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P Javorský

Publications and source records attributed to P Javorský.

At least 19 recordsLinked to original sources

Specific heat of delta-Pu stabilized by Am.

Detailed specific heat C(p) measurements of delta-Pu stabilized by Am (8%-20%) were performed in the temperature range 4.5-300 K. The coefficient of the electronic specific heat gamma, which reflects the quasiparticle density of states at the Fermi level E(F), is smaller than originally assumed and, depending on the estimate of phonon contributions, a value between 35 and 55 mJ/mol K2 can be deduced for Pu-8% Am. For higher Am concentrations, which expand the lattice, gamma decreases slightly with the Am content. An applied magnetic field of 9 T had no effect on C(p). The results strongly suggest that itinerant 5f states at E(F) are not appropriate for describing delta-Pu.

Journal Article↗

Spreading and mutability of Selenomonas ruminantium plasmids.

Two small plasmids from Selenomonas ruminantium strain 19D were cloned in Escherichia coli and completely characterized. Sequence comparison indicated that the plasmids are similar to those reported in genetically vaguely related S. ruminantium strain S20. Small 1.4-kb plasmids pSRD191 and pONE430 are only distantly related (approximately 30 % for deduced Rep protein amino acid sequence) but possess a short highly conserved region outside rep gene. Larger plasmids pSRD192 and pONE429 possess large identical DNA regions in an otherwise dissimilar background. Recombination is proposed as an important mechanism of evolution and spreading of S. ruminantium plasmids.

Animals↗

Limited genetic variability in Megasphaera elsdenii strains.

Levels of phenotypic and genotypic diversity among seven Megasphaera elsdenii strains recovered from rumen contents of cattle, sheep and lambs were determined by a combination of antibiotic-resistance analysis and PCR fingerprint techniques targeted both to the ribosomal RNA operon (ARDRA, RISA) and the whole genome (ERIC-PCR, RAPD-PCR). Despite exhibiting different antibiotic resistance profiles, the tested strains represent genetically nearly identical isolates. Close genetic relatedness was found among M. elsdenii isolates that originated from vastly different habitats worldwide, as revealed by the comparison of 16S rDNA sequences.

Animals↗

New species of rumen treponemes.

Three strains of rumen treponemes were isolated and partially characterized. The strains differed significantly one from another in morphology, fermentation characteristics and plasmid profiles. Their genetic variability was assayed using DNA-based molecular approaches. Easily differentiated ARDRA (amplified ribosomal DNA restriction analysis) patterns indicated that the strains represent different bacterial species.

Animals↗

Mercuric reductase gene transfer from soil to rumen bacteria.

Conjugal transfer between soil bacterial population and microorganisms isolated from the rumen of herbivores from mercury-polluted area was investigated. The transfer of merA encoding mercury-resistance plasmids from soil bacteria Enterobacter cloacae and Enterococcus durans into two ruminal isolates Citrobacter freundii and Bacillus subtilis was observed. Approximately the same frequency of mobilization in mating experiments was observed for both Gram-negative (approximately 2.5 x 10(-8), transconjugants-to-recipient ratio) and Gram-positive (approximately 1.3 x 10(-8)) bacteria.

Animals↗

Seasonal dynamics of antibiotic-resistant Enterobacteriaceae in the gastrointestinal tract of domestic sheep.

Considerable variation in counts of antibiotic-resistant enterobacteria in the ovine gastrointestinal tract was observed. The occurrence of ruminal and fecal isolates resistant to ampicillin (Ap), kanamycin (Km) and tetracycline (Tc) culminated in summer months, followed by rapid decline in subsequent months. Using PCR the tem1bla (Apr), aphA1 (Kmr) and tetB (Tcr) genes were found to be predominant. Under in vitro conditions all resistance genes were transferable into laboratory Escherichia coli strain with relatively high frequency (10(-3) transconjugants per recipient).

Animals↗

Bacteriocin production and sensitivity.

Using an overlay test the production of bacteriocin-like activity and resistance was found in 6 of the total of 7 isolates (5 enterococcal and 2 streptococcal). The majority of strains were sensitive to all bacteriocin producers tested. After acetone precipitation, bacteriocin precipitates were tested for thermal stability. They exhibited high stability at 37 degrees C and some of them were active even after a treatment at 95 degrees C.

Anti-Bacterial Agents↗

Occurrence of plasmid-mediated ampicillin resistance among enterobacteria from the ovine rumen.

Seasonal samplings of rumen and fecal populations of Enterobacteriacae from sheep digestive tract were done to elucidate potential occurrence and spreading of antibiotic resistance in the environment. Thus 350 rumen and fecal isolates were tested for ampicillin (Amp) resistance in single sampling. Low frequency of Amp resistance (from 0 to 15%) was observed. The occurrence of tem1 encoded Amp resistance confirmed by PCR was observed among both rumen and fecal isolates. The small tem1 carrying plasmid and its transfer (mobilization) was detected and partially characterized after conjugation to laboratory Escherichia coli strain.

Ampicillin↗

GATC-specific restriction--modification systems in ruminal bacteria.

The GATC-specific restriction and modification activities were analyzed in 11 major bacterial representatives of ruminal microflora. Modification phenotype was observed in 13 out of 40 ruminal strains. MboI isoschizomeric restriction endonucleases were detected in 10 bacterial strains tested; three strains lacked any detectable corresponding endonuclease activity. The only examined strain of Mitsuokella multi-acida was found to possess a different type of endonuclease activity. This is the first report on restriction activity in ruminal treponemes M. multiacida and Megasphaera elsdenii.

Animals↗

Are ruminal bacteria protected against environmental stress by plant antioxidants?

AIMS: To investigate the activity response of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) of the rumen bacterium Streptococcus bovis following exposure to mercury(II) chloride (HgCl(2) in the presence of plant antioxidants. METHODS AND RESULTS: Streptococcus bovis was grown with 0 or 5 microg ml(-1) of HgCl(2) alone or together with antioxidant substances (AOS): seleno-l-methionine (Se), alpha-tocopherol (alpha toc), beta-carotene (beta car), melatonin (mel). The activities of SOD and GHPx were estimated in supernatants of disrupted bacterial cells. A significant decrease in the Strep. bovis SOD activity in the presence of HgCl(2) and tested AOS, except mel, was observed. The GSHPx activity of Strep. bovis was under the same cultivation conditions nonsignificantly changed and a significant decrease in the GSHPx activity was recorded only in the presence of beta car. CONCLUSIONS: The positive effect of Se, alpha toc and beta car on the elimination of environmental stress, evoked by mercury, in ruminal bacterium Strep. bovis in vitro was documented. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential role of plant antioxidants in elimination of the environmental stress of ruminal bacteria evoked by heavy metals is discussed.

Animals↗

Plasmids of Selenomonas ruminantium and development of host-vector system.

A high frequency of plasmids was detected in the rumen bacterium Selenomonas ruminantium. Plasmids 0.9-20 kb in size were detected in more than 50% tested strains. Densitometric analysis indicated that plasmid DNA could represents more than 25% of total cellular DNA. Up to six plasmids were detected in strain S. ruminantium 18. Two smallest cryptic plasmids pSRD181 and pSRD182 from this strain were cloned into Escherichia coli vector pBluescriptSK+ and partially characterized. The plasmid pSRD181 is 1.4 kb and pSRD182 is 2.0 kb. While computer analysis of pSRD181 sequence data showed high homology with replication protein of Staphylococcus aureus plasmids, the pSRD182 sequence showed no significant homology in GenBank data. Strain S. ruminantium 28 was successfully transformed with pJW1 derived plasmid pJ1B1 using ampicillin resistance gene as marker. This is the first report on transformation of selenomonads with foreign DNA.

Ampicillin Resistance↗

Bacteriocins of ruminal bacteria.

Similar sequences of distribution of structural genes encoding enterocin A (isolated from the ruminal strain E. faecium BC25) and enterolysin A (isolated from the ruminal amylolytic strain S. bovis II/1) were demonstrated by PCR using oligonucleotide primers specific for these bacteriocins within the ruminal enterococcal and streptococcal strains. Variable occurrence of these bacteriocins was found within the populations of Gram-positive ruminal cocci.

Animals↗

Restriction and modification systems of ruminal bacteria.

A high frequency of type II restriction endonuclease activities was detected in Selenomonas ruminantium but not in other rumen bacteria tested. Eight different restriction endonucleases were characterized in 17 strains coming from genetically homogeneous local population. Chromosomal DNA isolated from S. ruminantium strains was found to be refractory to cleavage by various restriction enzymes, implying the presence of methylase activities additional to those required for protection against the cellular endonucleases. The presence of Dam methylation was detected in S. ruminantium strains as well as in several other species belonging to the Sporomusa subbranch of low G + C Gram-positive bacteria (Megasphaera elsdenii, Mitsuokella multiacidus).

Animals↗

Isolation and characterization of enterocin BC25 and occurrence of the entA gene among ruminal gram-positive cocci.

Enterocin BC25, a bacteriocin produced by Enterococcus faecium BC25 isolated from the rumen of cow was purified to homogeneity and sequenced. Twenty amino acids were identified in the peptide chain (TTHSGKYYGNGVYCT-KNKCT), identical to the N-terminal sequence of enterocin A. The DNA sequence of the enterocin BC25 structural gene and putative immunity protein exhibited high similarity to the entA gene. The occurrence of a 726 bp amplicon containing the enterocin A structural gene was studied among gram-positive ruminal cocci by PCR. Our results showed wide occurrence of the entA structural gene among ruminal enterococcal and streptococcal bacterial strains tested, and indicate variable ability to express bacteriocin production and resistance.

Amino Acid Sequence↗

Lack of GATC sites in the genome of Streptococcus bovis bacteriophage F4.

A strong bias against GATC sites was observed in the genome of phage F4, a lytic Streptococcus bovis bacteriophage. Only three GATC sites were found within the 60.4-kbp genome of this phage. The comparative lack of GATC sequences within the F4 genome was probably not due to dam methylation, as no modification within this site was detected using methylation-sensitive isoschizomer pair restriction endonuclease analysis. The short oligonucleotide composition of available S. bovis DNA sequences suggested the existence of an unknown mechanism for counterselection of GATC sites in S. bovis bacteriophages.

Animals↗

Partial purification and characterization of RalF40I, a class II restriction endonuclease from Ruminococcus albus F-40, which recognizes and cleaves 5'-/GATC-3'.

Restriction endonuclease RalF40I was purified from cell-free extracts of the rumen cellulolytic bacterium Ruminococcus albus F-40 heparin-Sepharose chromatography. The preparation was active only on DNA substrates that were not Dammethylated. RalF401 recognizes the 4-bp palindrome, 5'-/GATC-3', and cleaves DNA at the 5' side of G in the sequence, producing 5' tetranucleotide protruding ends. RalF40I is a class II restriction endonuclease and an isoschizomer of MboI and DpnII.

Animals↗

Lack of surface receptors not restriction-modification system determines F4 phage resistance in Streptococcus bovis II/1.

The resistance of Streptococcus bovis strain II/1, the producer of SbvI restriction endonuclease, to F4 phage infection was demonstrated by the double-agar-layer method. Despite the presence of restriction endonuclease SbvI which can cleave F4 phage DNA to numerous fragments in vitro, the evidence that adsorption inhibition is the most important defence mechanism in phage resistance of S. bovis II/1 strain was obtained by adhesion experiments in vivo. Electron microscopy of phage-host mixtures showed many phage particles on the bacterial surface of phage-sensitive S. bovis 47/3 control strain in comparison with no phage particles seen on S. bovis II/1 (phage-resistant) strain surface.

DNA Restriction-Modification Enzymes↗

A bacteriocin-mediated antagonism by Enterococcus faecium BC25 against ruminal Streptococcus bovis.

A bacteriocin-like activity produced by Enterococcus faecium BC25, isolated from the the rumen of a cow, was partially purified and characterized. The active substance was prepared by ammonium sulfate and chloroform/methanol precipitation of culture supernatant. The bacteriocin was a protein of molecular mass 17.5 kDa. Activity was inactivated by trypsin and proteinase K. The bacteriocin BC25 inhibited growth of amylolytic ruminal strains of Streptococcus bovis, including S. bovis AO 24/85. Results showed that bacteriocine substance BC25 has a bacteriostatic effect when present in concentrations exceeding 125 AU/ml. Agar overlays and batch culture growth experiments proved that E. faecium BC25 was producing bacteriocin that inhibited the growth of S. bovis.

Ammonium Sulfate↗