title: Mastitomics, the integrated omics of bovine milk in an experimental model of Streptococcus uberis mastitis: 2. Label-free relative quantitative proteomics.
abstract: Mastitis, inflammation of the mammary gland, is the most common and costly disease of dairy cattle in the western world. It is primarily caused by bacteria, with Streptococcus uberis as one of the most prevalent causative agents. To characterize the proteome during Streptococcus uberis mastitis, an experimentally induced model of intramammary infection was used. Milk whey samples obtained from 6 cows at 6 time points were processed using label-free relative quantitative proteomics. This proteomic analysis complements clinical, bacteriological and immunological studies as well as peptidomic and metabolomic analysis of the same challenge model. A total of 2552 non-redundant bovine peptides were identified, and from these, 570 bovine proteins were quantified. Hierarchical cluster analysis and principal component analysis showed clear clustering of results by stage of infection, with similarities between pre-infection and resolution stages (0 and 312 h post challenge), early infection stages (36 and 42 h post challenge) and late infection stages (57 and 81 h post challenge). Ingenuity pathway analysis identified upregulation of acute phase protein pathways over the course of infection, with dominance of different acute phase proteins at different time points based on differential expression analysis. Antimicrobial peptides, notably cathelicidins and peptidoglycan recognition protein, were upregulated at all time points post challenge and peaked at 57 h, which coincided with 10 000-fold decrease in average bacterial counts. The integration of clinical, bacteriological, immunological and quantitative proteomics and other-omic data provides a more detailed systems level view of the host response to mastitis than has been achieved previously.
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Jarret Building, Bearsden Road, Glasgow, G61 1QH, UK. ruth.zadoks@glasgow.ac.uk and Glasgow Polyomics, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.
authors.forename: Manikhandan
authors.initials: M
authors.lastname: Mudaliar
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, UK.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Tassi
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Jarret Building, Bearsden Road, Glasgow, G61 1QH, UK. ruth.zadoks@glasgow.ac.uk.
authors.forename: Funmilola C
authors.initials: FC
authors.lastname: Thomas
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, UK.
authors.forename: Tom N
authors.initials: TN
authors.lastname: McNeilly
authors.affiliation: Glasgow Polyomics, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.
authors.forename: Stefan K
authors.initials: SK
authors.lastname: Weidt
authors.affiliation: School of Veterinary Medicine, University of Glasgow, Glasgow, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: McLaughlin
authors.affiliation: Glasgow Polyomics, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.
authors.forename: David
authors.initials: D
authors.lastname: Wilson
authors.affiliation: Glasgow Polyomics, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK and Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Burchmore
authors.affiliation: Glasgow Polyomics, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK and Institute of Molecular Cell and Systems Biology, University of Glasgow, Glasgow, UK.
authors.forename: Pawel
authors.initials: P
authors.lastname: Herzyk
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Jarret Building, Bearsden Road, Glasgow, G61 1QH, UK. ruth.zadoks@glasgow.ac.uk.
authors.forename: P David
authors.initials: PD
authors.lastname: Eckersall
authors.affiliation: Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Jarret Building, Bearsden Road, Glasgow, G61 1QH, UK. ruth.zadoks@glasgow.ac.uk and Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, UK.
authors.forename: Ruth N
authors.initials: RN
authors.lastname: Zadoks
mesh_terms: D000818:Animals; D015415:Biomarkers; D002417:Cattle; D002853:Chromatography, Liquid; D016000:Cluster Analysis; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D008414:Mastitis, Bovine / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D008892:Milk / Q000378:metabolism*; D008894:Milk Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D025341:Principal Component Analysis; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction; D013290:Streptococcal Infections / Q000662:veterinary*; D013291:Streptococcus; D053719:Tandem Mass Spectrometry
chemical_list: D015415:Biomarkers; D008894:Milk Proteins; D010455:Peptides