We isolated and identified functional groups of bacteria in the rumen of Creole goats involved in ruminal fermentation of native forage shrubs. The functional bacterial groups were evaluated by comparing the total viable, total anaerobic, cellulolytic, hemicellulolytic, and amylolytic bacterial counts in the samples taken from fistulated goats fed native forage diet (Atriplex lampa and Prosopis flexuosa). Alfalfa hay and corn were used as control diet. The roll tubes method increased the possibility of isolating and 16S rDNA gene sequencing allowed definitive identification of bacterial species involved in the ruminal fermentation. The starch and fiber contents of the diets influenced the number of total anaerobic bacteria and fibrolytic and amylolytic functional groups. Pseudobutyrivibrio ruminis and Pseudobutyrivibrio xylanivorans were the main species isolated and identified. The identification of bacterial strains involved in the rumen fermentation helps to explain the ability of these animals to digest fiber plant cell wall contained in native forage species.

• MeSH
• bachor mikrobiologie   MeSH
• bakteriální nálož MeSH
• dieta metody   MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• grampozitivní bakterie klasifikace   genetika   izolace a purifikace   metabolismus   MeSH
• kozy MeSH
• krmivo pro zvířata * MeSH
• molekulární sekvence - údaje MeSH
• ribozomální DNA chemie   genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• shluková analýza 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

Pseudobutyrivibrio xylanivorans strain Mz5(T), an anaerobic bacterium (originating from the rumen of a Holstein-Friesian cow), has some attributes that make it a possible probiotic strain (very active hydrolases, bacteriocin and conjugated linoleic acid production). For the estimation of its adhesion ability, the adhesion test on Caco-2 cells was introduced and adapted. The adhesion was performed in an anaerobic glove box in standard 24-well plates at neutral pH for 30 min. The best method for separation of the adhered bacteria from Caco-2 cells appeared to be homogenization with an automatic pipette. The number of adhered bacteria was too small to be determined microscopically, so a new approach, i.e. detection of the apparent lag phase in liquid growth medium was tested. Under the selected assay conditions 1.04 bacterial cells from the late exponential phase adhered to one Caco-2 cell, which confirms the adhesion capability of P. xylanivorans Mz5(T). The adapted adhesion test using Caco-2 cells is suitable for estimation of adhesion capability of anaerobic bacteria.

• MeSH
• bakteriální adheze fyziologie   MeSH
• buněčné kultury MeSH
• Butyrivibrio cytologie   metabolismus   MeSH
• Caco-2 buňky mikrobiologie   MeSH
• kyslík metabolismus   MeSH
• lidé MeSH
• probiotika MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kyslík 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
• bachor mikrobiologie   MeSH
• butyráty metabolismus   MeSH
• Butyrivibrio genetika   růst a vývoj   izolace a purifikace   MeSH
• Clostridium genetika   růst a vývoj   izolace a purifikace   MeSH
• DNA bakterií analýza   MeSH
• gramnegativní aerobní tyčinky a koky genetika   růst a vývoj   izolace a purifikace   MeSH
• krmivo pro zvířata MeSH
• počet mikrobiálních kolonií MeSH
• polymerázová řetězová reakce metody   MeSH
• potravní vláknina MeSH
• RNA ribozomální 16S klasifikace   genetika   MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• butyráty MeSH
• DNA bakterií MeSH
• potravní vláknina MeSH
• RNA ribozomální 16S MeSH

A strain of Aspergillus giganteus cultivated in a medium with xylan produced two xylanases (xylanase I and II) which were purified to homogeneity. Their molar mass, estimated by SDS-PAGE, were 21 and 24 kDa, respectively. Both enzymes are glycoproteins with 50 degrees C temperature optimum; optimum pH was 6.0-6.5 for xylanase I and 6.0 for xylanase II. At 50 degrees C xylanase I exhibited higher thermostability than xylanase II. Hg2+, Cu2+ and SDS were strong inhibitors, 1,4-dithiothreitol stimulated the reaction of both enzymes. Both xylanases are xylan-specific; kinetic parameters indicated higher efficiency in the hydrolysis of oat spelts xylan. In hydrolysis of this substrate, xylotriose, xylotetraose and larger xylooligosaccharides were released and hence the enzymes were classified as endoxylanases.

• MeSH
• aktivátory enzymů analýza   farmakologie   MeSH
• Aspergillus enzymologie   růst a vývoj   MeSH
• dextrany MeSH
• dithiothreitol MeSH
• dodecylsíran sodný MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• endo-1,4-beta-xylanasy chemie   izolace a purifikace   metabolismus   MeSH
• frakční precipitace MeSH
• fungální proteiny chemie   izolace a purifikace   metabolismus   MeSH
• gelová chromatografie MeSH
• glykoproteiny chemie   izolace a purifikace   metabolismus   MeSH
• inhibitory enzymů analýza   farmakologie   MeSH
• kinetika MeSH
• koncentrace vodíkových iontů MeSH
• kultivační média chemie   MeSH
• měď analýza   farmakologie   MeSH
• molekulová hmotnost MeSH
• síran amonný chemie   MeSH
• sloučeniny rtuti analýza   farmakologie   MeSH
• stabilita enzymů MeSH
• substrátová specifita MeSH
• teplota MeSH
• xylany metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktivátory enzymů MeSH
• dextrany MeSH
• dithiothreitol MeSH
• dodecylsíran sodný MeSH
• endo-1,4-beta-xylanasy MeSH
• fungální proteiny MeSH
• glykoproteiny MeSH
• inhibitory enzymů MeSH
• kultivační média MeSH
• měď MeSH
• sephadex MeSH Prohlížeč
• síran amonný MeSH
• sloučeniny rtuti MeSH
• xylany MeSH