Gram-stain-positive, catalase and oxidase-negative and short rod-shaped bacterium C10 with occasional branching was isolated under strictly anaerobic conditions from the rumen fluid of a red deer (Cervus elaphus) in the course of study attempting to uncover new xylanolytic and cellulolytic rumen bacteria inhabiting the digestive tract of wild ruminants in the Czech Republic. The anaerobic M10 medium containing bovine rumen fluid and carboxymethylcellulose as a defined source of organic carbon was used in the process of bacterial isolation. The 16S rRNA gene similarity revealed recently characterized new species Actinomyces succiniciruminis Am4T (GenBank accession number of the gene retrieved from the complete genome: LK995506) and Actinomyces glycerinitolerans G10T (GenBank accession number from the complete genome: NZFQTT01000017) as the closest relatives (99.7 and 99.6% gene pairwise identity, respectively), followed by the Actinomyces ruminicola DSM 27982T (97.2%, in all compared fragment of 41468 pb). Due to the taxonomic affinity of the examined strain to both species A. succiniciruminis and A. glycerinitolerans, its taxonomic status towards these species was evaluated using variable regions of rpsA (length of 519 bp) and rplB (597 bp) gene sequences amplified based on specific primers designed so as to be applicable in differentiation, classification, and phylogeny of Actinomyces species/strains. Comparative analyses using rpsA and rplB showed 98.5 and 97.9% similarities of C10 to A. succiniciruminis, respectively, and 97.5 and 97.6% similarities to A. glycerinitolerans, respectively. Thus, gene identities revealed that the evaluated isolate C10 (=DSM 100236 = LMG 28777) is a little more related to the species A. succiniciruminis isolated from the rumen of a Holstein-Friesian cow than A. glycerinitolerans. Phylogenetic analyses confirmed affinity of strain C10 to both recently characterized species. Unfortunately, they did not allow the bacterial strain to be classified into a particular species. Phenotypic characterization suggested similar conclusions. This brief contribution is aimed at classification and detailed phenotypic characterization of bacterial strain C10 isolated from the rumen of a wild red deer exhibiting, from the point of view of Actinomyces species, noteworthy cellulolytic and xylanolytic activities.

• MeSH
• Actinomyces klasifikace   genetika   izolace a purifikace   metabolismus   MeSH
• bachor mikrobiologie   MeSH
• bakteriální geny genetika   MeSH
• celulosa metabolismus   MeSH
• DNA bakterií genetika   MeSH
• fenotyp MeSH
• fylogeneze MeSH
• mastné kyseliny analýza   MeSH
• peptidoglykan analýza   MeSH
• RNA ribozomální 16S genetika   MeSH
• vysoká zvěř mikrobiologie   MeSH
• xylany metabolismus   MeSH
• zastoupení bazí MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• celulosa MeSH
• DNA bakterií MeSH
• mastné kyseliny MeSH
• peptidoglykan MeSH
• RNA ribozomální 16S MeSH
• xylany MeSH

The Bifidobacteria show great diversity in the cell surface architecture which may influence the physicochemical properties of the bacterial cell and strain specific properties. The immunomodulatory role of bifidobacteria has been extensively studied, however studies on the immunoreactivity of their protein molecules are very limited. Here, we compared six different methods of protein isolation and purification and we report identification of immunogenic and immunoreactive protein of two human Bifidobacterium longum ssp. longum strains. We evaluated potential immunoreactive properties of proteins employing polyclonal sera obtained from germ free mouse, rabbit and human. The protein yield was isolation method-dependent and the reactivity of proteins detected by SDS-PAGE and Western blotting was heterogeneous and varied between different serum samples. The proteins with the highest immunoreactivity were isolated, purified and have them sequenced. Among the immunoreactive proteins we identified enolase, aspartokinase, pyruvate kinase, DnaK (B. longum ssp. longum CCM 7952) and sugar ABC transporter ATP-binding protein, phosphoglycerate kinase, peptidoglycan synthethase penicillin-binding protein 3, transaldolase, ribosomal proteins and glyceraldehyde 3-phosphate dehydrogenase (B. longum ssp. longum CCDM 372).

• Klíčová slova
• Bifidobacterium, immunoreactivity, moonlighting proteins, probiotics, surface proteins,
• Publikační typ
• časopisecké články MeSH

Social honey bees, Apis mellifera, host a set of distinct microbiota, which is similar across the continents and various honey bee species. Some of these bacteria, such as lactobacilli, have been linked to immunity and defence against pathogens. Pathogen defence is crucial, particularly in larval stages, as many pathogens affect the brood. However, information on larval microbiota is conflicting. Seven developmental stages and drones were sampled from 3 colonies at each of the 4 geographic locations of A. mellifera carnica, and the samples were maintained separately for analysis. We analysed the variation and abundance of important bacterial groups and taxa in the collected bees. Major bacterial groups were evaluated over the entire life of honey bee individuals, where digestive tracts of same aged bees were sampled in the course of time. The results showed that the microbial tract of 6-day-old 5th instar larvae were nearly equally rich in total microbial counts per total digestive tract weight as foraging bees, showing a high percentage of various lactobacilli (Firmicutes) and Gilliamella apicola (Gammaproteobacteria 1). However, during pupation, microbial counts were significantly reduced but recovered quickly by 6 days post-emergence. Between emergence and day 6, imago reached the highest counts of Firmicutes and Gammaproteobacteria, which then gradually declined with bee age. Redundancy analysis conducted using denaturing gradient gel electrophoresis identified bacterial species that were characteristic of each developmental stage. The results suggest that 3-day 4th instar larvae contain low microbial counts that increase 2-fold by day 6 and then decrease during pupation. Microbial succession of the imago begins soon after emergence. We found that bacterial counts do not show only yearly cycles within a colony, but vary on the individual level. Sampling and pooling adult bees or 6th day larvae may lead to high errors and variability, as both of these stages may be undergoing dynamic succession.

• MeSH
• Bacteria klasifikace   genetika   izolace a purifikace   MeSH
• denaturační gradientová gelová elektroforéza MeSH
• DNA bakterií genetika   MeSH
• ekosystém MeSH
• gastrointestinální trakt mikrobiologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• Lactobacillaceae genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• střevní mikroflóra * MeSH
• včely embryologie   růst a vývoj   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA bakterií MeSH
• RNA ribozomální 16S MeSH

The occurrence and species distribution of bifidobacteria in the digestive tract of important representatives of social insects such as ants, bees, wasps and bumblebees as well as the incidence of bifidobacteria in fecal samples of several species of vertebrates represented mainly by reptiles was assigned by culture-independent method based on DGGE and real time PCR. Bifidobacteria were present in the gut of most social insects--honey bees, wasps, cockroaches and bumblebees, except for ants. In honey bees, where the counts of bifidobacteria ranged from 2 to 8% of the total bacteria, the most common species seemed to be Bifidobacterium indicum. Proportion of bifidobacteria was found in broad range from 0.1 to 35-37% in wasps and cockroaches; the variance of bifidobacteria in bumblebees was lower, ranging from 1 to 7% of total bacterial count. Among studied vertebrates, the detectable presence of bifidobacteria was found only in trout (1.1%) and geckos (0.2%), but large amount of these bacteria was observed in Vietnamese box turtle, where bifidobacteria represented nearly one-fourth (22%) of total bacterial counts.

• MeSH
• Bifidobacterium genetika   izolace a purifikace   MeSH
• denaturace nukleových kyselin MeSH
• DNA bakterií genetika   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• feces mikrobiologie   MeSH
• hmyz mikrobiologie   MeSH
• plazi mikrobiologie   MeSH
• polymerázová řetězová reakce MeSH
• ribozomální DNA genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• ryby mikrobiologie   MeSH
• sekvenční analýza DNA 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