Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean Buffaloes

Brucellosis is a zoonotic disease, with an important economic impact on the livestock industry and public health worldwide. Both Brucella abortus and Brucella melitensis can infect Mediterranean Buffalo (Bubalus bubalis), leading to infertility and abortion. In ruminants, the standard diagnostic app...

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Main Authors: Giulia Franzoni, Federica Signorelli, Anna Donniacuo, Lorena Schiavo, Michele Napoletano, Giovanna De Matteis, Francesco Grandoni, Susanna Zinellu, Vincenzo Bove, Silvia Dei Giudici, Esterina De Carlo, Giorgio Galiero, Francesco Napolitano, Alessandra Martucciello
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Veterinary Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fvets.2025.1583858/full
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author Giulia Franzoni
Federica Signorelli
Anna Donniacuo
Lorena Schiavo
Michele Napoletano
Giovanna De Matteis
Francesco Grandoni
Susanna Zinellu
Vincenzo Bove
Silvia Dei Giudici
Esterina De Carlo
Giorgio Galiero
Francesco Napolitano
Alessandra Martucciello
author_facet Giulia Franzoni
Federica Signorelli
Anna Donniacuo
Lorena Schiavo
Michele Napoletano
Giovanna De Matteis
Francesco Grandoni
Susanna Zinellu
Vincenzo Bove
Silvia Dei Giudici
Esterina De Carlo
Giorgio Galiero
Francesco Napolitano
Alessandra Martucciello
author_sort Giulia Franzoni
collection DOAJ
description Brucellosis is a zoonotic disease, with an important economic impact on the livestock industry and public health worldwide. Both Brucella abortus and Brucella melitensis can infect Mediterranean Buffalo (Bubalus bubalis), leading to infertility and abortion. In ruminants, the standard diagnostic approach involves two serological tests, the Rose Bengal Test and the Complement Fixation Test, applied in parallel, though their specificity requires improvement. Cytokines play a crucial role in coordinating immune responses through complex networks and can serve as biomarkers for various diseases. This study explored the potential use of cytokines as immunological biomarkers for Brucella infection in Mediterranean Buffalo. For this purpose, we included 18 healthy and 20 Brucella-infected buffaloes in our analysis. Heparinized blood samples were stimulated with the Brucella antigen, with PBS as nil control and PWM as lymphocyte viability control. After 16–24 h, plasma levels of IL-1α, IL-1β, IL-4, IL-6, IL-10, IL-17, IL-36Ra, MIP-1α, MIP-1β, MCP-1, CXCL8, IP-10, IFN-γ, TNF, and VEGF-A were measured using multiplex ELISA. Our results showed that infected animals released significantly higher levels of IFN-γ, IP-10, MCP-1 in response to Brucella antigen compared to healthy controls. Conversely, healthy animals released instead higher levels of IL-1α, IL-1β, IL-6 and IL-10 following antigen stimulation compared to infected animals. Finally, sequential canonical discriminant analyses were performed to generate predictive cytokine profiles for each group. The findings indicated that a combination of five cytokines (IFN-γ, IP-10, IL-1α, IL-1β, IL-6) can effectively distinguished infected from healthy buffaloes. Overall, this study suggests that incorporating these key immune cytokines could improve the diagnostic accuracy of brucellosis in Mediterranean Buffalo.
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spelling doaj-art-5042a755dba5470f92104a009b60808d2025-08-20T03:09:24ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692025-05-011210.3389/fvets.2025.15838581583858Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean BuffaloesGiulia Franzoni0Federica Signorelli1Anna Donniacuo2Lorena Schiavo3Michele Napoletano4Giovanna De Matteis5Francesco Grandoni6Susanna Zinellu7Vincenzo Bove8Silvia Dei Giudici9Esterina De Carlo10Giorgio Galiero11Francesco Napolitano12Alessandra Martucciello13Department of Animal Health, Istituto Zooprofilattico Sperimentale della Sardegna, Sassari, ItalyCouncil for Agricultural Research and Economics (CREA)- Research Centre for Animal Production and Aquaculture, Monterotondo (RM), ItalyNational Reference Centre for Hygiene and Technologies of Mediterranean Buffalo Farming and Productions, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Salerno, ItalyNational Reference Centre for Hygiene and Technologies of Mediterranean Buffalo Farming and Productions, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Salerno, ItalyNational Reference Centre for Hygiene and Technologies of Mediterranean Buffalo Farming and Productions, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Salerno, ItalyCouncil for Agricultural Research and Economics (CREA)- Research Centre for Animal Production and Aquaculture, Monterotondo (RM), ItalyCouncil for Agricultural Research and Economics (CREA)- Research Centre for Animal Production and Aquaculture, Monterotondo (RM), ItalyDepartment of Animal Health, Istituto Zooprofilattico Sperimentale della Sardegna, Sassari, ItalyNational Reference Centre for Hygiene and Technologies of Mediterranean Buffalo Farming and Productions, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Salerno, ItalyDepartment of Animal Health, Istituto Zooprofilattico Sperimentale della Sardegna, Sassari, ItalyNational Reference Centre for Hygiene and Technologies of Mediterranean Buffalo Farming and Productions, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Salerno, ItalyNational Reference Centre for Hygiene and Technologies of Mediterranean Buffalo Farming and Productions, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Salerno, ItalyCouncil for Agricultural Research and Economics (CREA)- Research Centre for Animal Production and Aquaculture, Monterotondo (RM), ItalyNational Reference Centre for Hygiene and Technologies of Mediterranean Buffalo Farming and Productions, Istituto Zooprofilattico Sperimentale del Mezzogiorno, Salerno, ItalyBrucellosis is a zoonotic disease, with an important economic impact on the livestock industry and public health worldwide. Both Brucella abortus and Brucella melitensis can infect Mediterranean Buffalo (Bubalus bubalis), leading to infertility and abortion. In ruminants, the standard diagnostic approach involves two serological tests, the Rose Bengal Test and the Complement Fixation Test, applied in parallel, though their specificity requires improvement. Cytokines play a crucial role in coordinating immune responses through complex networks and can serve as biomarkers for various diseases. This study explored the potential use of cytokines as immunological biomarkers for Brucella infection in Mediterranean Buffalo. For this purpose, we included 18 healthy and 20 Brucella-infected buffaloes in our analysis. Heparinized blood samples were stimulated with the Brucella antigen, with PBS as nil control and PWM as lymphocyte viability control. After 16–24 h, plasma levels of IL-1α, IL-1β, IL-4, IL-6, IL-10, IL-17, IL-36Ra, MIP-1α, MIP-1β, MCP-1, CXCL8, IP-10, IFN-γ, TNF, and VEGF-A were measured using multiplex ELISA. Our results showed that infected animals released significantly higher levels of IFN-γ, IP-10, MCP-1 in response to Brucella antigen compared to healthy controls. Conversely, healthy animals released instead higher levels of IL-1α, IL-1β, IL-6 and IL-10 following antigen stimulation compared to infected animals. Finally, sequential canonical discriminant analyses were performed to generate predictive cytokine profiles for each group. The findings indicated that a combination of five cytokines (IFN-γ, IP-10, IL-1α, IL-1β, IL-6) can effectively distinguished infected from healthy buffaloes. Overall, this study suggests that incorporating these key immune cytokines could improve the diagnostic accuracy of brucellosis in Mediterranean Buffalo.https://www.frontiersin.org/articles/10.3389/fvets.2025.1583858/fullBrucellabiomarkersIFN-γcytokinesMediterranean Buffaloes
spellingShingle Giulia Franzoni
Federica Signorelli
Anna Donniacuo
Lorena Schiavo
Michele Napoletano
Giovanna De Matteis
Francesco Grandoni
Susanna Zinellu
Vincenzo Bove
Silvia Dei Giudici
Esterina De Carlo
Giorgio Galiero
Francesco Napolitano
Alessandra Martucciello
Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean Buffaloes
Frontiers in Veterinary Science
Brucella
biomarkers
IFN-γ
cytokines
Mediterranean Buffaloes
title Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean Buffaloes
title_full Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean Buffaloes
title_fullStr Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean Buffaloes
title_full_unstemmed Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean Buffaloes
title_short Exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in Mediterranean Buffaloes
title_sort exploring potential cytokine profiles as diagnostic biomarkers for brucellosis in mediterranean buffaloes
topic Brucella
biomarkers
IFN-γ
cytokines
Mediterranean Buffaloes
url https://www.frontiersin.org/articles/10.3389/fvets.2025.1583858/full
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