No abstract

Aims: To examine the diversity of protozoa in the rumen contents of cow. Methods and Results: Protozoa that inhabit the rumen were detected by PCR using protozoan-specific primers. Libraries of protozoan rDNA sequences were constructed from rumen fluid, solid tissues and epithelium. Twentythree clones isolated from rumen fluid fell into two genera identified as Entodinium (69AE6% of clones) and Epidinium (31AE4% of clones). Of the clones isolated from rumen fluid, a moderate number were unidentifiable (30AE4%). Conclusions: The predominant protozoan genus identified in the whole rumen belonged to the Entodinium group (81AE1%). Protozoa were not detected in the rumen epithelium. Significance and Impact of the Study: These findings suggest that rumen fluid and solid tissues contain different protozoan populations that may play specific roles in rumen function. Quantitative PCR techniques and a more specific set of phylogenetic probes that distinguish between protozoan species are needed to determine the significance of newly identified groups and to determine the distribution of identified protozoan clusters in rumen microbial communities.

No abstract

The composition of yeast communities in the rumen of cattle was investigated using comparative DNA sequence analysis of yeast 26S rDNA genes. 26S rDNA libraries were constructed from rumen fluid (FF), rumen solid (FS) and rumen epithelium (FE). A total of 97 clones, containing a partial 26S rDNA sequence of 0·6 kb length, were sequenced and subjected to an on-line similarity search.The 41 FF clones could be divided into five classes. The largest class was affiliated with Pezizomycotina class (85·4% of clones), and the remaining classes were related with the Urediniomycotina (2·4%), Hymenomycetes (4·9%), Ustilaginomycetes (4·9%) and Saccharomycotina (2·4%) classes. The 26 FE clones could be divided into three classes and the Saccharomycetes class (92·4% of clones) was the largest group. The remaining classes were related with either Pezizomycotina (3·8%) or Ustilaginomycetes (3·8%). The 30 FS clones were all affiliated with Saccharomycotina. Saccharomycotina were predominant in rumen epithelium and rumen solid while Pezizomycotina were predominant in rumen fluid. Yeast belonging to the Saccharomycotina class was predominant in the rumen as a whole (57%). One clone (FF34) had less than 90% similarity to any sequence in the database and was thus apparently unrelated to any previously described yeast.

Pectobacterium chrysanthemi PY35 secretes the endoglucanase Cel5Z, an enzyme of the glycoside hydrolase family 5. Cel5Z is a 426 amino acid, signal peptide (SP)-containing protein composed of two domains: a large N-terminal catalytic domain (CD; 291 amino acids) and a small C-terminal cellulose binding domain (CBD; 62 amino acids). These two domains are separated by a 30 amino acid linker region (LR). A truncated cel5Z gene was constructed with the addition of a nonsense mutation that removes the C-terminal region of the protein. A truncated Cel5Z protein, consisting of 280 amino acid residues, functioned as a mature enzyme despite the absence of the SP, 11 amino acid CD, LR, and CBD region. In fact, this truncated Cel5Z protein showed an enzymatic activity 80% higher than that of full-length Cel5Z. However, cellulase activity was undetectable in mature Cel5Z proteins truncated to less than 280 amino acids.