Denaturant gradient gel electrophoresis (DGGE) enables insight into the diversity of the studied microbial communities on the basis of separation of PCR amplification products according to their nucleotide sequence composition. However, the success of the method is accompanied by the inherent appearance of various sequence artifacts that bias the impression of community structure by generating additional bands representing no virtual microbes. PCR-DGGE artifacts require optimization of the method when aiming at the phylogenetic identification of the selected DGGE bands. The aim of our study was to develop a procedure which will increase the reliability of the identification. Samples of rumen fluid were used for the optimization since they contain a complex microbial community that supports the generation of artifactual bands. An optimized procedure following band excision and elution of microbial DNA is proposed including nuclease treatment, selection of DNA polymerase with proofreading activity, and cloning prior to sequencing and identification analysis.

• MeSH
• Rumen microbiology   MeSH
• Bacteria classification   genetics   isolation & purification   MeSH
• Denaturing Gradient Gel Electrophoresis methods   MeSH
• DNA, Bacterial genetics   MeSH
• Phylogeny MeSH
• Bacterial Typing Techniques methods   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• DNA, Bacterial MeSH

The effect of daily supplementation of nisin (2 mg/L), monensin (5.88 mg/L) and nisin and monensin (2 + 5.88 mg/L) on ovine ruminal ciliates and bacteria was investigated using the artificial rumen RUSITEC. Major groups in RUSITEC were Entodinium spp. and Dasytricha ruminantium. The supplementation of nisin significantly increased the population of both major ciliate groups. The supplementation of monensin significantly decreased the population of both groups. The combined effect of nisin and monensin was similar to the effect of monensin. Monensin had strong antiprotozoic effects in contrast to the stimulatory effects of nisin. D. ruminantium followed by Entodinium spp. appeared more resistant to tested compounds than other rumen ciliates. Tested additives did not significantly influence the presence and growth of amylolytic streptococci and enterococci but nisin showed a tendency to decreasing the concentration of Escherichia coli and lactobacilli.

• MeSH
• Rumen microbiology   parasitology   MeSH
• Bacteria drug effects   MeSH
• Disinfectants administration & dosage   MeSH
• Diet methods   MeSH
• Eukaryota drug effects   MeSH
• Monensin administration & dosage   MeSH
• Nisin administration & dosage   MeSH
• Sheep MeSH
• Models, Theoretical MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Disinfectants MeSH
• Monensin MeSH
• Nisin MeSH

Two representative strains of Gram-negative rumen bacteria from the genus Prevotella were used as model organisms in order to evaluate the effect of cinnamaldehyde (the secondary metabolite found in extracts of the Cinnamomum family) vs. sodium monensin on growth, cell size and cell protein production. Prevotella bryantii B(1)4 was found to be remarkably more resistant to the action of both compounds than Prevotella ruminicola 23. The approximate IC(50) concentrations of sodium monensin influenced the increase in cell size of both strains during growth, which was much more pronounced in the case of the B(1)4 strain. A similar effect was observed in strain B(1)4 when 1.438 mmol/L cinnamaldehyde was added to the growth medium, indicating a possible interference with cell division. The action of cinnamaldehyde on P. bryantii B(1)4 was concentration-dependent, in contrast to the effect observed on P. ruminicola 23.

• MeSH
• Acrolein analogs & derivatives   pharmacology   MeSH
• Antiprotozoal Agents pharmacology   MeSH
• Rumen microbiology   MeSH
• Phenotype MeSH
• Inhibitory Concentration 50 MeSH
• Monensin pharmacology   MeSH
• Parasitic Sensitivity Tests MeSH
• Prevotella ruminicola classification   drug effects   growth & development   metabolism   MeSH
• Prevotella classification   drug effects   growth & development   metabolism   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Names of Substances
• Acrolein MeSH
• Antiprotozoal Agents MeSH
• cinnamaldehyde MeSH Browser
• Monensin MeSH

The influence of a host's diet on Butyrivibrio and Pseudobutyrivibrio populations was investigated by competitive PCR. Specific primers were designed and competitive PCRs developed for both groups. Results (from 4 cows with different diets) suggested that high-fiber intake essentially increases the Butyrivibrio amounts in the rumen, whereas high-energy food additives lead to its suppression. The Pseudobutyrivibrio concentration also changed during the experiment but without any significant relation to the host's diet.

• MeSH
• Rumen microbiology   MeSH
• Butyrates metabolism   MeSH
• Butyrivibrio genetics   growth & development   isolation & purification   MeSH
• Clostridium genetics   growth & development   isolation & purification   MeSH
• DNA, Bacterial analysis   MeSH
• Gram-Negative Aerobic Rods and Cocci genetics   growth & development   isolation & purification   MeSH
• Animal Feed MeSH
• Colony Count, Microbial MeSH
• Polymerase Chain Reaction methods   MeSH
• Dietary Fiber MeSH
• RNA, Ribosomal, 16S classification   genetics   MeSH
• Cattle MeSH
• Animals MeSH
• Check Tag
• Cattle MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Butyrates MeSH
• DNA, Bacterial MeSH
• Dietary Fiber MeSH
• RNA, Ribosomal, 16S MeSH

The effect of the oregano essential oil (EO) on lactobacilli and E. coli isolated from chicken was showed in the series of in vitro and in vivo experiments. Minimum inhibitory concentrations of amikacin, apramycin, streptomycin, neomycin against E. coli strains increased after oral application of EO.

• MeSH
• Origanum chemistry   MeSH
• Escherichia coli drug effects   isolation & purification   MeSH
• Feces microbiology   MeSH
• Chickens microbiology   MeSH
• Lactobacillus drug effects   MeSH
• Microbial Sensitivity Tests MeSH
• Plant Oils pharmacology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Plant Oils MeSH

The effect of selenium supplementation on the rumen protozoan population of sheep was demonstrated. Both the total and generic counts of rumen ciliates in sheep fed a diet with basal Se content (70 microg/kg dry matter) were compared to those of animals given feed supplemented with inorganic (disodium selenite) or organic Se (selenized yeast) (310 microg/kg dry matter). The genera of Entodinium, Isotricha, Dasytricha, Ophryoscolex, Diploplastron and Polyplastron occurred in all sheep except for the control, in which Ophryoscolex was not observed. The population of Ophryoscolex caudatus f. tricoronatus was significantly higher in sheep supplemented with organic Se than in animals given inorganic Se (by 160 %). Supplementation of feed with selenized yeast induced significant growth in the Diploplastron population (by 63 %) while no change occurred in sheep given selenite. The populations of Dasytricha ruminantium and Polyplastron multivesiculatum were higher than control in both Se-supplemented groups. The ciliate population of Entodinium spp. was not influenced by Se supplements. Our results suggest a protective effect of Se feed supplementation on the development of some rumen ciliate species in young ruminants.

• MeSH
• Inorganic Chemicals MeSH
• Rumen drug effects   parasitology   MeSH
• Ciliophora drug effects   growth & development   MeSH
• Organic Chemicals administration & dosage   pharmacology   MeSH
• Sheep, Domestic parasitology   MeSH
• Dietary Supplements * MeSH
• Selenium pharmacology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Inorganic Chemicals MeSH
• Organic Chemicals MeSH
• Selenium MeSH