Italian Journal of Animal Science (Feb 2010)

Sampling methods for rumen microbial counts by Real-Time PCR techniques

  • S. Puppo,
  • B. Ferri,
  • F. Grandoni,
  • A. Chiariotti

DOI
https://doi.org/10.4081/ijas.2007.s2.788
Journal volume & issue
Vol. 6, no. 2s
pp. 788 – 791

Abstract

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Fresh rumen samples were withdrawn from 4 cannulated buffalo females fed a fibrous diets in order to quantify bacteria concentration in the rumen by Real-Time PCR techniques. To obtain DNA of a good quality from whole rumen fluid, eight (M1-M8) different pre-filtration methods (cheese cloths, glass-fibre and nylon filter) in combination with various centrifugation speeds (1000, 5000 and 14,000 rpm) were tested. Genomic DNA extraction was performed either on fresh or frozen samples (-20°C). The quantitative bacteria analysis was realized according to Real-Time PCR procedure for Butyrivibrio fibrisolvens reported in literature. M5 resulted the best sampling procedure allowing to obtain a suitable genomic DNA. No differences were revealed between fresh and frozen samples.

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