ORIGINAL PAPER
Detecting meat-and-bone meal in ruminant’s feeds
by species-specific PCR
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1
Department of Animal Science, National I-Lan University,
#1, Sheng-Lung Rd, I-Lan 260, Taiwan
2
Department of Life Science, National Kao-Hsiung University,
Kao-Hsiung, Taiwan
3
Department of Animal Science, National Taiwan University,
Taipei, Taiwan
Publication date: 2003-10-28
Corresponding author
T. Y. Kuo
Department of Animal Science, National I-Lan University,
#1, Sheng-Lung Rd, I-Lan 260, Taiwan
J. Anim. Feed Sci. 2003;12(4):849-858
KEYWORDS
ABSTRACT
Bovine spongiform encephalopathy, first identified in the UK in 1986, may have arisen from
feeding scrapie infected meat-and-bone meal (MBM) to cattle that was produced under sub-optimal
conditions. For public health and disease prevention reasons, banning the application of MBM in ruminant’s feeds was necessary. This study utilized a polymerase chain reaction (PCR) assay to identify
bovine, porcine, ovine, and chicken MBM in animal diets. Bone fragments from feed samples were
separated by chloroform separation. A nucleospin column was used for DNA extraction. Four pairs of
primers that targeted at highly conserved regions of mitochondrial DNA (mtDNA) were employed in
a PCR procedure to detect bovine, porcine, ovine and chicken DNA sequences. These gene fragments
at the targeting region for the four species were 271, 225, 212 and 266 bps in size, respectively.
In order to confirm the specific amplification of expected products from MBM for each species,
the PCR products were digested with restriction enzymes HphI, MnlI, SspI and HindIII, and different length polymorphisms were observed. In the sensitivity assays, concentration as low as 1% of
MBM in animal feeds could be detected by the PCR procedure. Based on our prevalence survey for
commercial MBM in Taiwan, it predominantly consisted of bovine or porcine origins. This molecular
approach provides a quick, highly reproducible and sensitive method to detect the MBM in feeds.
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