This preliminary study focused on the effect of exposure to 0.5 T static magnetic fields on Escherichia coli adhesion and orientation. We investigated the difference in bacterial adhesion on the surface of glass and indium tin oxide-coated glass when exposed to a magnetic field either perpendicular or parallel to the adhesion surface (vectors of magnetic induction are perpendicular or parallel to the adhesion surface, respectively). Control cultures were simultaneously grown under identical conditions but without exposure to the magnetic field. We observed a decrease in cell adhesion after exposure to the magnetic field. Orientation of bacteria cells was affected after exposure to a parallel magnetic field. On the other hand, no effect on the orientation of bacteria cells was observed after exposure to a perpendicular magnetic field.
- MeSH
- bakteriální adheze * MeSH
- Escherichia coli fyziologie MeSH
- magnetické pole * MeSH
- sklo * MeSH
- sloučeniny cínu * MeSH
- Publikační typ
- časopisecké články MeSH
The aim of this study was to determine possible differences in the faecal microbiota of dairy cows infected with Mycobacterium avium subsp. paratuberculosis (Johne's disease) in comparison with noninfected cows from the same herds. Faecal samples from cows in 4 herds were tested for M. avium subsp. paratuberculosis by real-time PCR, and faecal bacterial populations were analysed by 454 pyrosequencing of the 16S rRNA gene. The most notable differences between shedding and nonshedding cows were an increase in the genus Psychrobacter and a decrease in the genera Oscillospira, Ruminococcus, and Bifidobacterium in cows infected with M. avium subsp. paratuberculosis. The present study is the first to report the faecal microbial composition in dairy cows infected with M. avium subsp. paratuberculosis.
- MeSH
- feces mikrobiologie MeSH
- mlékárenství MeSH
- Mycobacterium avium subsp. paratuberculosis genetika izolace a purifikace fyziologie MeSH
- nemoci skotu mikrobiologie MeSH
- paratuberkulóza mikrobiologie MeSH
- sekvenční analýza DNA veterinární MeSH
- skot mikrobiologie MeSH
- vylučování bakterií z těla MeSH
- zvířata MeSH
- Check Tag
- skot mikrobiologie MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
A high intracellular concentration of potassium (200-300 mmol/L) is essential for many yeast cell functions, such as the regulation of cell volume and pH, maintenance of membrane potential, and enzyme activation. Thus, cells use high-affinity specific transporters and expend a lot of energy to acquire the necessary amount of potassium from their environment. In Candida genomes, genes encoding 3 types of putative potassium uptake systems were identified: Trk uniporters, Hak symporters, and Acu ATPases. Tests of the tolerance and sensitivity of C. albicans, C. dubliniensis, C. glabrata, C. krusei, C. parapsilosis, and C. tropicalis to various concentrations of potassium showed significant differences among the species, and these differences were partly dependent on external pH. The species most tolerant to potassium-limiting conditions were C. albicans and C. krusei, while C. parapsilosis tolerated the highest KCl concentrations. Also, the morphology of cells changed with the amount of potassium available, with C. krusei and C. tropicalis being the most influenced. Taken together, our results confirm potassium uptake and accumulation as important factors for Candida cell growth and suggest that the sole (and thus probably indispensable) Trk1 potassium uptake system in C. krusei and C. glabrata may serve as a target for the development of new antifungal drugs.
Gene expression profiles are important data to reveal the functions of genes putatively involved in crucial biological processes. RNA arbitrarily primed polymerase chain reaction (RAP-PCR) and specifically primed reverse transcription polymerase chain reaction (RT-PCR) were combined to screen differentially expressed genes following development of a commercial Bacillus thuringiensis subsp. kurstaki strain 8010 (serotype 3a3b). Six differentially expressed transcripts (RAP1 to RAP6) were obtained. RAP1 encoded a putative triple helix repeat-containing collagen or an exosporium protein H related to spore pathogenicity. RAP2 was homologous to a ClpX protease and an ATP-dependent protease La (LonB), which likely acted as virulence factors. RAP3 was homologous to a beta subunit of propionyl-CoA carboxylase required for the development of Myxococcus xanthus. RAP4 had homology to a quinone oxidoreductase involved in electron transport and ATP formation. RAP5 showed significant homology to a uridine kinase that mediates phosphorylation of uridine and azauridine. RAP6 shared high sequence identity with 3-methyl-2-oxobutanoate-hydroxymethyltransferase (also known as ketopantoate hydroxymethyltransferase or PanB) involved in the operation of the tricarboxylic acid cycle. The findings described here would help to elucidate the molecular mechanisms underlying the differentiation process of B. thuringiensis and unravel novel pathogenic genes.
- MeSH
- Bacillus thuringiensis klasifikace genetika růst a vývoj izolace a purifikace MeSH
- bakteriální proteiny genetika metabolismus MeSH
- DNA fingerprinting metody MeSH
- polymerázová řetězová reakce s reverzní transkripcí metody MeSH
- regulace genové exprese u bakterií MeSH
- spory bakteriální klasifikace genetika růst a vývoj metabolismus MeSH
- vývojová regulace genové exprese MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
This paper concerns the formation of biofilm in bacteria of the genus Arcobacter. A multiplex polymerase chain reaction (PCR) method was introduced and optimized for detecting biofilm while using the intercalating dyes ethidium monoazide (EMA) and propidium monoazide (PMA), first for analysis of strains of the genus Arcobacter from a collection, and then applied to samples of prepared biofilms. The results of the study indicate considerable variability among species of bacteria within the genus Arcobacter. The EMA-PMA PCR method can distinguish viable cells from dead cells and is therefore suitable for determining the viability of cells.
- MeSH
- azidy chemie MeSH
- biofilmy * MeSH
- Campylobacter genetika izolace a purifikace fyziologie MeSH
- interkalátory chemie MeSH
- mikrobiální viabilita * MeSH
- multiplexová polymerázová řetězová reakce metody MeSH
- propidium analogy a deriváty chemie MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
The low frequency of nontuberculous mycobacterial infections, nonspecific symptoms for individual mycobacteria, and the lack of specific identification methods could alter correct diagnosis. This study presents a combined microbiology and molecular-based approach for Mycobacterium marinum detection in four aquarists with cutaneous mycobacterial infection. Simultaneously, ecology screening for M. marinum presence in the aquarists' fish tanks was performed. A total of 38 mycobacterial isolates originated from four human patients (n = 20), aquarium animals (n = 8), and an aquarium environment (n = 10). Isolate identification was carried out using 16S rRNA sequence analysis. A microbiology-based approach, followed by 16S rRNA sequence analysis, was successfully used for detection of M. marinum in all four patients. Animal and environmental samples were simultaneously examined, and a total of seven mycobacterial species were isolated: Mycobacterium chelonae , Mycobacterium fortuitum , Mycobacterium gordonae , Mycobacterium kansasii , Mycobacterium mantenii , Mycobacterium marinum , and Mycobacterium peregrinum . The presence of M. marinum was proven in the aquarium environments of two patients. Although M. marinum is described as being present in water, it was detected only in fish.
- MeSH
- antibakteriální látky farmakologie terapeutické užití MeSH
- atypické mykobakteriální infekce diagnóza farmakoterapie mikrobiologie patologie MeSH
- klarithromycin farmakologie terapeutické užití MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- mikrobiologie životního prostředí MeSH
- Mycobacterium marinum klasifikace účinky léků genetika izolace a purifikace MeSH
- Mycobacterium klasifikace genetika izolace a purifikace MeSH
- RNA ribozomální 16S genetika MeSH
- ryby mikrobiologie MeSH
- výsledek terapie MeSH
- zvířata MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
Vacuoles play an important role in the physiology of pathogenic Candida spp. However, information on Candida albicans vacuolar enzymes, their properties, and regulation is scarce. Expression of the genes APR1 and CPY1 encoding vacuolar aspartic protease and serine carboxypeptidase, respectively, was analyzed using a clinical isolate of C. albicans. The transcription of both APR1 and CPY1 was upregulated in midexponential phase, together with increasing size of the vacuoles, when C. albicans was cultivated in yeast extract-peptone-dextrose agar at 30 °C. However, simultaneous upregulation of protein synthesis occurred only for Cpy1p. Analysis of APR1 and CPY1 expression under nitrogen-limited conditions revealed that the genes were regulated on both the transcriptional and translational levels and detectable amounts of Apr1p were synthesized only when C. albicans was grown in nitrogen-limited media.
- MeSH
- aspartátové proteasy genetika metabolismus MeSH
- Candida albicans enzymologie genetika růst a vývoj MeSH
- dusík metabolismus MeSH
- karboxypeptidasy genetika metabolismus MeSH
- kultivační média metabolismus MeSH
- regulace genové exprese u hub MeSH
- upregulace MeSH
- vakuoly enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of this study was to determine which species of culturable bacteria are associated with ectomycorrhizae (ECM) of Norway spruce (Picea abies (L.) Karst) in the Sudety Mountains, exposed for years to atmospheric pollutants, acid rain, and climatic stress, and to identify particular species that have adapted to those conditions. Biolog identification was performed on bacterial species from ECM of adult spruce trees and seedlings of stands with low, intermediate, and high forest decline. Bacterial diversity in ECM associated with adult spruce trees, seedlings, and seedlings grown on monoliths was calculated; although the expected values appeared to vary widely, no significant differences among sites were observed. Dendrograms based on the identified bacterial species showed that stands with low forest decline clustered separately from the others. Principal component analysis of the normalized data for ECM-associated species showed a clear separation between stands with high forest decline and stands with low forest decline for seedlings and a less evident separation for adult spruce trees. In conclusion, shifts in ECM-associated culturable bacterial populations seem to be associated with forest decline in Norway spruce stands. Some bacterial species were preferentially associated with mycorrhizal roots depending on the degree of forest decline; this was more evident in seedlings where the species Burkholderia cepacia and Pseudomonas fluorescens were associated with, respectively, ECM of the most damaged stands and those with low forest decline.
- MeSH
- Bacteria genetika růst a vývoj izolace a purifikace MeSH
- ekosystém * MeSH
- kořeny rostlin mikrobiologie MeSH
- mykorhiza fyziologie MeSH
- počet mikrobiálních kolonií MeSH
- populační dynamika MeSH
- RNA ribozomální 16S genetika MeSH
- semenáček mikrobiologie MeSH
- smrk mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
High-resolution melting analysis (HRMA) is a fast (post-PCR) high-throughput method to scan for sequence variations in a target gene. The aim of this study was to test the potential of HRMA to distinguish particular bacterial species of the Staphylococcus genus even when using a broad-range PCR within the 16S rRNA gene where sequence differences are minimal. Genomic DNA samples isolated from 12 reference staphylococcal strains (Staphylococcus aureus, Staphylococcus capitis, Staphylococcus caprae, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus intermedius, Staphylococcus saprophyticus, Staphylococcus sciuri, Staphylococcus simulans, Staphylococcus warneri, and Staphylococcus xylosus) were subjected to a real-time PCR amplification of the 16S rRNA gene in the presence of fluorescent dye EvaGreen™, followed by HRMA. Melting profiles were used as molecular fingerprints for bacterial species differentiation. HRMA of S. saprophyticus and S. xylosus resulted in undistinguishable profiles because of their identical sequences in the analyzed 16S rRNA region. The remaining reference strains were fully differentiated either directly or via high-resolution plots obtained by heteroduplex formation between coamplified PCR products of the tested staphylococcal strain and phylogenetically unrelated strain.
- MeSH
- DNA bakterií chemie MeSH
- DNA fingerprinting metody MeSH
- druhová specificita MeSH
- heteroduplexní analýza MeSH
- molekulární sekvence - údaje MeSH
- ribozomální DNA chemie genetika MeSH
- RNA ribozomální 16S chemie genetika MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- sekvenční seřazení MeSH
- senzitivita a specificita MeSH
- Staphylococcus klasifikace genetika MeSH
- tranzitní teplota MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The objective of this study was to determine the incidence of nontuberculous mycobacteria (NTM) in hot water systems of 4 selected hospital settings. The hospitals provided the following types of disinfection for their hot water systems: hydrogen peroxide and silver, thermal disinfection, chlorine dioxide, and no treatment (control). In each building, 6 samples were collected from 5 sites during a 3 month period. NTM were detected in 56 (46.7%) of 120 samples; the CFU counts ranged from 10 to 1625 CFU/L. The detected NTM species were the pathogens Mycobacterium kansasii, Mycobacterium xenopi, and Mycobacterium fortuitum and the saprophyte Mycobacterium gordonae. The most common to be isolated was M. xenopi, which was present in 51 samples. The hot water systems differed significantly in the incidence of NTM. NTM were not detected in the system treated by thermal disinfection, and a relatively low incidence (20% positive samples) was found in the system disinfected with chlorine dioxide. However, a high incidence was found in the control system with no additional disinfection (70% positives) and in the system using hydrogen peroxide and silver (97% positives). Water temperatures above 50 degrees C significantly limited the occurrence of NTM.
