[EHEC carriage in ruminants and probiotic effects].

Enterohaemorrhagic Escherichia coli (EHEC) are Shiga-Toxin producing E. coli (STEC) that cause human outbreaks which can lead to a severe illness such as haemolytic-uraemic syndrome (HUS), particularly in young children. The gastrointestinal tract of cattle and other ruminants is the principal reservoir of EHEC strains and outbreaks have been associated with direct contact with the farm environment, and with the consumption of meat, dairy products, water and fruit or vegetable contaminated with ruminant manure.

Rumen microbial and fermentation characteristics are affected differently by bacterial probiotic supplementation during induced lactic and subacute acidosis in sheep.

Background: Ruminal disbiosis induced by feeding is the cause of ruminal acidosis, a digestive disorder prevalent in high-producing ruminants. Because probiotic microorganisms can modulate the gastrointestinal microbiota, propionibacteria- and lactobacilli-based probiotics were tested for their effectiveness in preventing different forms of acidosis.

Results: Lactic acidosis, butyric and propionic subacute ruminal acidosis (SARA) were induced by feed chalenges in three groups of four wethers intraruminally dosed with wheat, corn or beet pulp.

Ruminococcus champanellensis sp. nov., a cellulose-degrading bacterium from human gut microbiota.

A strictly anaerobic, cellulolytic strain, designated 18P13(T), was isolated from a human faecal sample. Cells were Gram-positive non-motile cocci. Strain 18P13(T) was able to degrade microcrystalline cellulose but the utilization of soluble sugars was restricted to cellobiose.

The cellulose-degrading microbial community of the human gut varies according to the presence or absence of methanogens.

Cellulose-degrading microorganisms involved in the breakdown of plant cell wall material in the human gut remain rather unexplored despite their role in intestinal fermentation. Microcrystalline cellulose-degrading bacteria were previously identified in faeces of methane-excreting individuals, whereas these microorganisms were undetectable in faecal samples from non-methane excretors. This suggested that the structure and activity of the cellulose-degrading community differ in methane- and non-methane-excreting individuals.

Characterization of Xyn10A, a highly active xylanase from the human gut bacterium Bacteroides xylanisolvens XB1A.

A xylanase gene xyn10A was isolated from the human gut bacterium Bacteroides xylanisolvens XB1A and the gene product was characterized. Xyn10A is a 40-kDa xylanase composed of a glycoside hydrolase family 10 catalytic domain with a signal peptide. A recombinant His-tagged Xyn10A was produced in Escherichia coli and purified.

Experimental feed induction of ruminal lactic, propionic, or butyric acidosis in sheep.

A study was conducted to determine the feasibility to induce rumen acidosis with propionate, butyrate, or lactate as the major fermentation end products. Three rumen-cannulated Texel wethers were used in a 3 x 3 Latin square design. Each period consisted of 11 d of adaptation where wethers were daily fed at 90% of ad libitum intake a hay and wheat-based concentrate diet (4:1 ratio on a DM basis) in 2 equal portions followed by 3 d of acidosis induction.

Dietary fibre degradation and fermentation by two xylanolytic bacteria Bacteroides xylanisolvens XB1A and Roseburia intestinalis XB6B4 from the human intestine.

Aims: To characterize fibre degradation, colonization and fermentation, and xylanase activity of two xylanolytic bacteria Bacteroides xylanisolvens XB1A(T) and Roseburia intestinalis XB6B4 from the human colon.

Methods And Results: The bacteria grew well on all the substrates chosen to represent dietary fibres: wheat and corn bran, pea, cabbage and leek fibres, and also on purified xylans. Roseburia intestinalis colonized the substrates more efficiently than Bact.

Assessment of metabolic diversity within the intestinal microbiota from healthy humans using combined molecular and cultural approaches.

The human gut harbours a wide range of bacterial communities that play key roles in supplying nutrients and energy to the host through anaerobic fermentation of dietary components and host secretions. This fermentative process involves different functional groups of microorganisms linked in a trophic chain. Although the diversity of the intestinal microbiota has been studied extensively using molecular techniques, the functional aspects of this biodiversity remain mostly unexplored.

Bacteroides xylanisolvens sp. nov., a xylan-degrading bacterium isolated from human faeces.

During the course of a study on the xylan-degrading community from the human gut, six xylanolytic, Gram-negative, anaerobic rods were isolated from faecal samples. 16S rRNA gene sequence analysis showed that the isolates were closely related to each other (> or =99 % sequence similarity) and that they belonged to the genus Bacteroides. On the basis of 16S rRNA gene sequence similarity, representative strain XB1AT was most closely related to the type strains of Bacteroides ovatus (97.

Image analysis and data normalization procedures are crucial for microarray analyses.

This study was conducted with the aim of optimizing the experimental design of array experiments. We compared two image analysis and normalization procedures prior to data analysis using two experimental designs. For this, RNA samples from Charolais steers Longissimus thoracis muscle and subcutaneous adipose tissues were labeled and hybridized to a bovine 8,400 oligochip either in triplicate or in a dye-swap design.

Bacteroides cellulosilyticus sp. nov., a cellulolytic bacterium from the human gut microbial community.

A strictly anaerobic cellulolytic bacterium, strain CRE21(T), was isolated from a human faecal sample. Cells were Gram-negative non-motile rods that were about 1.7 microm in length and 0.

Characterization of the xylan-degrading microbial community from human faeces.

In humans, plant cell wall polysaccharides represent an important source of dietary fibres that are digested by gut microorganisms. Despite the extensive degradation of xylan in the colon, the population structure and the taxonomy of the predominant bacteria involved in degradation of this polysaccharide have not been extensively explored. The objective of our study was to characterize the xylanolytic microbial community from human faeces, using xylan from different botanic origins.

Optimizing rumen functions in the close-up transition period and early lactation to drive dry matter intake and energy balance in cows.

The large deficit in energy intake in relation to energy requirements during the transition and early lactation periods means that high-producing cows need energy-dense rations. High-starch diets are intensively fermented by the microbial ecosystem in the rumen, giving rise to a high production of VFAs and resulting in a drop in pH and the accumulation of lactic acid, which exacerbates the decline in pH and is considered as the major cause of rumen acidosis. This rumen dysfunction affects rumen microbes and results in less efficient digestion, thereby decreasing feed intake and exacerbating the energy deficit in the cows.

Effect of Methanobrevibacter sp MF1 inoculation on glycoside hydrolase and polysaccharide depolymerase activities, wheat straw degradation and volatile fatty acid concentrations in the rumen of gnotobiotically-reared lambs.

Four naturally born lambs were placed in sterile isolators 24 h after birth before the natural establishment of cellulolytic microorganisms and archaea methanogens. At the age of 6 weeks they were inoculated with pure cultures of the strains FD1 and 007 of Ruminococcus flavefaciens and at the age of 4 months with a pure culture of Methanobrevibacter sp. MF1.

H2 and acetate transfers during xylan fermentation between a butyrate-producing xylanolytic species and hydrogenotrophic microorganisms from the human gut.

The aim of this work was to investigate in vitro interrelationships during xylan fermentation between an H2 and butyrate-producing xylanolytic species recently isolated in our laboratory from human faeces and identified as Roseburia intestinalis and the H2-utilizing acetogen Ruminococcus hydrogenotrophicus or the methanogen Methanobrevibacter smithii. H2 transfer between M. smithii or Ru.

The Ruminococcus albus pilA1-pilA2 locus: expression and putative role of two adjacent pil genes in pilus formation and bacterial adhesion to cellulose.

Ruminococcus albus produces fimbria-like structures that are involved with the bacterium's adhesion to cellulose. The subunit protein has been identified in strain 8 (CbpC) and strain 20 (GP25) and both are type IV fimbrial (Pil) proteins. The presence of a pil locus that is organized similarly in both strains is reported here together with the results of an initial examination of a second Pil protein.

Xylanases and carboxymethylcellulases of the rumen protozoa Polyplastron multivesiculatum, Eudiplodinium maggii and Entodinium sp.

Endoglucanase and xylanase activities of three rumen protozoa, Polyplastron multivesiculatum, Eudiplodinium maggii, and Entodinium sp. were compared qualitatively by zymograms and quantitatively by measuring specific activities against different polysaccharides. A set of carboxymethylcellulases and xylanases was produced by the large ciliates whereas no band of activity was observed for Entodinium sp.

A collection of bovine cDNA probes for gene expression profiling in muscle.

Array technology has been increasingly used to monitor global gene expression patterns in various tissues and cell types. However, applications to muscle development and pathology as well as meat production in livestock species have been hampered by the lack of appropriate cDNA collections. To overcome this problem, a directed cDNA library was constructed starting from 23 muscles of meat-producing bovines to derive a collection of 3573 clones.

Interaction between H2-producing and non-H2-producing cellulolytic bacteria from the human colon.

The cellulose-degrading species recently isolated from the human colon showed diverse ability to degrade and ferment cellulose. In the present study, the nature of the inter-relation existing between one H(2)-producing cellulolytic isolate (Ruminococcus sp. nov.

Reconstruction of the evolutionary history of the LexA-binding sequence.

In recent years, the recognition sequence of the SOS repressor LexA protein has been identified for several bacterial clades, such as the Gram-positive, green non-sulfur bacteria and Cyanobacteria phyla, or the 'Alphaproteobacteria', 'Deltaproteobacteria' and 'Gammaproteobacteria' classes. Nevertheless, the evolutionary relationship among these sequences and the proteins that recognize them has not been analysed. Fibrobacter succinogenes is an anaerobic Gram-negative bacterium that branched from a common bacterial ancestor immediately before the Proteobacteria phylum.

Small intestine and liver microsomal triacylglycerol transfer protein in the bovine and rat: effects of dietary coconut oil.

The bovine liver is characterized by a chronic low capacity to secrete triacylglycerols (TAG). In situations favoring their hepatic synthesis, such as coconut oil feeding, TAG accumulate, leading to a lipid infiltration in the liver of preruminant calves. To assess the possible role of the microsomal TAG transfer protein (MTP) in this phenomenon and to put into evidence a tissue-specific regulation in the bovine species, we compared by Western blot the content in both MTP subunits in the liver and in different portions of the small intestine in preruminant calves and in growing rats receiving coconut oil or beef tallow as the sole source of fat in the diet.

Linoleate supplementation in steers modifies lipid composition of plasma lipoproteins but does not alter their fluidity.

The health value for man of lipids in bovine muscles can be improved by the addition of PUFA to the animals' diets, but such treatments can modify fluidity of plasma lipoproteins and therefore their metabolic functions. The aim of the present study was to analyse whether changes in chemical composition of lipoproteins in steers fed sunflower oil-rich diets altered lipoprotein fluidity, measured by fluorescence polarization and electron spin resonance. LDL, light HDL and heavy HDL fractions were isolated by ultracentrifugation from plasma of eighteen crossbred Charolais x Salers steers.

Two-step freezing procedure for cryopreservation of rumen ciliates, an effective tool for creation of a frozen rumen protozoa bank.

The present study aimed at the long-term storage of rumen protozoa as living cells in liquid nitrogen. The two-step or interrupted slow freezing procedure was used to cryopreserve six of the dominant species of rumen ciliates isolated from monofaunated animals, Dasytricha ruminantium, Entodinium caudatum, Epidinium ecaudatum caudatum, Eudiplodinium maggii, Isotricha prostoma, and Polyplastron multivesiculatum. We optimized the first step in the interrupted slow freezing procedure, from the extracellular ice nucleation temperature to the holding temperature, and studied the effects of the cooling rates on survival.

Adhesion to cellulose of the Gram-positive bacterium Ruminococcus albus involves type IV pili.

This study was aimed at characterizing a cell-surface 25 kDa glycoprotein (GP25) that was previously shown to be underproduced by a spontaneous adhesion-defective mutant D5 of Ruminococcus albus 20. An antiserum against wild-type strain 20 was adsorbed with the mutant D5 to enrich it in antibodies 'specific' to adhesion structures of R. albus 20.

Interspecies H2 transfer in cellulose degradation between fibrolytic bacteria and H2-utilizing microorganisms from the human colon.

Interspecies H2 transfer between two newly isolated fibrolytic strains (18P13 and 18P16) and H2-utilizing methanogen or acetogen from the human colon was investigated during in vitro cellulose degradation. Both H2-consuming microorganisms utilized efficiently H2 produced from cellulose fermentation by the fibrolytic species. H2 utilization by Methanobrevibacter smithii did not change the metabolism and the cellulolytic activity of strain 18P16 whereas it induced a metabolic shift in strain 18P13.