Distinguishing Natural Infections of the Bovine Mammary Gland by <i>Staphylococcus</i> from <i>Streptococcus</i> spp. Using Quantitative Milk Proteomics
Dina Rešetar Maslov,
Funmilola Clara Thomas,
Anđelo Beletić,
Josipa Kuleš,
Ivana Rubić,
Miroslav Benić,
Goran Bačić,
Nino Maćešić,
Vida Eraghi,
Vladimir Farkaš,
Tihana Lenac Roviš,
Berislav Lisnić,
Damir Žubčić,
Dalibor Potočnjak,
Vladimir Mrljak
Affiliations
Dina Rešetar Maslov
Laboratory of Proteomics, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Funmilola Clara Thomas
Laboratory of Proteomics, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Anđelo Beletić
Laboratory of Proteomics, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Josipa Kuleš
Department of Chemistry and Biochemistry, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Ivana Rubić
Laboratory of Proteomics, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Miroslav Benić
Department of Bacteriology and Parasitology, Croatian Veterinary Institute, Savska Cesta, 143, 10000 Zagreb, Croatia
Goran Bačić
Reproduction and Obstetrics Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Nino Maćešić
Reproduction and Obstetrics Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Vida Eraghi
Laboratory of Proteomics, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Vladimir Farkaš
Laboratory of Proteomics, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Tihana Lenac Roviš
Center for Proteomics University of Rijeka, Faculty of Medicine, Brace Branchetta 20, 51000 Rijeka, Croatia
Berislav Lisnić
Center for Proteomics University of Rijeka, Faculty of Medicine, Brace Branchetta 20, 51000 Rijeka, Croatia
Damir Žubčić
Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Dalibor Potočnjak
Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Vladimir Mrljak
Laboratory of Proteomics, Internal Diseases Clinic, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova Street 55, 10000 Zagreb, Croatia
Bovine mastitis is the most frequent disease on dairy farms, which leads to a decrease in the health welfare of the animals and great economic losses. This study was aimed at determining the quantitative variations in the milk proteome caused by natural infection by Staphylococcus and Streptococcus species in order to gain further understanding of any discrepancies in pathophysiology and host immune responses, independent of the mastitis level. After identification of Staphylococcus (N = 51) and Streptococcus (N = 67) spp., tandem mass tag (TMT)-labeled quantitative proteomic and liquid chromatography-mass spectrometry (LC-MS/MS) techniques on a modular Ultimate 3000 RSLCnano system coupled to a Q Exactive Plus was applied on aseptically sampled milk from Holstein cows. Proteome Discoverer was used for protein identification and quantitation through the SEQUEST algorithm. Statistical analysis employing R was used to identify differentially abundant proteins between the groups. Protein classes, functions and functional-association networks were determined using the PANTHER and STRING tools and pathway over-representation using the REACTOME. In total, 156 master bovine proteins were identified (two unique peptides, p p < 0.05 and FDR < 0.5). The most discriminatory proteins per group were odorant-binding protein (higher in staphylococci) and fibrinogen beta chain protein (higher in streptococci). The receiver operating characteristic (ROC) curve showed that protein kinase C-binding protein NELL2, thrombospondin-1, and complement factor I have diagnostic potential for differentiating staphylococci and streptococci intramammary infection and inflammation. Improved understanding of the host response mechanisms and recognition of potential biomarkers of specific-pathogen mastitis, which may aid prompt diagnosis for control implementation, are potential benefits of this study.
