeSH
kvantitativní polymerázová řetězová reakce MeSH
Mycobacterium avium subsp. paratuberculosis cytologie genetika izolace a purifikace MeSH
nemoci skotu diagnóza mikrobiologie MeSH
paratuberkulóza diagnóza mikrobiologie MeSH
skot MeSH
zvířata MeSH
Check Tag
skot MeSH
zvířata MeSH
Publikační typ
časopisecké články MeSH
práce podpořená grantem MeSH

Článek

Comparison of the conventional culture, the manual fluorescent MGIT system and the automated fluorescent MGIT 960 culture system for the detection of Mycobacterium avium ssp. avium in tissues of naturally infected hens

Folia microbiologica. 2009 ; 54 (2) : 137-141.

ISSN 0015-5632
Medvik
Zdroj

Different methods for the detection of Mycobacterium avium ssp. avium (MAA) in naturally infected hens were compared. They included the conventional culture method (solid Herrold's and Stonebrink media and liquid Sula medium) and newly developed liquid culture systems, the manual mycobacteria growth indicator tube (M-MGIT) and the fully automated BACTEC MGIT 960 system (A-MGIT). 152 tissues originating from 15 naturally infected hens have been processed. The overall detection rates (percentage of positive cultures from the number of positive cultures determined by all the methods together) were 60, 70 and 76 % for the conventional media, M-MGIT and A-MGIT systems, respectively, the mean time of mycobacteria detection being 32.6, 17.6 and 14.6 d, respectively. The lowest contamination rate (2.0 %) was found in A-MGIT compared with M-MGIT (4.6 %) and conventional media (10.4 %).

Časopis

Ostravské dny hyperbarické medicíny : sborník přednášek
[Mezinárodní workshop potápěčské medicíny]

Ostravské dny hyperbarické medicíny (konference) (2. : 2010 : Velké Karlovice, Česko)
Autor Autorita
Hájek, Michal, 1969-
Autor Autorita ORCID
Novomeský, František, 1947-
Autor Autorita
Beran, Vladimír, 1979-
Autor Autorita
Pudil, Radek, 1966-
Autor Autorita ORCID
Klugar, Miloslav, 1982-
Autor Autorita ORCID
Chmelař, Dittmar, 1955-
Autor Autorita
Městská nemocnice (Ostrava, Česko). Centrum hyperbarické medicíny
Autor Autorita
Česká společnost hyperbarické a letecké medicíny ČLS JEP
Autor Autorita
Ostravská univerzita. Lékařská fakulta
Autor Autorita

V Ostravě : Ostravská univerzita, Fakulta zdravotnických studií, 2008- | 2012- Ostrava : Ostravská univerzita v Ostravě, Lékařská fakulta

svazků : ilustrace ; 21 cm

Konspekt
Fyzioterapie. Psychoterapie. Alternativní lékařství
NLK Obory
hyperbarická a letecká medicína

Článek

Distribution of mycobacteria in clinically healthy ornamental fish and their aquarium environment

Journal of fish diseases. 2006 ; 29 (7) : 383-393.

Medvik
Zdroj

Some mycobacterial species (particularly Mycobacterium marinum) found in aquarium environments may cause chronic diseases in fish and cutaneous infections in humans, the so-called 'fish tank granuloma'. The presence and distribution of mycobacterial species in clinically healthy aquarium fish and their environment has not been adequately explored. The present study analysed the occurrence of mycobacteria in a decorative aquarium (Brno, South Moravia) and in five aquaria of a professional fish breeder (Bohumin, North Moravia). After Ziehl-Neelsen staining, acid-fast rods (AFR) were observed in six (14.3%) and mycobacteria were detected by culture in 18 (42.9%) of 42 tissue samples from 19 fish. Sixty-five samples of the aqueous environment from all six aquaria were examined; AFR were found in 16 (24.6%) and mycobacteria were detected by culture in 49 (75.4%) samples. Forty-one (70.7%) of 58 selected mycobacterial isolates were identified biochemically as follows: M. fortuitum, M. flavescens, M. chelonae, M. gordonae, M. terrae, M. triviale, M. diernhoferi, M. celatum, M. kansasii and M. intracellulare. The clinically important species for humans and fish, M. marinum, was not detected. Mycobacterium kansasii was isolated from one sample of the aquarium environment from North Moravia, which is a region of the Czech Republic with endemic incidence of M. kansasii in water. The incidence of other conditionally pathogenic mycobacterial species in healthy fish and in all investigated constituents of the aquarium environment including snails and crustaceans used for fish feeding, was quite high. Accordingly, mycobacterial species from aquarium environments may serve as a possible source of infection for both aquarium fish and immunodeficient fish handlers.

MeSH
atypické mykobakteriální infekce epidemiologie veterinární MeSH
ekosystém MeSH
financování organizované MeSH
incidence MeSH
nemoci ryb epidemiologie mikrobiologie MeSH
netuberkulózní mykobakterie izolace a purifikace klasifikace MeSH
rybářství MeSH
ryby MeSH
životní prostředí MeSH
zvířata MeSH
Check Tag
zvířata MeSH
Publikační typ
srovnávací studie MeSH

Článek

Identification of members of Mycobacterium avium species by Accu-Probes, serotyping, and single IS900, IS901, IS1245 and IS901-flanking region PCR with internal standards

Journal of microbiological methods. 2006 ; 64 (3) : 333-345.

ISSN 0167-7012
Medvik
Zdroj

From Mycobacterium avium species Mycobacterium avium subsp. paratuberculosis (n=961), Mycobacterium a. avium (n=677), Mycobacterium a. silvaticum (n=5), and Mycobacterium a. hominissuis (n=1566) were examined, and from Mycobacterium tuberculosis complex M. tuberculosis (n=2), Mycobacterium bovis (n=13), M. bovis BCG (n=4), and Mycobacterium caprae (n=10) were examined. From other mycobacterial species Mycobacterium intracellulare (n=60) and atypical mycobacteria (n=256) including Mycobacterium fortuitum, Mycobacterium chelonae, Mycobacterium scrofulaceum, Mycobacterium gastri and other species of conditionally pathogenic mycobacteria were analysed. The internal standard molecules corresponding to insertion sequences IS900, IS901, IS1245, and flanking region (FR300) of IS901 were produced by PCR of alfalfa genome segment and inserted into plasmid vector. The resulting recombinant plasmid molecules were used as internal standards in coamplification with a total of 4729 mycobacterial collection strains and field isolates between 1996 and 2003. The size differences between amplicons obtained from IS900 (258 bp), IS901 (1108 bp), IS1245 (427 bp), and FR300 (300 bp) and from corresponding internal standard molecules ISIS900 (591 bp), ISIS901 (1 336 bp), ISIS1245 (583 bp), and IS901 flanking region of 300 bp ISFR300 (488 bp), respectively, allowed easy discrimination. The internal amplicons were visible by naked aye on agarose gel when 10(1), 10(3), 10(2), and 10(2) molecules for ISIS900, ISIS901, ISIS1245, and ISFR300 were used in the PCR, respectively, when no bacterial DNA was added to the reaction. The system was tested to define the amount of internal standards that could be used in the PCR without affecting the amplification of the specific segment. Non-specific amplifications were observed in M. fortuitum with IS1245 PCR and mixed infections with M. a. avium and M. a. hominissuis from pigs and cattle were found. PCR results of typing were compared with serotyping and Accu-Probes analyses in selected field isolates.