Circadian Oscillation of Leukocyte Subpopulations and Inflammatory Cytokines over a 24-H Period in Horses
The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood cell count (WBCs), leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes), CD4+, and CD8+ lymphocyte popu...
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Published in | Veterinary sciences Vol. 12; no. 4; p. 386 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
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20.04.2025
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ISSN | 2306-7381 2306-7381 |
DOI | 10.3390/vetsci12040386 |
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Abstract | The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood cell count (WBCs), leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes), CD4+, and CD8+ lymphocyte populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Ten Italian Saddle horses (7–12 years old, body weight 480 ± 30 kg) underwent blood sampling every 4 h over a 24-h period. The COSINOR method was used to identify rhythms and their parameters. A one-way analysis of variance (ANOVA) was applied to identify the differences in acrophase and robustness, and a multiple correlation analysis model (Pearson) was used to evaluate the relationships among the investigated parameters. WBCs, leukocyte subpopulations, CD4+, CD8+, IL-1β, IL-6, and TNFα exhibited daily rhythmicity. In particular, white WBCs, lymphocytes, IL-1β, and IL-6 reached their acrophases during the dark phase, while neuthrophils, CD4+, CD8+, and TNFα showed a diurnal acrophase. One-way ANOVA showed a statistical difference in the acrophase among the investigated parameters (p < 0.0001). The Pearson correlation matrix showed positive and negative relationships among the parameters. Circadian rhythms should be taken into consideration with the daily fluctuations in immune and inflammatory biomarkers to develop good management practices and improve welfare in horses. |
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AbstractList | The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood cell count (WBCs), leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes), CD4+, and CD8+ lymphocyte populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Ten Italian Saddle horses (7–12 years old, body weight 480 ± 30 kg) underwent blood sampling every 4 h over a 24-h period. The COSINOR method was used to identify rhythms and their parameters. A one-way analysis of variance (ANOVA) was applied to identify the differences in acrophase and robustness, and a multiple correlation analysis model (Pearson) was used to evaluate the relationships among the investigated parameters. WBCs, leukocyte subpopulations, CD4+, CD8+, IL-1β, IL-6, and TNFα exhibited daily rhythmicity. In particular, white WBCs, lymphocytes, IL-1β, and IL-6 reached their acrophases during the dark phase, while neuthrophils, CD4+, CD8+, and TNFα showed a diurnal acrophase. One-way ANOVA showed a statistical difference in the acrophase among the investigated parameters (p < 0.0001). The Pearson correlation matrix showed positive and negative relationships among the parameters. Circadian rhythms should be taken into consideration with the daily fluctuations in immune and inflammatory biomarkers to develop good management practices and improve welfare in horses. The temporal organization of biological processes in living organisms follows a specific rhythmicity governed by an endogenous circadian pacemaker, which is an evolutionarily adapted survival mechanism. In recent years, the connection between 24-h biological rhythms, immune pathways, and inflammatory responses has been demonstrated. The present study investigated the daily rhythmicity of white blood cell (WBC) counts, leukocyte subpopulations, CD4+ and CD8+ populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in horses. Statistical analysis showed that WBCs and IL-1β exhibited a nocturnal acrophase, IL-6 and lymphocytes peaked during the early morning hours, and neutrophils, CD4+, CD8+, and TNF-α showed a diurnal acrophase. All parameters showed a robustness of rhythm higher than 70%. This finding is the first step toward understanding how the daily rhythm physiology of horses is closely related to their immunological and inflammatory responses. Studying the rhythms of these parameters can provide a possible window of risk for infections and inflammation to which horses may be susceptible. This is essential for formulating effective management or training regimens for equine athletes, as well as for disease prevention. The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood cell count (WBCs), leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes), CD4+, and CD8+ lymphocyte populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Ten Italian Saddle horses (7–12 years old, body weight 480 ± 30 kg) underwent blood sampling every 4 h over a 24-h period. The COSINOR method was used to identify rhythms and their parameters. A one-way analysis of variance (ANOVA) was applied to identify the differences in acrophase and robustness, and a multiple correlation analysis model (Pearson) was used to evaluate the relationships among the investigated parameters. WBCs, leukocyte subpopulations, CD4+, CD8+, IL-1β, IL-6, and TNFα exhibited daily rhythmicity. In particular, white WBCs, lymphocytes, IL-1β, and IL-6 reached their acrophases during the dark phase, while neuthrophils, CD4+, CD8+, and TNFα showed a diurnal acrophase. One-way ANOVA showed a statistical difference in the acrophase among the investigated parameters ( p < 0.0001). The Pearson correlation matrix showed positive and negative relationships among the parameters. Circadian rhythms should be taken into consideration with the daily fluctuations in immune and inflammatory biomarkers to develop good management practices and improve welfare in horses. The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood cell count (WBCs), leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes), CD4+, and CD8+ lymphocyte populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Ten Italian Saddle horses (7-12 years old, body weight 480 ± 30 kg) underwent blood sampling every 4 h over a 24-h period. The COSINOR method was used to identify rhythms and their parameters. A one-way analysis of variance (ANOVA) was applied to identify the differences in acrophase and robustness, and a multiple correlation analysis model (Pearson) was used to evaluate the relationships among the investigated parameters. WBCs, leukocyte subpopulations, CD4+, CD8+, IL-1β, IL-6, and TNFα exhibited daily rhythmicity. In particular, white WBCs, lymphocytes, IL-1β, and IL-6 reached their acrophases during the dark phase, while neuthrophils, CD4+, CD8+, and TNFα showed a diurnal acrophase. One-way ANOVA showed a statistical difference in the acrophase among the investigated parameters ( < 0.0001). The Pearson correlation matrix showed positive and negative relationships among the parameters. Circadian rhythms should be taken into consideration with the daily fluctuations in immune and inflammatory biomarkers to develop good management practices and improve welfare in horses. The temporal organization of biological processes in living organisms follows a specific rhythmicity governed by an endogenous circadian pacemaker, which is an evolutionarily adapted survival mechanism. In recent years, the connection between 24-h biological rhythms, immune pathways, and inflammatory responses has been demonstrated. The present study investigated the daily rhythmicity of white blood cell (WBC) counts, leukocyte subpopulations, CD4+ and CD8+ populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in horses. Statistical analysis showed that WBCs and IL-1β exhibited a nocturnal acrophase, IL-6 and lymphocytes peaked during the early morning hours, and neutrophils, CD4+, CD8+, and TNF-α showed a diurnal acrophase. All parameters showed a robustness of rhythm higher than 70%. This finding is the first step toward understanding how the daily rhythm physiology of horses is closely related to their immunological and inflammatory responses. Studying the rhythms of these parameters can provide a possible window of risk for infections and inflammation to which horses may be susceptible. This is essential for formulating effective management or training regimens for equine athletes, as well as for disease prevention. The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood cell count (WBCs), leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes), CD4+, and CD8+ lymphocyte populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Ten Italian Saddle horses (7-12 years old, body weight 480 ± 30 kg) underwent blood sampling every 4 h over a 24-h period. The COSINOR method was used to identify rhythms and their parameters. A one-way analysis of variance (ANOVA) was applied to identify the differences in acrophase and robustness, and a multiple correlation analysis model (Pearson) was used to evaluate the relationships among the investigated parameters. WBCs, leukocyte subpopulations, CD4+, CD8+, IL-1β, IL-6, and TNFα exhibited daily rhythmicity. In particular, white WBCs, lymphocytes, IL-1β, and IL-6 reached their acrophases during the dark phase, while neuthrophils, CD4+, CD8+, and TNFα showed a diurnal acrophase. One-way ANOVA showed a statistical difference in the acrophase among the investigated parameters (p < 0.0001). The Pearson correlation matrix showed positive and negative relationships among the parameters. Circadian rhythms should be taken into consideration with the daily fluctuations in immune and inflammatory biomarkers to develop good management practices and improve welfare in horses.The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood cell count (WBCs), leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes), CD4+, and CD8+ lymphocyte populations, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Ten Italian Saddle horses (7-12 years old, body weight 480 ± 30 kg) underwent blood sampling every 4 h over a 24-h period. The COSINOR method was used to identify rhythms and their parameters. A one-way analysis of variance (ANOVA) was applied to identify the differences in acrophase and robustness, and a multiple correlation analysis model (Pearson) was used to evaluate the relationships among the investigated parameters. WBCs, leukocyte subpopulations, CD4+, CD8+, IL-1β, IL-6, and TNFα exhibited daily rhythmicity. In particular, white WBCs, lymphocytes, IL-1β, and IL-6 reached their acrophases during the dark phase, while neuthrophils, CD4+, CD8+, and TNFα showed a diurnal acrophase. One-way ANOVA showed a statistical difference in the acrophase among the investigated parameters (p < 0.0001). The Pearson correlation matrix showed positive and negative relationships among the parameters. Circadian rhythms should be taken into consideration with the daily fluctuations in immune and inflammatory biomarkers to develop good management practices and improve welfare in horses. |
Audience | Academic |
Author | Rizzo, Maria Fazio, Francesco Giannetto, Claudia De Caro, Salvatore Aragona, Francesca Giudice, Elisabetta Arfuso, Francesca Briglia, Marilena Piccione, Giuseppe Costa, Antonino Di Bella, Beatrice |
AuthorAffiliation | 2 Department of Engineering, University of Messina, C/da di Dio (S. Agata), 98166 Messina, Italy; beatrice.dibella@unime.it (B.D.B.); salvatore.decaro@unime.it (S.D.C.) 3 Department of Medicine and Surgery “Kore”, University of Enna, 94100 Enna, Italy; marilena.briglia@unikore.it 1 Department of Veterinary Science, University of Messina, Via Giovanni Palatucci, 98168 Messina, Italy; fraragona@unime.it (F.A.); egiudice@unime.it (E.G.); ffazio@unime.it (F.F.); antonino.costa1@studenti.unime.it (A.C.); farfuso@unime.it (F.A.); gpiccione@unime.it (G.P.); clgiannetto@unime.it (C.G.) |
AuthorAffiliation_xml | – name: 1 Department of Veterinary Science, University of Messina, Via Giovanni Palatucci, 98168 Messina, Italy; fraragona@unime.it (F.A.); egiudice@unime.it (E.G.); ffazio@unime.it (F.F.); antonino.costa1@studenti.unime.it (A.C.); farfuso@unime.it (F.A.); gpiccione@unime.it (G.P.); clgiannetto@unime.it (C.G.) – name: 2 Department of Engineering, University of Messina, C/da di Dio (S. Agata), 98166 Messina, Italy; beatrice.dibella@unime.it (B.D.B.); salvatore.decaro@unime.it (S.D.C.) – name: 3 Department of Medicine and Surgery “Kore”, University of Enna, 94100 Enna, Italy; marilena.briglia@unikore.it |
Author_xml | – sequence: 1 givenname: Francesca orcidid: 0000-0003-2097-0808 surname: Aragona fullname: Aragona, Francesca – sequence: 2 givenname: Maria orcidid: 0000-0003-4343-8937 surname: Rizzo fullname: Rizzo, Maria – sequence: 3 givenname: Elisabetta surname: Giudice fullname: Giudice, Elisabetta – sequence: 4 givenname: Francesco orcidid: 0000-0003-3198-2580 surname: Fazio fullname: Fazio, Francesco – sequence: 5 givenname: Antonino surname: Costa fullname: Costa, Antonino – sequence: 6 givenname: Beatrice orcidid: 0000-0002-1997-4126 surname: Di Bella fullname: Di Bella, Beatrice – sequence: 7 givenname: Salvatore orcidid: 0000-0002-8952-6601 surname: De Caro fullname: De Caro, Salvatore – sequence: 8 givenname: Francesca orcidid: 0000-0001-6139-2691 surname: Arfuso fullname: Arfuso, Francesca – sequence: 9 givenname: Marilena surname: Briglia fullname: Briglia, Marilena – sequence: 10 givenname: Giuseppe orcidid: 0000-0002-4736-1087 surname: Piccione fullname: Piccione, Giuseppe – sequence: 11 givenname: Claudia orcidid: 0000-0003-3049-1181 surname: Giannetto fullname: Giannetto, Claudia |
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Keywords | CD4-CD8 cytokine horse immune system inflammatory response leukocyte subpopulation circadian rhythms interleukins |
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Snippet | The objective of the present study was to investigate the influence of daily rhythms on the immune and inflammatory systems in horses, considering white blood... The temporal organization of biological processes in living organisms follows a specific rhythmicity governed by an endogenous circadian pacemaker, which is an... |
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SubjectTerms | Animal training Antibodies Biological clocks Blood Blood cell count Body weight Catheters CD4 antigen CD4-CD8 CD8 antigen Circadian rhythm Circadian rhythms Cloning Correlation analysis Cytokines Cytotoxicity Diurnal Ethylenediaminetetraacetic acid Flow cytometry horse Horses IL-1β Immune system Immunology Infections Inflammation inflammatory response Interleukin 6 Interleukins leukocyte subpopulation Leukocytes Leukocytes (basophilic) Leukocytes (eosinophilic) Leukocytes (neutrophilic) Lymphocytes Monocytes Pathogens Physiological aspects Physiology Statistical analysis Statistics Tumor necrosis factor Tumor necrosis factor-TNF Tumor necrosis factor-α |
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Title | Circadian Oscillation of Leukocyte Subpopulations and Inflammatory Cytokines over a 24-H Period in Horses |
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