Skip to main content
ARS Home » Midwest Area » Bowling Green, Kentucky » Food Animal Environmental Systems Research » Research » Publications at this Location » Publication #399113

Research Project: Developing Agronomically and Environmentally Beneficial Management Practices to Increase the Sustainability and Safety of Animal Manure Utilization

Location: Food Animal Environmental Systems Research

Title: Liquid chromatography mass spectrometry (LCMS) method development for determination of ß-defensins in bovine milk

item OGBEBOR, ELISHA - Western Kentucky University
item CONTE, ERIC - Western Kentucky University
item KERRO DEGO, OUDESS - University Of Tennessee
item Agga, Getahun

Submitted to: American Chemical Society Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: 12/23/2022
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Bovine mastitis is caused by a wide range of pathogens which results in a substantial economic loss for the dairy cattle livestock industry. ß-defensins are a part of the innate immune system and act as the first line of defense against mastitis in bovine. ß-defensins (~6-10kDa) are antimicrobial peptides that contain about 28 to 67 amino acids, the presence of six cysteine residues results in three disulfide bonds formed and a ß-sheet structure. A few ß-defensins; TAP (Tracheal antimicrobial peptide), LAP (Lingual antimicrobial peptide), DEFB1 (ß-defensin 1 peptide), DEFB3 (ß-defensin 3 peptide), DEFB4 (ß-defensin 4 peptide), DEFB5 (ß-defensin 5 peptide), DEFB10 (ß-defensin 10 peptide), and DEFB103 (ß-defensin 103 peptide) have been identified in cows based on site of occurrence and time of expression in tissue, but not in milk. A Solid Phase Extraction (SPE) and Liquid Chromatography Mass Spectrometry (LCMS) method is developed to identify ß-defensins in bovine milk of healthy and mastitis-infected cows. The LC-MS data for infected bovine milk was compared with the data obtained from the healthy sample. The peptides were separated on a Phenomenex bioZen Intact XB-C8 column (150 x 2.1 mm, 3.6 µm) on a Thermo Scientific HPLC system using an acetonitrile:H2O in 0.1% formic acid mobile phase gradient coupled with the TSQ Thermo Fortis mass spectrometer. A scan range from 100 –2000 m/z in the positive ion mode is used for the acquisition. Other antimicrobial ß-defensin peptides are expected in mastitis-infected bovine milk and will be explored.