Microbiota of sulfur-rich environments has been extensively studied due to the biotechnological potential of sulfur bacteria, or as a model of ancient life. Cold terrestrial sulfur springs are less studied compared to sulfur-oxidizing microbiota of hydrothermal vents, volcanic environments, or soda lakes. Despite that, several studies suggested that sulfur springs harbor diverse microbial communities because of the unique geochemical conditions of upwelling waters. In this study, the microbiota of five terrestrial sulfur springs was examined using a 16 S rRNA gene sequencing. The clear dominance of the Proteobacteria and Campylobacterota phyla of cold sulfur springs microbiota was observed. Contrary to that, the microbiota of the hot sulfur spring was dominated by the Aquificota and Firmicutes phylum respectively. Sulfur-oxidizing genera constituted a dominant part of the microbial populations with the Thiothrix and Sulfurovum genera identified as the core microbiota of cold sulfur terrestrial springs in Slovakia. Additionally, the study emphasizes that sulfur springs in Slovakia support unique, poorly characterized bacterial communities of sulfur-oxidizing bacteria.

• Klíčová slova
• Microbiota, Slovakia, Sulfur springs, Sulfur-oxidizing bacteria,
• Publikační typ
• časopisecké články MeSH

During characterization of autochthonic Vyhna travertine source microflora, several bacterial strains were isolated and characterised. Isolate T6, a halotolerant, moderately alkaliphilic and thermophilic bacterial isolate, was further characterised based on physiological, microbiological and biochemical tests and phylogenetic 16S rRNA analysis. On the basis of the results obtained, the T6 isolate should be placed in the genus Oceanobacillus, and it is probably a prototype of a novel bacterial species. Characterization of the T6 isolate broadens our knowledge on variability of halophilic bacteria of Oceanobacillus genus and expands data on travertine-associated bacterial communities.

• MeSH
• Bacillaceae klasifikace   genetika   izolace a purifikace   fyziologie   MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• mikroskopie MeSH
• molekulární sekvence - údaje MeSH
• přírodní prameny mikrobiologie   MeSH
• ribozomální DNA chemie   genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• shluková analýza MeSH
• techniky typizace bakterií MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA bakterií MeSH
• ribozomální DNA MeSH
• RNA ribozomální 16S MeSH

The chitinase gene was molecularly characterized in five Bacillus thuringiensis Mexican isolates, MR10, MR11, MR21, MR33, and RN52. The proteins derived from these genes were tested for their chitinase activity using fluorogenic chitin derivatives. In order to verify if chitinase genes were functional, they were cloned, and enzymatic activity of recombinant chitinases was also tested. Results indicated that enzymes exhibited endochitinase activity. The highest hydrolytic activity shown against the chitin tetrameric derivative occurred at pH value of 6.5, and the optimum activity temperature was around 60 °C. The recombinant endochitinases showed a molecular mass of ∼77 kDa with isoelectric points from 6.5 to 7.0. Analysis of the nucleotide sequences showed highly conserved sequences among all isolates (97-99 %). Gene sequence analysis revealed a putative promoter (-35 TTGAGA and -10 TTAATA) and a Shine-Dalgarno sequence (5´-AGGAGA-3´) upstream from the open reading frame. The deduced amino acid sequence revealed that the proteins are modular enzymes composed by a family 18 glycosyl hydrolase domain located between amino acids 134 and 549, a fibronectin-binding domain (580 through 656), and a chitin-binding domain (664 through 771). The deduced amino acid sequences of our isolates showed a similarity close to 100 % respect to the sequences reported in the GenBank database.

• MeSH
• Bacillus thuringiensis enzymologie   genetika   izolace a purifikace   MeSH
• chitin metabolismus   MeSH
• chitinasy chemie   genetika   metabolismus   MeSH
• DNA bakterií chemie   genetika   MeSH
• exprese genu MeSH
• izoelektrický bod MeSH
• klonování DNA MeSH
• koncentrace vodíkových iontů MeSH
• molekulární sekvence - údaje MeSH
• molekulová hmotnost MeSH
• otevřené čtecí rámce MeSH
• promotorové oblasti (genetika) MeSH
• sekvence aminokyselin MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie aminokyselin MeSH
• sekvenční seřazení MeSH
• stabilita enzymů MeSH
• teplota MeSH
• vazebná místa MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Mexiko MeSH
• Názvy látek
• chitin MeSH
• chitinasy MeSH
• DNA bakterií MeSH

Actinobacteria (Actinomycetes) are a significant and interesting group of gram-positive bacteria. They are regular, though infrequent, members of the microbial life in the rumen and represent up to 3 % of total rumen bacteria; there is considerable lack of information about ecology and biology of rumen actinobacteria. During the characterization of variability of rumen treponemas using non-cultivation approach, we also noted the variability of rumen actinobacteria. By using Treponema-specific primers a specific 16S rRNA gene library was prepared from cow and sheep rumen total DNA. About 10 % of recombinant clones contained actinobacteria-like sequences. Phylogenetic analyses of 11 clones obtained showed the high variability of actinobacteria in the ruminant digestive system. While some sequences are nearly identical to known sequences of actinobacteria, we detected completely new clusters of actinobacteria-like sequences, representing probably new, as yet undiscovered, group of rumen Actinobacteria. Further research will be necessary for understanding their nature and functions in the rumen.

• MeSH
• Actinobacteria klasifikace   genetika   izolace a purifikace   MeSH
• bachor mikrobiologie   MeSH
• biodiverzita MeSH
• DNA bakterií genetika   MeSH
• fylogeneze MeSH
• molekulární sekvence - údaje MeSH
• ovce MeSH
• ribozomální DNA genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA bakterií MeSH
• ribozomální DNA MeSH
• RNA ribozomální 16S MeSH

Small plasmid pKST23 was isolated from sheep ruminal Escherichia coli population. Plasmid sequence was determined to be 2,779 bp in length and was found to have an overall 42 % of GC pairs. However, its sequence can be divided into two regions based on genetic composition and the GC content. It was found that the high GC region spanning approximately from nucleotide 1,300 to 2,750 was identical to a group of small Escherichia coli plasmids and encoded a putative replication protein identical to plasmid pKL1 Rep protein. The part with lower GC pairs seemed to be more specific as it showed no similarity to the GenBank database. Computational analysis revealed four open reading frames, two of which showed considerable homology to replication proteins. PCR primers targeting parts of the two different regions of plasmid pKST23 were used to assess the occurrence of related plasmids within ruminal E. coli population.

• MeSH
• bachor mikrobiologie   MeSH
• Escherichia coli chemie   genetika   izolace a purifikace   MeSH
• molekulární sekvence - údaje MeSH
• otevřené čtecí rámce MeSH
• ovce MeSH
• plazmidy chemie   genetika   MeSH
• proteiny z Escherichia coli chemie   genetika   MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční homologie aminokyselin MeSH
• zastoupení bazí MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• proteiny z Escherichia coli MeSH

Complete 16S rRNA sequences were determined of recently proposed new species of treponemes designated strain S and T. Sequence comparison indicated that both species belong to the Treponema saccharophilum cluster, having thus at least 5 cultivable representatives. Phylogenetic analysis of available GenBank 16S rRNA sequences revealed two phylogenetically distant treponema clusters (T. saccharophilum cluster and T. bryantii cluster). Surprisingly, while among cultivated treponemes dominate T. saccharophilum cluster members, detailed analysis showed that all treponema-like sequences obtained by culture independent 16S rRNA methods belong to the T. bryantii cluster, from which only two cultivable representatives have so far been known. Meta-analysis of available data revealed that treponemes are an infrequent and minor group of bacteria, representing less than 2.4% of total rumen bacteria.

• MeSH
• bachor mikrobiologie   MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• molekulární sekvence - údaje MeSH
• počet mikrobiálních kolonií MeSH
• přežvýkavci mikrobiologie   MeSH
• ribozomální DNA chemie   genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• shluková analýza MeSH
• Treponema klasifikace   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA bakterií MeSH
• ribozomální DNA MeSH
• RNA ribozomální 16S MeSH

The inter- and intraspecies variability of lactate dehydrogenase (ldh) gene was determined among the predominant ruminal lactate utilizing bacteria. Nearly complete nucleotide sequences of ldh gene, encoding NAD-dependent lactate dehydrogenase of three Megasphaera elsdenii and six Selenomonas ruminantium strains, were obtained and compared. Phylogenetic analyses revealed a limited variability between the ldh sequences studied. The majority of differences observed were silent mutations at the 3rd position of codons. Surprisingly, the intraspecies diversity of the ldh gene among S. ruminantium isolates was higher than the interspecies level between S. ruminantium and M. elsdenii, which strongly suggests the possibility of acquisition of this gene by horizontal gene transfer.

• MeSH
• bachor mikrobiologie   MeSH
• bakteriální proteiny genetika   MeSH
• bodová mutace MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• kyselina mléčná metabolismus   MeSH
• L-laktátdehydrogenasa genetika   MeSH
• Megasphaera enzymologie   genetika   izolace a purifikace   metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie MeSH
• Selenomonas enzymologie   genetika   izolace a purifikace   metabolismus   MeSH
• shluková analýza MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bakteriální proteiny MeSH
• DNA bakterií MeSH
• kyselina mléčná MeSH
• L-laktátdehydrogenasa MeSH

The diversity of archaebacteria associated with anaerobic rumen protozoan Entodinium caudatum in long term in vitro culture was investigated by denaturing gradient gel electrophoresis (DGGE) analysis of hypervariable V3 region of archaebacterial 16S rRNA gene. PCR was accomplished directly from DNA extracted from a single protozoal cell and from total community genomic DNA and the obtained fingerprints were compared. The analysis indicated the presence of a solitary intensive band present in Entodinium caudatum single cell DNA, which had no counterparts in the profile from total DNA. The identity of archaebacterium represented by this band was determined by sequence analysis which showed that the sequence fell to the cluster of ciliate symbiotic methanogens identified recently by 16S gene library approach.

• MeSH
• anaerobióza MeSH
• Archaea klasifikace   genetika   růst a vývoj   izolace a purifikace   MeSH
• bachor mikrobiologie   parazitologie   MeSH
• Ciliophora genetika   růst a vývoj   izolace a purifikace   MeSH
• DNA archebakterií analýza   izolace a purifikace   MeSH
• druhová specificita MeSH
• fylogeneze MeSH
• kultivační média MeSH
• molekulární sekvence - údaje MeSH
• ovce MeSH
• protozoální DNA analýza   izolace a purifikace   MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• symbióza MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA archebakterií MeSH
• kultivační média MeSH
• protozoální DNA MeSH
• RNA ribozomální 16S MeSH

Genome analysis of Treponema zioleckii proved that, in this bacterium, besides chromosomal DNA, a relatively small extrachromosomal DNA element is present. This element was shown to be a double-stranded circular plasmid DNA of approximately 7 kbp; it was designated as pKT. The plasmid was characterized by molecular and bioinformatic analysis. No pKT homologous DNA sequences were detected in other rumen Treponema strains. The overall G+C content of the pKT plasmid is approximately 56 %, which is higher than in other Treponema plasmids or genomes. The Rep module of the pKT plasmid consisting of the rep gene and the region of repeats was identified within a 1.6-kbp fragment. The putative rep gene encodes the replication protein belonging to the pfam04796 RepA_C family of proteins with the highest similarity (25 % within 249 amino acids) to the RepA protein from the green sulfur bacterium Prosthecochloris aestuarii.

• MeSH
• bachor mikrobiologie   MeSH
• bakteriální proteiny genetika   MeSH
• DNA bakterií analýza   izolace a purifikace   MeSH
• fylogeneze MeSH
• genom bakteriální * MeSH
• hybridizace nukleových kyselin MeSH
• molekulární sekvence - údaje MeSH
• ovce MeSH
• plazmidy genetika   MeSH
• replikace DNA MeSH
• sekvence aminokyselin MeSH
• sekvenční analýza DNA MeSH
• Treponema genetika   izolace a purifikace   MeSH
• zastoupení bazí MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bakteriální proteiny MeSH
• DNA bakterií MeSH

Large Enterococcus faecalis F4 bacteriophage (described earlier) consisting of double-stranded linear DNA of approximately 60 kb was characterized. Library was prepared of its random DNA fragments and selected recombinants were sequenced. Three phage essential genes were characterized: DNA polymerase, replicative DNA helicase and a minor capsid protein, showing only limited homology to other known phage encoded genes. The occurrence of these genes among enterococci was determined by PCR method. Only two out of 40 tested isolates possessed all three genes, another three isolates contained at least one of the genes, demonstrating low frequency F4 lysogens among natural enterococcal isolates.

• MeSH
• bakteriofágy klasifikace   genetika   fyziologie   MeSH
• Enterococcus faecalis virologie   MeSH
• genom virový MeSH
• lyzogenie MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce MeSH
• sekvenční analýza DNA MeSH
• virové proteiny genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• virové proteiny MeSH