María Botía

In this report, a comprehensive panel of analytes including biomarkers of stress, reproduction related hormones, biomarkers of inflammation and immunity, oxidative stress biomarkers, minerals and enzymes were monitored daily from 3 days before farrowing until the day of farrowing in the saliva of 23 healthy sows. All the analytes with exception of testosterone, serum amyloid-A, uric acid, calcium and phosphorous showed an increase the day of farrowing. Cortisol, cortisone, oxytocin, estradiol, haptoglobin, the cupric reducing antioxidant capacity and zinc showed increases the day just before farrowing compared to the previous days. In the conditions of our report, a single cortisol value higher than 78.5 ng/dL would indicate that farrowing could occur in the following 24 hours with 89% accuracy. When the magnitude of change observed between two consecutive days were evaluated, cortisol showed 82.6% accuracy for detecting farrowing in the following 24 hours when more than 1.57-fold increase was observed between the two days. The results indicated that there are analytes in saliva that could change in the previous days to farrowing and that an increase in cortisol could potentially be used as a biomarkers of the delivery. These results should be confirmed with a larger population.

In this report, a comprehensive panel of analytes including biomarkers of stress, reproduction related hormones, biomarkers of inflammation and immunity, oxidative stress biomarkers, minerals and enzymes were monitored daily from 3 days before farrowing until the day of farrowing in the saliva of 23 healthy sows. All the analytes with exception of testosterone, serum amyloid-A, uric acid, calcium and phosphorous showed an increase the day of farrowing. Cortisol, cortisone, oxytocin, estradiol, haptoglobin, the cupric reducing antioxidant capacity and zinc showed increases the day just before farrowing compared to the previous days. In the conditions of our report, a single cortisol value higher than 78.5 ng/dL would indicate that farrowing could occur in the following 24 hours with 89% accuracy. When the magnitude of change observed between two consecutive days were evaluated, cortisol showed 82.6% accuracy for detecting farrowing in the following 24 hours when more than 1.57-fold increase was observed between the two days. The results indicated that there are analytes in saliva that could change in the previous days to farrowing and that an increase in cortisol could potentially be used as a biomarkers of the delivery. These results should be confirmed with a larger population.

Background: Oxidative stress contributes to the development and progression of epilepsy and is connected with neuroinflammation during epileptic seizures. Cholinesterase has a modulatory role, and oxytocin has antiepileptic properties. The purpose of this study was to assess selective inflammatory (C-Reactive Protein, CRP) and oxidative stress markers [Paraoxonase-1 (PON1), cupric reducing antioxidant capacity (CUPRAC), ferric reducing antioxidant power (FRAP), cholinesterase, and oxytocin in serum and cerebrospinal fluid (CSF) samples of dogs with different types of epilepsy. Methods: There were four groups of dogs; A: healthy controls; B: idiopathic epilepsy receiving antiepileptic medication; C: idiopathic epilepsy without antiepileptic medication; and D: structural epilepsy. CRP, PON1, CUPRAC, and cholinesterase were evaluated in serum and PON1, CUPRAC, FRAP, cholinesterase and oxytocin were evaluated in CSF samples. Group differences were evaluated using the ANOVA test, followed by post hoc analyses or Kruskal–Wallis/Dunn’s test. Results: Fifty-one serum and 26 CSF samples were analyzed. CSF PON1 was significantly different in group D compared with groups A and C (p = 0.044 and p = 0.008, respectively). CSF cholinesterase was significantly different in group D compared with groups A, B and C (p = 0.003, p = 0.025, and p = 0.033, respectively). Conclusions: Structural epilepsy may influence PON1 and cholinesterase levels in CSF samples. Compared with CSF, serum was not the most suitable biological material to investigate oxidative stress and inflammatory markers.

The intensification of pig ( Sus scrofa domesticus ) production systems raises concerns regarding animal welfare, particularly during pre-slaughter conditions, a phase associated with significant stress. Saliva is increasingly recognised as a non-invasive matrix for detecting stress-related biomarkers in pigs. This preliminary study aimed to explore salivary protein changes in pigs subjected to two distinct pre-slaughter conditions at the slaughterhouse, improved (Group A) and stressful (Group B), by tandem mass tag (TMT)-based proteomics. Proteomic analysis of saliva from three pigs per group revealed 13 proteins with a statistically significant difference in relative abundance between the groups. Group B showed elevated levels of proteins linked to metabolic stress, inflammation, and coagulation, such as cystatin-C and fibrinogen chains, while proteins like vimentin and follistatin-related protein were decreased. Cystatin-C and vimentin were further validated by immunoassays in 12 additional pigs per group, confirming their differential abundance. These findings suggest that salivary cystatin-C and vimentin, along with the other 11 proteins that showed changes at proteomics, may serve as candidate biomarkers of acute stress at slaughter. While further validation is required, our results support the potential of salivary proteomics for welfare monitoring in livestock.

Weaning is one of the most stressful stages in pig production, especially for Iberian piglets, which grow more slowly than other cosmopolitan breeds and therefore, have a lower weaning weight when raised in intensive systems. Stress at weaning, caused by separation from the sow, dietary change, and regrouping with unfamiliar piglets, can negatively impact welfare, immune function, and performance. Pre-weaning socialization, which allows piglets from different litters to interact before weaning, has been proposed as a strategy to reduce aggression and facilitate the adaptation to the post-weaning period. However, its physiological effects in Iberian pigs remain largely unknown. In this study, 8 Iberian sows and their litters were assigned to either a control group (CTRL) or a socialization group (SOC) where litters were mingled (socialized) two weeks before weaning. Salivary and serum biomarkers of stress, inflammation, immunity and metabolism were measured at weaning (pwD0) and 7 days post-weaning (pwD7), and growth performance was recorded until 60 days of age. Socialized piglets showed reduced salivary Adenosine Deaminase (ADA) activity at pwD0 and pwD7 and lower salivary chromogranin A (CgA) and serum Haptoglobin (Hp) levels at pwD7. In contrast, they presented higher concentrations in serum of high-density lipoprotein (HDL), low-density lipoprotein (LDL), total cholesterol, glucose, and urea at pwD7 compared to controls. Attending to the sex effect, Butyryl-cholinesterase (BChE) serum concentration was higher in males and urea, and creatinine were higher in females. Growth rates were higher in socialized piglets in the first two weeks post-weaning but lower thereafter. These findings may suggest that pre-weaning socialization could reduce the stress associated with early post-weaning in Iberian piglets, thus potentially improving welfare and adaptation during this period.

Background: Oxidative stress contributes to the development and progression of epilepsy and is connected with neuroinflammation during epileptic seizures. Cholinesterase has a modulatory role, and oxytocin has antiepileptic properties. The purpose of this study was to assess selective inflammatory (CRP) and oxidative stress markers (PON1, CUPRAC, FRAP), cholinesterase, and oxytocin in serum and CSF samples of dogs with different types of epilepsy. Methods: There were four groups of dogs; A: healthy controls; B: idiopathic epilepsy receiving antiepileptic medication; C: idiopathic epilepsy without antiepileptic medication; and D: structural epilepsy. CRP, PON1, CUPRAC, and cholinesterase were evaluated in serum and PON1, CUPRAC, FRAP, cholinesterase and oxytocin were evaluated in CSF samples. Group differences were evaluated using ANOVA or Kruskal–Wallis tests, followed by post-hoc analyses. Results: Fifty-one serum and 26 CSF samples were analyzed. CSF PON1 was significantly different in group D compared to groups A and C (p=0.044 and p=0.008, respectively). CSF cholinesterase was significantly different in group D compared to groups A, B and C (p=0.003, p=0.025, p=0.033, respectively). Conclusions: Structural epilepsy may influence PON1, CUPRAC and cholinesterase levels in CSF samples. Compared to CSF, serum was not the most suitable biological material to investigate oxidative stress and inflammatory markers.

Equine Gastric Ulcer Syndrome (EGUS) is a widespread disease with a very high prevalence and importance in horses. It includes two conditions: Equine squamous gastric disease (ESGD), linked to acid disturbances, and Equine Glandular Gastric Disease (EGGD), associated with inflammation and immune system issues. In saliva, increased levels of immunological markers have been observed in this disease, indicating immune system involvement. In addition, changes in biomarkers of oxidative stress have been described. In horses, IgG and IgA are proteins that appear in saliva and serum and are considered to be the major proteins in specific immunity; however, their levels in horses with EGUS have not been previously studied. This report aims to evaluate IgG and IgA concentrations in the saliva and serum of horses with EGUS, measured by automated assays, and compare them with other biomarkers of the immune and oxidative stress system. This work validated the IgG and IgA assays in saliva and showed that IgA significantly increased in the saliva of horses with EGUS. When the correlation between IgG and IgA and other biomarkers of immune system issues and oxidative stress such as ADA, S100 A8-A9 (calprotectin), S100 A12 (calgranulin), uric acid, FRAS, and AOPP was studied, IgA in saliva showed a significant moderate correlation with adenosine deaminase, indicating its possible involvement in the immune reaction occurring in EGUS.

Colostrum is a mammary secretion released from the time of farrowing to 36 h post-farrowing. After this time and during all the rest of lactation, the mammary secretion is considered milk. The objectives of this study were: (1) to perform an analytical validation in the colostrum and milk of sows of assays for four analytes related to immunity: total ADA (tADA) and its isoenzymes (ADA1 and ADA2), myeloperoxidase (Mpx), calprotectin, and calgranulin, and two analytes related to welfare: cortisol and alpha-amylase. (2) To evaluate the changes in these analytes during lactation (3) To assess the correlations between these new analytes, as well as with IgG and IgA. In the analytical validation, all the assays were precise and accurate. When changes during lactation were evaluated, the concentration of tADA and ADA2 was found to be higher in colostrum than in milk (p < 0.02), while the activity of Mpx was observed to be higher in mature milk than in colostrum (p < 0.03). Furthermore, cortisol and alpha-amylase activity were found to be higher in colostrum compared to mature milk (p < 0.04 and p < 0.0001, respectively). Regarding the relation between analytes, alpha-amylase showed a significant correlation with both IgG and IgA and calprotectin was correlated with calgranulin and Mpx. Further studies should be performed to elucidate the possible practical application of the analytes evaluated in this study as biomarkers of colostrum and milk in sows.