Blood proteome profiling for biomarker discovery in broilers with necrotic enteritis

Abstract Analysis of the blood proteome allows identification of proteins related to changes upon certain physiological conditions. The pathophysiology of necrotic enteritis (NE) has been extensively studied. While intestinal changes have been very well documented, data addressing NE-induced alterat...

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Main Authors: Svitlana Tretiak, Teresa Mendes Maia, Delphi Van Haver, An Staes, Simon Devos, Tom Rijsselaere, Evy Goossens, Filip Van Immerseel, Francis Impens, Gunther Antonissen
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-97783-w
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author Svitlana Tretiak
Teresa Mendes Maia
Delphi Van Haver
An Staes
Simon Devos
Tom Rijsselaere
Evy Goossens
Filip Van Immerseel
Francis Impens
Gunther Antonissen
author_facet Svitlana Tretiak
Teresa Mendes Maia
Delphi Van Haver
An Staes
Simon Devos
Tom Rijsselaere
Evy Goossens
Filip Van Immerseel
Francis Impens
Gunther Antonissen
author_sort Svitlana Tretiak
collection DOAJ
description Abstract Analysis of the blood proteome allows identification of proteins related to changes upon certain physiological conditions. The pathophysiology of necrotic enteritis (NE) has been extensively studied. While intestinal changes have been very well documented, data addressing NE-induced alterations in the blood proteome are scant, although these might have merit in diagnostics. In light of recent technological advancements in proteomics and pressing need for tools to access gut health, the current study employs mass-spectrometry (MS)-based proteomics to identify biomarkers for gastrointestinal health of chickens. Here, we report findings of an untargeted proteomics investigation conducted on blood plasma in chickens under NE challenge. Two MS-strategies were used for analysis: conventional data dependent acquisition coupled to standard nanoflow liquid chromatography (LC) (nano-DDA) and recently-developed data independent acquisition coupled to an Evosep One LC system (Evo-DIA). Despite superior completeness and quantification of the Evo-DIA-acquired data, high degree of agreement in identification and quantification was observed between both approaches. Additionally, we identified 15 differentially expressed proteins (shared by nano-DDA and Evo-DIA) that represent responses of animals to infection and may serve as potential biomarkers. Experimental validation through ELISA immunoassays and targeted MS for selected regulated proteins (CFD, HPS5, and MASP2) confirmed medium-to-high levels of inter-protein correlation. A GSEA analysis revealed enrichment in a number of processes related to adaptive and humoral immunity, immune activation and response in infected animals. Data are available via ProteomeXchange with identifiers PXD050461, PXD050473, and PXD061607.
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spelling doaj-art-ec9eb980da7c4584936cd9e917a3d1d32025-08-20T02:27:53ZengNature PortfolioScientific Reports2045-23222025-04-0115111710.1038/s41598-025-97783-wBlood proteome profiling for biomarker discovery in broilers with necrotic enteritisSvitlana Tretiak0Teresa Mendes Maia1Delphi Van Haver2An Staes3Simon Devos4Tom Rijsselaere5Evy Goossens6Filip Van Immerseel7Francis Impens8Gunther Antonissen9Livestock Gut Health Team (LiGHT) Ghent, Department of Pathobiology, Pharmacology and Zoological Medicine, Faculty of Veterinary Medicine, Ghent UniversityVIB-UGent Center for Medical Biotechnology, VIBVIB-UGent Center for Medical Biotechnology, VIBVIB-UGent Center for Medical Biotechnology, VIBVIB-UGent Center for Medical Biotechnology, VIBImpextraco NVLivestock Gut Health Team (LiGHT) Ghent, Department of Pathobiology, Pharmacology and Zoological Medicine, Faculty of Veterinary Medicine, Ghent UniversityLivestock Gut Health Team (LiGHT) Ghent, Department of Pathobiology, Pharmacology and Zoological Medicine, Faculty of Veterinary Medicine, Ghent UniversityVIB-UGent Center for Medical Biotechnology, VIBLivestock Gut Health Team (LiGHT) Ghent, Department of Pathobiology, Pharmacology and Zoological Medicine, Faculty of Veterinary Medicine, Ghent UniversityAbstract Analysis of the blood proteome allows identification of proteins related to changes upon certain physiological conditions. The pathophysiology of necrotic enteritis (NE) has been extensively studied. While intestinal changes have been very well documented, data addressing NE-induced alterations in the blood proteome are scant, although these might have merit in diagnostics. In light of recent technological advancements in proteomics and pressing need for tools to access gut health, the current study employs mass-spectrometry (MS)-based proteomics to identify biomarkers for gastrointestinal health of chickens. Here, we report findings of an untargeted proteomics investigation conducted on blood plasma in chickens under NE challenge. Two MS-strategies were used for analysis: conventional data dependent acquisition coupled to standard nanoflow liquid chromatography (LC) (nano-DDA) and recently-developed data independent acquisition coupled to an Evosep One LC system (Evo-DIA). Despite superior completeness and quantification of the Evo-DIA-acquired data, high degree of agreement in identification and quantification was observed between both approaches. Additionally, we identified 15 differentially expressed proteins (shared by nano-DDA and Evo-DIA) that represent responses of animals to infection and may serve as potential biomarkers. Experimental validation through ELISA immunoassays and targeted MS for selected regulated proteins (CFD, HPS5, and MASP2) confirmed medium-to-high levels of inter-protein correlation. A GSEA analysis revealed enrichment in a number of processes related to adaptive and humoral immunity, immune activation and response in infected animals. Data are available via ProteomeXchange with identifiers PXD050461, PXD050473, and PXD061607.https://doi.org/10.1038/s41598-025-97783-wBroiler chickenDDADIAProteomicsNecrotic enteritisGastrointestinal health
spellingShingle Svitlana Tretiak
Teresa Mendes Maia
Delphi Van Haver
An Staes
Simon Devos
Tom Rijsselaere
Evy Goossens
Filip Van Immerseel
Francis Impens
Gunther Antonissen
Blood proteome profiling for biomarker discovery in broilers with necrotic enteritis
Scientific Reports
Broiler chicken
DDA
DIA
Proteomics
Necrotic enteritis
Gastrointestinal health
title Blood proteome profiling for biomarker discovery in broilers with necrotic enteritis
title_full Blood proteome profiling for biomarker discovery in broilers with necrotic enteritis
title_fullStr Blood proteome profiling for biomarker discovery in broilers with necrotic enteritis
title_full_unstemmed Blood proteome profiling for biomarker discovery in broilers with necrotic enteritis
title_short Blood proteome profiling for biomarker discovery in broilers with necrotic enteritis
title_sort blood proteome profiling for biomarker discovery in broilers with necrotic enteritis
topic Broiler chicken
DDA
DIA
Proteomics
Necrotic enteritis
Gastrointestinal health
url https://doi.org/10.1038/s41598-025-97783-w
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