Quantification of protozoa populations in rumen fluid as determined by manual counting and real time PCR | Cuantificación de protozoos en el fluido ruminal mediante recuento en cámara y PCR en tiempo real
2011
Saro, C., Universidad de León (España). Dept. de Producción Animal | Ranilla, M.J., Universidad de León (España). Dept. de Producción Animal | Carro, M.D., Universidad de León (España). Dept. de Producción Animal
Four cannulated merino sheep were used to evaluate the real time PCR (qPCR) method to quantify protozoa populations in rumen fluid. Two diets were formulated with 70:30 forage:concentrate ratio and alfalfa hay (AL) or grass hay (GR) as forage. Rumen fluid was sampled before feeding (0 h) and 4 and 8 h after feeding. Protozoa data from qPCR were expressed relative to absolute quantification of total bacteria population and were compared with results from manual counting of protozoa in a microscopic chamber. Results from both methods were different. In manual counting a decrease in protozoa number was observed at 4 and 8 hours post feeding (P<0.05), whereas qPCR results did not detect any change (P<0.05) between sampling times. Protozoa population was higher (P<0.015) for AL compared with GR diet with manual counting, whereas diet GR promoted a higher (P<0.010) protozoa population than AL diet with the qPCR method. Differences between methods may have been due to the fact that the results were expressed in relation to total bacteria quantification, and bacteria concentrations varied between sampling times.
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