The rumen is one of the most powerful fibrolytic fermentation systems known. Gene expression analyses, such as reverse transcription PCR (RT-PCR), microarrays, and metatranscriptomics, are techniques that could significantly expand our understanding of this ecosystem. The ability to isolate and stabilize representative RNA samples is critical to obtaining reliable results with these procedures. In this study, we successfully isolated high-quality total RNA from the solid phase of ruminal contents by using an improved RNA extraction method. This method is based on liquid nitrogen grinding of whole ruminal solids without microbial detachment and acid guanidinium - phenol - chloroform extraction combined with column purification. Yields of total RNA were as high as 150 µg per g of fresh ruminal content. The typical large subunit/small subunit rRNA ratio ranged from 1.8 to 2.0 with an RNA integrity number (Agilent Technologies) greater than 8.5. By eliminating the detachment step, the resulting RNA was more representative of the complete ecosystem. Our improved method removed a major barrier limiting analysis of rumen microbial function from a gene expression perspective. The polyA-tailed eukaryotic mRNAs obtained have successfully been applied to next-generation sequencing, and metatranscriptomic analysis of the solid fraction of rumen contents revealed abundant sequences related to rumen fungi.

The importance of factors such as cultural conditions e.g. organism background, inoculum density and glucose concentration are explored and their effects on the experimental recording discussed. The relevance of these findings to microbial microcalorimetry in general and to microcalorimetric identification of organisms in particular is described. The conclusions drawn are (i) that microbial identification by microcalorimetry is not possible and (ii) that great caution must be exercised in interpreting metabolic data derived from microcalorimetric studies.

A flow microcalorimetric procedure has been developed for the characterization of representative strains of Saccharomyces cerevisiae and Sacch. uvarum (carlsbergensis) used in the food industry. Growth in a chemically defined medium containing single and mixed sugars gave reproducible and characteristic power‐time curves. The technique is proposed as a rapid alternative method for the characterization of yeast strains selected for commercial use.

SUMMARYThe effect of hypochlorite concentration on Mycoplasma mycoides ssp. mycoides viability was tested under a variety of conditions. The experimental variables employed included chlorine-cell contact time, chlorine concentration, carrier system and organic loading. Initial populations of 106 c.f.u./ml were killed (no survivors in 1

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