Veterinary Archives, Vol. 94 No. 1, 2024.
Original scientific paper
https://doi.org/10.24099/vet.arhiv.1935
Identification of protein expression changes in milk in experimental bovine mastitis using difference gel electrophoresis
Funmilola Clara Thomas
orcid.org/0000-0002-1052-222X
; Department of Veterinary Physiology and Biochemistry, College of Veterinary Medicine, Federal University of Agriculture, Abeokuta, Nigeria; Institute of Biodiversity Animal Health and Comparative Medicine, University of Glasgow UK
Eyitayo Solomon Ajibola
; Department of Veterinary Physiology and Biochemistry, College of Veterinary Medicine, Federal University of Agriculture, Abeokuta, Nigeria
Allan Scott
; Institute of Cardiovascular and Medical Sciences, University of Glasgow, United Kingdom
Richardo Tassi
; Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, United Kingdom
Andre Marcos Santana
; Departamento de Medicina Veterinária Laboratório de Patologia Clínica Veterinária, Universidade Estadual de Maringá, Brazil.
Tom Nathan McNeilly
; Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, United Kingdom
Ruth Zadoks
; Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, United Kingdom
Hayley Haining
; School of Veterinary Medicine, University of Glasgow, United Kingdom
Richard Burchmore
; Institute of Infection, Immunity and Inflammation, Garscube Campus, University of Glasgow, United Kingdom
Peter David Eckersall
; Institute of Biodiversity Animal Health and Comparative Medicine, University of Glasgow UK
Abstract
In order to identify the extent of protein changes in milk during mastitis as a guide to detecting markers for prompt management of the disease, milk from cows in which clinical mastitis was experimentally induced were subjected to difference gel electrophoresis (DiGE) analysis. Pooled samples from 6 udders (from 6 cows) were analysed at selected time points: 0, 81 and 312 hours post-challenge with Streptococcus uberis mastitis. These corresponded to samples from the pre-infection, peak and resolution phases of the mastitis challenge. After the preliminary sample preparation, concentration and pooling steps, samples were labelled with CyDyes (Cydye 2, 3 and 5), after which isoelectric focusing and gel electrophoresis were carried out respectively. DiGE gels were subsequently scanned and ImageQuant, ImageJ and DeCyder™ 2D (version 7.0) softwares were used to crop, obtain jpeg images and carry out 2-D differential analysis and processing of the images, respectively. Biological variation analysis (BVA) software (GE Healthcare Life Sciences, Buckinghamshire, UK) was also used to analyse the gels and create gel to gel matching of spots (qualitatively and quantitatively) within the three gels produced. Overall, a total of 521 protein spots were identified as significantly differentially expressed (qualitatively or quantitatively) in mastitis milk during the course of the intramammary infection. This demonstrates the large repertoire of protein biomarker candidates available, revealed through this technique. Further studies are required to elucidate the merits and demerits of these changing proteins in order to identify the one most suitable for clinical application in mastitis diagnosis.
Keywords
mastitis; milk; proteins; Streptococcus uberis; difference gel electrophoresis
Hrčak ID:
315798
URI
Publication date:
9.2.2024.
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