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Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics

SIMPLE SUMMARY: Detection of new diagnostic markers would greatly influence the institution of preventive (control) protocols in bovine mastitis. Using the quantitative proteomics workflows and statistics for milk analysis, we identified that protein kinase C-binding protein NELL2, thrombospondin-1,...

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Autores principales: Rešetar Maslov, Dina, Thomas, Funmilola Clara, Beletić, Anđelo, Kuleš, Josipa, Rubić, Ivana, Benić, Miroslav, Bačić, Goran, Maćešić, Nino, Eraghi, Vida, Farkaš, Vladimir, Lenac Roviš, Tihana, Lisnić, Berislav, Žubčić, Damir, Potočnjak, Dalibor, Mrljak, Vladimir
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10252062/
https://www.ncbi.nlm.nih.gov/pubmed/37889706
http://dx.doi.org/10.3390/ani13111829
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author Rešetar Maslov, Dina
Thomas, Funmilola Clara
Beletić, Anđelo
Kuleš, Josipa
Rubić, Ivana
Benić, Miroslav
Bačić, Goran
Maćešić, Nino
Eraghi, Vida
Farkaš, Vladimir
Lenac Roviš, Tihana
Lisnić, Berislav
Žubčić, Damir
Potočnjak, Dalibor
Mrljak, Vladimir
author_facet Rešetar Maslov, Dina
Thomas, Funmilola Clara
Beletić, Anđelo
Kuleš, Josipa
Rubić, Ivana
Benić, Miroslav
Bačić, Goran
Maćešić, Nino
Eraghi, Vida
Farkaš, Vladimir
Lenac Roviš, Tihana
Lisnić, Berislav
Žubčić, Damir
Potočnjak, Dalibor
Mrljak, Vladimir
author_sort Rešetar Maslov, Dina
collection PubMed
description SIMPLE SUMMARY: Detection of new diagnostic markers would greatly influence the institution of preventive (control) protocols in bovine mastitis. Using the quantitative proteomics workflows and statistics for milk analysis, we identified that protein kinase C-binding protein NELL2, thrombospondin-1, and complement factor I have diagnostic potential for differentiating staphylococci and streptococci intramammary natural infection and inflammation, and subsequently gained a deeper understanding of the immunopathology of mastitis-related infection and inflammation for various etiological agents. ABSTRACT: 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 < 0.05 and FDR < 0.001), and 20 proteins showed significantly discrepant abundance between the genera (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.
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spelling pubmed-102520622023-06-10 Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics Rešetar Maslov, Dina Thomas, Funmilola Clara Beletić, Anđelo Kuleš, Josipa Rubić, Ivana Benić, Miroslav Bačić, Goran Maćešić, Nino Eraghi, Vida Farkaš, Vladimir Lenac Roviš, Tihana Lisnić, Berislav Žubčić, Damir Potočnjak, Dalibor Mrljak, Vladimir Animals (Basel) Article SIMPLE SUMMARY: Detection of new diagnostic markers would greatly influence the institution of preventive (control) protocols in bovine mastitis. Using the quantitative proteomics workflows and statistics for milk analysis, we identified that protein kinase C-binding protein NELL2, thrombospondin-1, and complement factor I have diagnostic potential for differentiating staphylococci and streptococci intramammary natural infection and inflammation, and subsequently gained a deeper understanding of the immunopathology of mastitis-related infection and inflammation for various etiological agents. ABSTRACT: 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 < 0.05 and FDR < 0.001), and 20 proteins showed significantly discrepant abundance between the genera (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. MDPI 2023-05-31 /pmc/articles/PMC10252062/ /pubmed/37889706 http://dx.doi.org/10.3390/ani13111829 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rešetar Maslov, Dina
Thomas, Funmilola Clara
Beletić, Anđelo
Kuleš, Josipa
Rubić, Ivana
Benić, Miroslav
Bačić, Goran
Maćešić, Nino
Eraghi, Vida
Farkaš, Vladimir
Lenac Roviš, Tihana
Lisnić, Berislav
Žubčić, Damir
Potočnjak, Dalibor
Mrljak, Vladimir
Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics
title Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics
title_full Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics
title_fullStr Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics
title_full_unstemmed Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics
title_short Distinguishing Natural Infections of the Bovine Mammary Gland by Staphylococcus from Streptococcus spp. Using Quantitative Milk Proteomics
title_sort distinguishing natural infections of the bovine mammary gland by staphylococcus from streptococcus spp. using quantitative milk proteomics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10252062/
https://www.ncbi.nlm.nih.gov/pubmed/37889706
http://dx.doi.org/10.3390/ani13111829
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