Publication: Análisis del estado de salud porcino en condiciones de campo mediante el uso de biomarcadores salivales
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Date
2025-12-01
Authors
Sánchez Hernández, Jorge
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Escuela Internacional de Doctorado
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Gutiérrez Montes, Ana María
Publisher
Universidad de Murcia
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DOI
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info:eu-repo/semantics/doctoralThesis
Description
Abstract
Esta Tesis Doctoral aporta una visión global en el análisis del estado de salud del ganado porcino en condiciones de campo mediante el uso de biomarcadores salivales.
Mediante cuatro experimentales se buscaron tres objetivos: (i) aumentar el conocimiento de los biomarcadores salivales actuales, establecer sus intervalos de referencia y valorar nuevos biomarcadores (ii) comparar el poder diagnóstico de la saliva frente al suero para detectar enfermedad con la ventaja de usar una muestra no invasiva (iii) diferenciar distintas alteraciones de la homeostasis porcina mediante un panel de biomarcadores salivales.
El objetivo 1 se explica en las experimentales 1 y 2 mediante el análisis de los factores que influencian los valores normales de animales sanos y la cuantificación, por primera vez, de elementos traza en muestras de saliva porcina como biomarcadores de salud. En la experimental 1 se evaluó la influencia de la raza, sexo y etapa de producción en los valores normales de los biomarcadores salivales proteína C reactiva (CRP), haptoglobina (Hp), adenosina desaminasa (ADA) y capacidad antioxidante total (TAC). Se utilizaron cerdos comerciales e ibéricos observándose diferencias en todos los marcadores estudiados. Los factores sexo y etapa de producción se analizaron por separado para cada raza, encontrándose también diferencias. Se podría concluir que la raza, el sexo y la etapa de producción influyen en los valores de los biomarcadores salivales del estado de salud en cerdos y deben tenerse en cuenta para establecer intervalos de referencia y poder realizar interpretaciones correctas en condiciones de campo.
La experimental 2 cuantificó los elementos traza Cu, Zn, Se, Cd y Pb en saliva porcina mediante espectrometría de absorción atómica en animales con diversas patologías (cojeras, fatigas, prolapsos rectales o retraso en el crecimiento). Se estudió la posible asociación entre los valores de elementos traza y los biomarcadores salivales de estado de salud CRP, Hp, TAC y ADA encontrándose alteraciones en los niveles de dos o más elementos traza en los distintos grupos de cerdos enfermos, con variaciones según patología, por lo que la cuantificación salival de oligoelementos podría considerarse una herramienta complementaria para la detección y diferenciación de enfermedades.
El segundo objetivo se explicó mediante la experimental 3, donde se comparó el poder diagnóstico de un conjunto de biomarcadores (CRP, Hp, ADA, TAC, Cu, Zn y proteínas totales) en muestras de saliva y suero. Los resultados mostraron que la saliva podría reemplazar al suero para la medición de proteínas de fase aguda. Para otros biomarcadores no hubo correlación, pero el análisis en saliva mostró una sensibilidad superior para detectar enfermedades que el suero. Además, la mejor combinación de biomarcadores en saliva (Hp, CRP, TAC y ADA) requiere menor tiempo, dinero y cantidad de muestra que la mejor combinación en suero (Hp, CRP y Cu). Asimismo, la distribución de biomarcadores analizados mostró una mayor diferencia entre animales sanos y enfermos cuando se utilizaron muestras de saliva. En resumen, la saliva podría considerarse una alternativa eficiente al suero para la detección de cerdos enfermos.
El objetivo 3 se alcanza mediante la experimental 4, donde se analizó la asociación entre los biomarcadores salivales de estrés psicosocial (cortisol, alfa-amilasa) y oxidativo [(TAC, estado oxidante total (TOS) e índice de estrés oxidativo (OSI)] con la respuesta inmune innata (CRP, Hp) y adaptativa (ADA, Cu, Zn), realizando un primer intento de discriminar entre enfermedades (mordeduras de colas, prolapsos, diarreas, cojeras y disneas).
Se encontró una asociación alta entre estrés oxidativo y sistema inmune adaptativo y moderada entre estrés psicosocial y sistema inmune. Cada condición patológica mostró diferencias en el grado de activación de los sistemas analizados, lo que podría utilizarse para crear diferentes perfiles y caracterizar enfermedades.
This Doctoral Thesis provides a comprehensive view of the health status of swine under field conditions through the use of salivary biomarkers. Through four experimental studies, three objectives were pursued: (i) to increase knowledge of current salivary biomarkers, establish their reference ranges, and evaluate new biomarkers, (ii) to compare the diagnostic power of saliva versus serum for disease detection, leveraging the advantage of a non-invasive sample, and (iii) to attempt to differentiate various alterations in swine homeostasis using a panel of salivary biomarkers. Objective 1 is addressed in experiments 1 and 2 by analyzing the factors that influence normal values in healthy animals and, for the first time, quantifying trace elements in porcine saliva samples as health biomarkers. In experiment 1, the influence of breed, sex, and production stage on normal values of salivary biomarkers, such as C-reactive protein (CRP), haptoglobin (Hp), adenosine deaminase (ADA), and total antioxidant capacity (TAC), was evaluated. Commercial and Iberian pigs were used, revealing differences in all studied biomarkers. Sex and production stage were analyzed separately for each breed, also revealing differences. It can be concluded that breed, sex, and production stage influence salivary biomarker values for health status in pigs and should be considered when establishing reference ranges for accurate field interpretations. Experiment 2 quantified trace elements such as Cu, Zn, Se, Cd, and Pb in porcine saliva using atomic absorption spectrometry in animals with various pathologies (lameness, fatigue, rectal prolapse, or growth retardation). The possible association between trace element values and salivary health biomarkers CRP, Hp, TAC and ADA was studied. Alterations in the levels of two or more trace elements were found in the different groups of sick pigs, with variations depending on the pathology. Therefore, salivary quantification of trace elements could be considered a complementary tool for disease detection and differentiation. The second objective was addressed in experiment 3, where the diagnostic power of a set of biomarkers (CRP, Hp, ADA, TAC, Cu, Zn and total protein (PT)) in saliva and serum samples was compared. Results showed that saliva could replace serum for measuring acute-phase proteins. For other biomarkers, there was no correlation, but saliva analysis showed superior sensitivity for disease detection compared to serum. Moreover, the best combination of salivary biomarkers (Hp, CRP, TAC and ADA) required less time, money and sample volume than the best serum combination (Hp, CRP and Cu). Additionally, the distribution of all analyzed biomarkers showed greater differences between healthy and diseased animals when saliva samples were used. In summary, saliva could be considered an efficient alternative to serum for detecting diseased pigs. Objective 3 was achieved through experiment 4, where the association between salivary biomarkers of psychosocial stress (cortisol, alpha-amylase) and oxidative stress [(TAC, total oxidant status (TOS), and oxidative stress index (OSI)] with innate immune response (CRP, Hp) and adaptive immune response (ADA, Cu, Zn, PT) was analyzed, attempting to discriminate between diseases (tail biting, prolapse, diarrhea, lameness, and dyspnea). A strong association was found between oxidative stress and the adaptive immune system, and a moderate association between psychosocial stress and the immune system. Each pathological condition showed differences in the degree of system activation, which could be used to create different profiles and characterize diseases.
This Doctoral Thesis provides a comprehensive view of the health status of swine under field conditions through the use of salivary biomarkers. Through four experimental studies, three objectives were pursued: (i) to increase knowledge of current salivary biomarkers, establish their reference ranges, and evaluate new biomarkers, (ii) to compare the diagnostic power of saliva versus serum for disease detection, leveraging the advantage of a non-invasive sample, and (iii) to attempt to differentiate various alterations in swine homeostasis using a panel of salivary biomarkers. Objective 1 is addressed in experiments 1 and 2 by analyzing the factors that influence normal values in healthy animals and, for the first time, quantifying trace elements in porcine saliva samples as health biomarkers. In experiment 1, the influence of breed, sex, and production stage on normal values of salivary biomarkers, such as C-reactive protein (CRP), haptoglobin (Hp), adenosine deaminase (ADA), and total antioxidant capacity (TAC), was evaluated. Commercial and Iberian pigs were used, revealing differences in all studied biomarkers. Sex and production stage were analyzed separately for each breed, also revealing differences. It can be concluded that breed, sex, and production stage influence salivary biomarker values for health status in pigs and should be considered when establishing reference ranges for accurate field interpretations. Experiment 2 quantified trace elements such as Cu, Zn, Se, Cd, and Pb in porcine saliva using atomic absorption spectrometry in animals with various pathologies (lameness, fatigue, rectal prolapse, or growth retardation). The possible association between trace element values and salivary health biomarkers CRP, Hp, TAC and ADA was studied. Alterations in the levels of two or more trace elements were found in the different groups of sick pigs, with variations depending on the pathology. Therefore, salivary quantification of trace elements could be considered a complementary tool for disease detection and differentiation. The second objective was addressed in experiment 3, where the diagnostic power of a set of biomarkers (CRP, Hp, ADA, TAC, Cu, Zn and total protein (PT)) in saliva and serum samples was compared. Results showed that saliva could replace serum for measuring acute-phase proteins. For other biomarkers, there was no correlation, but saliva analysis showed superior sensitivity for disease detection compared to serum. Moreover, the best combination of salivary biomarkers (Hp, CRP, TAC and ADA) required less time, money and sample volume than the best serum combination (Hp, CRP and Cu). Additionally, the distribution of all analyzed biomarkers showed greater differences between healthy and diseased animals when saliva samples were used. In summary, saliva could be considered an efficient alternative to serum for detecting diseased pigs. Objective 3 was achieved through experiment 4, where the association between salivary biomarkers of psychosocial stress (cortisol, alpha-amylase) and oxidative stress [(TAC, total oxidant status (TOS), and oxidative stress index (OSI)] with innate immune response (CRP, Hp) and adaptive immune response (ADA, Cu, Zn, PT) was analyzed, attempting to discriminate between diseases (tail biting, prolapse, diarrhea, lameness, and dyspnea). A strong association was found between oxidative stress and the adaptive immune system, and a moderate association between psychosocial stress and the immune system. Each pathological condition showed differences in the degree of system activation, which could be used to create different profiles and characterize diseases.
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