The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.
Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen: Real-time PCR assay of the rumen anaerobic fungal population
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.
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Altschul, S. F., Gish, W., Miller, W., Myers, E. W., & Lipman, D. J. (1990) · Journal of Molecular Biology · doi:10.1006/jmbi.1990.9999 · Cited by 98,669
Klieve, A. V., Hennessy, D., Ouwerkerk, D., Forster, R. J., Mackie, R. I., & Attwood, G. T. (2003) · Journal of Applied Microbiology · doi:10.1046/j.1365-2672.2003.02024.x · Cited by 248
Distribution of anaerobic fungi in the digestive tract of cattle and their survival in faeces
Davies, D. R., Theodorou, M. K., Lawrence, M. I. G., & Trinci, A. P. J. (1993) · Journal of General Microbiology · doi:10.1099/00221287-139-6-1395 · Cited by 117
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“The program for detection of the total bacterial population [26] consisted of an initial 4 min, 95°C hot start followed by 35 cycles of 95° C for 20 s, 60°C for 20 s and 72°C for 45 s with fluorescent acquisition after each cycle.”
Metadata
- Authors
- Denman, S. E., & McSweeney, C. S.
- Venue
- FEMS Microbiology Ecology
- Publisher
- Oxford University Press (OUP)
- Published
- 2006-12
- Volume
- 58
- Issue
- 3
- Pages
- 572-582
- ISSN
- 0168-6496 (ISSN-L)
- 1574-6941
- OpenAlex
- W1817506593
- Cited by
- 754
- References
- 3
- FWCI
- 23.21
- Type
- journal article
- Language
- en
- Open access
- closed
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- Denman, S. E., & McSweeney, C. S. (2006). Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen: Real-time PCR assay of the rumen anaerobic fungal population. FEMS Microbiology Ecology. https://doi.org/10.1111/j.1574-6941.2006.00190.x
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