Abstract:

【Objective】 Using Real-time PCR (RT-PCR)assays to quantify the microbes associated with particular phases of rumen contents. 【Method】 Whole rumen contents were sampled from three Mongolian sheep and partitioned into solid- and liquid-associated microorganisms. 【Result】 On the basis of 16S or 18S rRNA gene copy number analysis, the number of general bacteria, general anaerobic fungi and three representative ruminal cellulolytic bacteria (Ruminococcus albus, Ruminococcus flavefaciens, Fibrobacter succinogenes) associated with rumen solid content were 7.22-fold (P＜0.05), 56.85-fold (P＜0.01), 2.69-fold (P＜0.05), 4.19-fold (P＜0.01) and 7.59-fold (P＜0.01) as those associated with rumen liquid content. The abundance of F. succinogenes in the solid phase was 0.55%, which was higher than that of R. flavefaciens (0.53%) and R. albus (0.09%). However in the liquid phase R. flavefaciens accounted for 0.91% of the bacterial rRNA gene copies, which was more abundant than F. succinogenes (0.52%) and R. albus (0.24%). 【Conclusion】 RT-PCR method could accurately detect the distribution and abundance of microorganisms associated with particular phase of rumen content.

Key words: Real-time PCR, Mongolia sheep, rumen, microorganisms, distribution