Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen

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ID: 306795
2006
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Abstract
Traditional methods for enumerating and identifying microbial populations within the rumen can be time consuming and cumbersome. Methods that involve culturing and microscopy can also be inconclusive, particularly when studying anaerobic rumen fungi. A real-time PCR SYBR Green assay, using PCR primers to target total rumen fungi and the cellulolytic bacteria Ruminococcus flavefaciens and Fibrobacter succinogenes, is described, including design and validation. The DNA and crude protein contents with respect to the fungal biomass of both polycentric and monocentric fungal isolates were investigated across the fungal growth stages to aid in standard curve generation. The primer sets used were found to be target specific with no detectable cross-reactivity. Subsequently, the real-time PCR assay was employed in a study to detect these populations within cattle rumen. The anaerobic fungal target was observed to increase 3.6-fold from 0 to 12 h after feeding. The results also indicated a 5.4-fold increase in F. succinogenes target between 0 and 12 h after feeding, whereas R. flavefaciens was observed to maintain more or less consistent levels. This is the first report of a real-time PCR assay to estimate the rumen anaerobic fungal population.
Reference Key
openalex_W1817506593 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Stuart E. Denman, Christopher S. McSweeney
Journal fems microbiology ecology
Year 2006
DOI
10.1111/j.1574-6941.2006.00190.x
URL
https://doi.org/10.1111/j.1574-6941.2006.00190.x
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