Trace Elements and Ferritin in Pig Saliva : Variations during Fattening, Time of Sampling, Effect of Dirtiness and Stability under Different Storage Conditions

Altres ajuts: The European Next Generation Funds RYC2021-033660-IThe objective of this study was to evaluate the possible changes of zinc (Zn), copper (Cu), iron (Fe) and ferritin during the entire productive cycle in fattening pigs and at different diurnal sampling times. Moreover, the possible effects of the presence of pen contaminants and storage stability at different temperature conditions were assessed. The analytes changed along the different phases of the fattening productive cycle, showing, in general, higher values at the initial phases. In addition, statistically significant variations were found in Zn and Cu measurements at different sampling times of the day. In the spectrophotometric assays, the values of all analytes significantly increased after adding high concentrations of feces or feed. However, when low concentrations of feces or feed were added, only Cu showed a significant increase. Overall, the salivary levels of Zn, Cu, Fe and ferritin in pigs can change during different fattening phases and the different hours of the day. These analytes were more stable at −80 °C and, if saliva is contaminated with feces or feed, it can lead to an increase in these analytes

Prolactin in saliva of pigs as a possible biomarker of stress: analytical validation of an immunoassay and changes at slaughterhouse

Oral session 2[EN] Prolactin (PRL) is considered a biomarker of stress response, both acute and chronic, in several species. Although the scientific literature reports divergent results, some studies suggest that prolactin secretion during stress acts to maintain homeostasis within the immune system. Several studies have investigated the possibility of measuring PRL in saliva in different species such as rats, domestic ruminants, donkeys, dogs, cattle, and sheep for diagnosis or research purposes. Results obtained in animal models have indicated that the secretion of this hormone is dependent on the type and intensity of the stress

Investigation of correlations between skin lesion count and concentrations of salivary biomarkers in pigs from suckling to fattening

Oral session 1[EN] ClearFarm project intends to develop a platform to monitor animal welfare continuously throughout the value chain of pigs and dairy cattle using sensor technology. This platform relies on algorithms (for each species), built up with relevant welfare indicators measured by PLF (Precision Livestock Farming) technology

Evaluation of the Effect of Sampling Time on Biomarkers of Stress, Immune System, Redox Status and Other Biochemistry Analytes in Saliva of Finishing Pigs

Saliva is a sample with a high potential in pigs since it is usually easy to obtain and its collection from animals causes less stress than blood sampling. However, the possible effects of daily variations in many salivary biomarkers are still unknown in this species. In our report, the possible variations depending on the sampling time in the day in a panel of 26 salivary biomarkers related to stress, immune system, redox status and other physiological functions in the saliva of pigs were evaluated. In our experimental conditions, daily variations were observed in cortisol, α-amylase, total esterase, butyrylcholinesterase, lipase, adenosine deaminase isoenzyme 1, uric acid, superoxide dismutase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatine kinase, lactate dehydrogenase, triglycerides and lactate. In some analytes, these differences appeared in both sexes, whereas others only showed differences in one sex. These variations should be taken into consideration for an appropriate interpretation of these analytes in the saliva of healthy pigs. This study aims to evaluate the possible variations due to the sampling time in the day in 26 analytes of pigs' saliva, related to stress, the immune system, redox status and other biomarkers related to metabolism and selected tissues and organs, in order to know the possible effects of the hour of the day in their interpretation. These analytes were measured in saliva obtained from a population of 40 clinically healthy pigs from 8 a.m. to 8 p.m., every 4 h in the same day. In our experimental conditions, daily variations were observed in cortisol, salivary α-amylase, total esterase activity, butyrylcholinesterase, lipase, adenosine deaminase isoenzyme 1, uric acid, superoxide dismutase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatine kinase, lactate dehydrogenase, lactate and triglycerides. These changes appeared in both sexes, except for adenosine deaminase isoenzyme 1 and superoxide dismutase which only showed differences in females. In conclusion, this report indicates that, in the experimental conditions of this trial, the time of the day and sex can influence the values obtained in various salivary analytes in pigs. These variations should thus be taken into consideration for an adequate interpretation of these analytes when used for the evaluation of health and welfare in this species

Analysing biomarkers in oral fluid from pigs: influence of collection strategy and age of the pig

peer-reviewedBackground and objectives Oral fluid (OF) is an easy-to-collect, inexpensive, fast and non-invasive sample to characterize health and welfare status of the pig. However, further standardisation of the collection methods is needed in order to use it regularly in veterinary practice. Cotton ropes are routinely used to collect OF for pathogen detection but they may not be optimal for biomarker analysis due to sample contamination. This study compared two methods (cotton ropes and sponges) to collect porcine OF for biomarker analysis. A panel of 11 biomarkers of stress, inflammation, sepsis, immunity, redox status and general homeostasis was studied. Materials and methods Eighteen farrow-to-finish pig farms were included in the study. In each farm, three (for sponges) or four pens of pigs (for ropes) were sampled at four age categories: the week after weaning (5 weeks), before (11–12 weeks) and after (12–13 weeks) moving to finisher facility and the week before slaughter (22–25 weeks). In total, 288 OF samples were collected with cotton ropes and 216 with sponges and analysed for the biomarkers: cortisol, alpha-amylase, oxytocin (stress), haptoglobin (inflammation), procalcitonin (sepsis), adenosine deaminase, immunoglobulin G (immune system), ferric reducing antioxidant power (redox status), and creatine kinase, lactate dehydrogenase and total protein (general homeostasis). Samples were also scored visually for dirtiness using a score from 1 (clean) to 5 (very dirty). Results Rope-collected OF had higher levels of dirtiness (3.7 ± 0.04) compared to sponge-collected OF (2.7 ± 0.15) and had higher values than sponges for cortisol, procalcitonin, oxytocin, haptoglobin, total protein, lactate dehydrogenase and ferric reducing antioxidant power. All biomarkers decreased in value with age. Immunoglobulin G did not perform well for any of the two collection methods. Discussion and conclusion The results showed a clear effect of age on the biomarkers in OF collected with both, sponges or ropes. Sponges provided a cleaner sample than cotton ropes for biomarker analysis. Both methods are easy to apply under the commercial conditions in pig farms although sponges may take more time in early weaner stages. From a practical point of view, sampling with sponges achieved the best combination of reduced sampling time and low contamination

Changes in a Comprehensive Profile of Saliva Analytes in Fattening Pigs during a Complete Productive Cycle : A Longitudinal Study

The aim of this study was to evaluate whether a panel of 29 salivary biomarkers of stress, immunity, inflammation, redox homeostasis and other physiological functions can change in healthy fattening pigs when monitoring the different phases of their productive cycle and can be influenced by various sources of variations such as gender and performance parameters. Several analytes showed changes due to the productive cycle, with a majority of the analytes showing higher values at lactation and at the beginning of nursery. Additionally, differences were seen due to sex. These differences can be related in some cases with performance parameters and should be taken into consideration for an appropriate interpretation of the analytes. A comprehensive panel of 29 salivary analytes was measured in fattening pigs to evaluate its possible changes along their productive cycle. The identification of those changes would allow a better interpretation of the results according to the productive phase of the animal. Saliva samples were obtained from 49 Large-White pigs (24 females, 25 males) in suckling phase, at the beginning and the end of the nursery phase, and at the beginning and the end of the growing phase. Several analytes changed according to the phase of the productive cycle, with most of the analytes showing higher values at lactation and at the beginning of nursery. Additionally, differences were seen due to sex. When possible relations between performance parameters and analytes were evaluated, significant positive but weak relationships were found between weight at birth and salivary γ-glutamyl transferase, and between back-fat thickness and salivary lactate dehydrogenase. In conclusion, differences in the values of salivary analytes can be found in fattening pigs depending on the productive phase and sex of the animals

S-100 Proteins: Basics and Applications as Biomarkers in Animals with Special Focus on Calgranulins (S100A8, A9, and A12)

S100 proteins are a group of calcium-binding proteins which received this name because of their solubility in a 100% saturated solution of ammonium sulphate. They have a similar molecular mass of 10–12 KDa and share 25–65% similarity in their amino acid sequence. They are expressed in many tissues, and to date 25 different types of S100 proteins have been identified. This review aims to provide updated information about S100 proteins and their use as biomarkers in veterinary science, with special emphasis on the family of calgranulins that includes S100A8 (calgranulin A; myeloid-related protein 8, MRP8), S100A9 (calgranulin B; MRP14), and S100A12 (calgranulin C). The proteins SA100A8 and S100A9 can be linked, forming a heterodimer which is known as calprotectin. Calgranulins are related to the activation of inflammation and the immune system and increase in gastrointestinal diseases, inflammation and sepsis, immunomediated diseases, and obesity and endocrine disorders in different animal species. This review reflects the current knowledge about calgranulins in veterinary science, which should increase in the future to clarify their role in different diseases and potential as biomarkers and therapeutic targets, as well as the practical use of their measurement in non-invasive samples such as saliva or feces

Saliva Sampling Material Matters : Effects on the Results of Saliva Analysis in Pigs

There is an increasing interest in measuring biomarkers from saliva, as samples can be obtained noninvasively. This is particularly true in pigs where blood sampling is technically complicated and stressful. In pigs, studies regarding how the absorbent material used for collection affects analytical results are scarce. The aim of this study was to evaluate how the material used for saliva collection can affect the results of analyses for the different types of biomarkers that can be measured in the saliva of pigs. In this report, polypropylene sponges gave higher sample saliva volumes and, in some salivary analytes, provided different values compared with cotton rolls. Therefore, it can be concluded that the type of material used can influence the results of saliva analysis. The use of saliva as a biological sample from pigs is of high practical interest because blood collection from pigs is difficult and stressful. In this study, the influence of two different materials, a cotton roll and a polypropylene sponge, in porcine saliva collection was evaluated. For this purpose, the effect of the material used for sampling was evaluated in a panel of 13 analytes, including those related to stress (cortisol and oxytocin), inflammation and immunity (adenosine deaminase, haptoglobin and myeloperoxidase), redox homeostasis (the cupric reducing ability of saliva, the ferric reducing activity of saliva, and the Trolox equivalent antioxidant capacity), and sepsis (procalcitonin), as well as other routine analytes related to metabolism and different tissues and organs, such as lactate dehydrogenase, creatine kinase, urea, and total protein concentration. The polypropylene sponge provided a higher sample volume than the cotton roll. Although the results of some salivary analytes were equivalent for both materials, other analytes, such as creatine kinase, haptoglobin and total proteins, showed significant differences depending on the material used for saliva collection. Therefore, the type of material used for salivary collection in pigs should be considered when interpreting the results of analyses of the salivary analytes

Effects of pen faeces and feed contamination in biomarkers determination in oral fluid of pigs

peer-reviewedThe present study aims to evaluate the possible effects of the presence of pen faeces and feed on the measurement of a panel of biomarkers in porcine oral fluid. For this, clean porcine oral fluid was pooled and incubated with two different concentrations of pen faeces or feed representing a high or low level of contamination with each material. In addition, these pools were aliquoted and subjected to centrifugation, filtration or chemical clarification to evaluate if these techniques could revert the effects of those contaminants in biomarker evaluation. A panel of 21 biomarkers that assessed stress, inflammation, immune system and redox homeostasis among others, were measured for all aliquots. Changes of statistical relevance (p < 0.05) in oral fluid contaminated with pen faeces or feed versus untreated samples were observed for all methods employed with the exception of adenosine deaminase (ADA) and creatine kinase (CK) in oral fluid contaminated with pen faeces or feed. Pen faeces did not affect the measurement of haptoglobin, superoxide dismutase, CK, lactate dehydrogenase (LDH), ADA and cortisol (when the latter is measured by chemiluminescence); while uric acid, LDH, CK, ADA, and hydrogen peroxide methods were not affected by the presence of feed in oral fluid. The effects of centrifugation, filtration or chemical clarification with chitosan in these contaminated samples were modest and for most cases did not caused baseline levels on the measured biomarkers. In conclusion, the presence of pen faeces or feed in porcine oral fluid can interfere with the results obtained when analytes are measured

Comparison of the Effect of Two Different Handling Conditions at Slaughter in Saliva Analytes in Pigs

In this report, different handling conditions at slaughterhouse were studied to assess changes in salivary biomarkers. For this purpose, finishing pigs were divided into two groups, one in which handling was improved to minimize stress (Group A, n = 24, transported and stabled at the slaughterhouse at low density without mixing with unfamiliar animals throughout the whole process) and another one in which animals had a more stressful handling process (Group B, n = 24, transported and stabled at high density with unfamiliar animals). Saliva samples were taken the day before transport to the slaughterhouse at 8:00 a.m. (B0) and 12:00 a.m. (B4), and the day of slaughter just after unloading animals at the slaughterhouse at approximately 8:00 a.m. (S0) and after 4 h of lairage at approximately 12:00 a.m. (S4). Group B showed significantly higher cortisol, total esterase activity, oxytocin, adenosine deaminase and haptoglobin levels than the Group A at both S0 and S4 sampling times, and higher levels of calprotectin and creatine kinase at S4 sampling time. This report indicates that differences in the way in which the pigs are handled at the slaughterhouse can lead to changes in salivary biomarkers and opens the possibility of the use of biomarker at slaughter to monitor handling conditions