A quantitative TaqMan minor-groove binder real-time PCR assay was developed for the sensitive detection of a ruminant-specific genetic marker in fecal members of the phylum Bacteroidetes. The qualitative and quantitative detection limits determined were 6 and 20 marker copies per PCR, respectively. Tested ruminant feces contained an average of 4.1 x 10(9) marker equivalents per g, allowing the ...

Five samples of soil collected from a lead and zinc mine were used to assess the effect of combined contamination of heavy metals on soil bacterial communities using a polyphasic approach including characterization of isolates by culture method, community level catabolic profiling in BIOLOG GN microplates, and genetic community fingerprinting by denaturing gradient gel electrophoresis of 16S rD...

The changes of ileal bacterial microbiota in pigs during the first 2 weeks post-weaning in response to feeding lincomycin, organic acids or herbal extract were characterized using 16S rRNA gene-based PCR-denaturing gradient gel electrophoresis (DGGE) profiling, DNA sequencing, and real-time PCR (QPCR) techniques. Both time post-weaning and the dietary treatments resulted in a shift of the micro...

Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting was recently introduced into food microbiology. This paper describes the technique and reports on the state-of-the-art application of this technique to food and food-related ecosystems. Applications of PCR-DGGE in several fields of food microbiology are reviewed: the identification of microorganisms isol...

METHODS. Monodispersed cell suspensions obtained by enzymatic digestion of conjunctival tissue were applied to a single-density Percoll gradient. Epithelial cells obtained from the top layer of the gradient were cultured to confluence. Mast cells obtained from the pellet were equilibrated in culture medium and further purified using a two-step Percoll gradient. Using reverse transcription polym...

The intestinal bacteria community structure and diversity of the Oriental armyworm, Mythimna separata (Walker) (Lepidoptera: Noctuidae), was studied by analysis of a 16S rDNA clone library, denaturing gradient gel electrophoresis,and culture-dependent techniques. The 16S rDNA clone library revealed a bacterial community diversity comprising Cyanobacteria, Firmicutes, Actinobacteria, Gracilicute...

Methods that enabled the identification, detection, and enumeration of Bifidobacterium species by PCR targeting the transaldolase gene were tested. Bifidobacterial species isolated from the feces of human adults and babies were identified by PCR amplification of a 301-bp transaldolase gene sequence and comparison of the relative migrations of the DNA fragments in denaturing gradient gel electro...

Candida albicans is an opportunistic pathogen. Its proliferation in human hosts is believed to be controlled by immunologic mechanisms. The plasma membrane of the fungus posseses an H+-ATPase (PM-ATPase) which actively extrudes protons to generate an electrochemical gradient which is used in co-transport of nutrients. This ATPase is associated with the growth, dimorphism and pathogenicity of th...

Soil microorganisms play an important role in agricultural ecosystem. However, there is little information about the effects of putative allelochemicals on specific soil vivo. Cucumber seedlings were treated with four concentrations ferulic and p-hydroxybenzoic acids (0-1.0 μmol g-1 soil) soil. Effects rhizosphere Fusarium community structures abundance analyzed by polymerase chain reaction den...