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Updated: Aug 20, 2026
Use of Bisection to Reduce Mitochondrial DNA in the Bovine Oocyte
Published on: July 6, 2022
PCR detection of bovine mitochondrial DNA derived from meat and bone meal in feed
Atsushi Toyoda1, Mitsuru Nakajo, Hiroyuki Kawachi
1College of Agriculture, Ibaraki University, Ibaraki 300-0393, Japan. atoyoda@mx.ibaraki.ac.jp
Abstract:
Because bovine meat and bone meal (MBM) is thought to be a major source of bovine spongiform encephalopathy, we developed a PCR-based method for detection of bovine MBM in animal feed. We isolated bone particles from feed containing bovine MBM using a separation technique based on specific gravity and then washed bone particles with sodium hypochlorite solution and an EDTA-proteinase K solution. The mitochondrial DNA was extracted from bone particles and amplified using PCR with cattle-specific primers. Bovine DNA was not detected in a milk replacer containing dried skim milk and dried whey, but bovine DNA was detected in the milk replacer that was mixed with bovine MBM. Other cattle-derived materials in feeds did not interfere with the selective detection of bovine MBM. This method allowed detection of bovine mitochondrial DNA in feed with 0.1% added bovine MBM. When the treatment with sodium hypochlorite was excluded, bovine DNA derived from MBM could not be distinguished from bovine DNA derived from other bovine materials. However, the exclusion of this treatment improved the detection limit of bovine MBM in feed. This method appears suitable for the selective detection of bovine MBM in feed.
Insights
A new PCR method detects bovine meat and bone meal (MBM) in animal feed, crucial for preventing bovine spongiform encephalopathy. This technique accurately identifies even small amounts of MBM, enhancing feed safety.
Area of Science:
- Veterinary Science
- Molecular Biology
- Food Safety
Background:
- Bovine meat and bone meal (MBM) is a suspected source of bovine spongiform encephalopathy (BSE).
- Accurate detection of MBM in animal feed is essential for preventing disease transmission.
- Existing methods may lack specificity or sensitivity for MBM detection.
Purpose of the Study:
- To develop a selective and sensitive PCR-based method for detecting bovine MBM in animal feed.
- To differentiate bovine MBM from other bovine-derived materials in feed.
- To ensure the safety of animal feed by identifying potential BSE sources.
Main Methods:
- Isolation of bone particles from feed using specific gravity separation.
- Washing bone particles with sodium hypochlorite and EDTA-proteinase K solutions.
- Extraction of mitochondrial DNA and amplification using cattle-specific PCR primers.
Main Results:
- Bovine DNA was detected in feed mixed with bovine MBM, but not in milk replacer without MBM.
- The method selectively detected bovine MBM, unaffected by other cattle-derived materials.
- Detection of bovine mitochondrial DNA was possible down to 0.1% added bovine MBM.
- Excluding sodium hypochlorite treatment improved detection limits but reduced selectivity.
Conclusions:
- The developed PCR method is suitable for the selective detection of bovine MBM in animal feed.
- The method contributes to enhanced feed safety and BSE prevention strategies.
- Further optimization may balance sensitivity and selectivity for practical application.
