An rRNA-based analysis for evaluating the effect of heat stress on the rumen microbial composition of Holstein heifers.

We performed a set of heifer feeding trials to investigate the effect of heat and humidity stresses on the rumen bacterial molecular diversity of Holstein heifers (Tajima K, Nonaka I, Higuchi K, Takusari N, Kurihara M, Takenaka A, et al. Anaerobe 2007;13:57-64). To further characterize the response of the microbial community to the physiological changes caused by the stresses, we evaluated changes in the ruminal bacterial community composition in the same trials by applying an RNA-based method (sequence-specific small-subunit (SSU) rRNA cleavage method), which was optimized for a comprehensive description of the predominant bacterial groups inhabiting the rumen. Four Holstein heifers were kept at three temperatures (20 degrees C, 28 degrees C, 33 degrees C) in a climatic chamber for two weeks each, and rumen fluid samples were obtained on the last day of each temperature experiment. For quantitative detection, we applied a set of 15 oligonucleotide probes, including those targeting taxa comprised of uncultured rumen bacteria (URB) belonging to phylum Firmicutes, to the RNAs extracted from the fluid samples. The relative populations of the Clostridium coccoides-Eubacterium rectale group, and the genus Streptococcus increased, and that of the genus Fibrobacter decreased in response to increasing temperature both in the first (nine months old, 80% relative humidity) and second (15 months old, 60% relative humidity) experiments. In addition, the population of a defined URB group was higher at 33 degrees C than at 20 degrees C in the second trial, whereas one of the other URB groups showed a decreasing trend with the temperature rise. These results indicate that the exposure to heat affects the population levels of specific bacterial groups in the ruminal microbial community. 2009 Elsevier Ltd. All rights reserved.