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Abstract Background Cortisone is derived from cortisol through the action of the enzyme 11β-hydroxysteroid dehydrogenase type II, and it has gained importance in recent years as a biomarker of stress. This study aimed to develop and validate an assay for the measurement of cortisone in pig saliva and evaluate whether its concentration varies in stressful situations. For this purpose, a specific immunoassay was developed and validated analytically, and a study was performed to evaluate whether cortisone concentrations in saliva can vary under heat stress conditions. Results The assay proved to be accurate, reliable, and sensitive for the measurement of cortisone in pig saliva. The limit of detection of the assay was set at 0.006 ng/ml, and the lower limit of quantification was 0.023 ng/ml. It also correlated significantly with the results obtained by LC‒MS/MS (P = 0.003; r = 0.64). In addition, the cortisone concentration in animals subjected to prolonged heat stress decreased significantly 15 days after treatment (P

Abstract Background Swine dysentery (SD) is a severe mucohaemorrhagic colitis in pigs caused classically by Brachyspira hyodysenteriae. Although several aspects of B. hyodysenteriae infection dynamic are already described, further research in the early stage of this infection is required. In this study, 7-week-old pigs were orally challenged with B. hyodysenteriae to obtain information about faecal shedding, macro and microscopic intestinal lesions and serum acute phase proteins in pigs at the onset of B. hyodysenteriae shedding (early infection group, n = 8), in pigs with mucohaemorrhagic diarrhoea (acute infection group, n = 8) and in non-infected controls (n = 16). Results First B. hyodysenteriae detection by q-PCR and first loose stools with blood and mucus occurred both at 8 days post-inoculation. The lapse between a positive q-PCR and observation of mucohaemorrhagic diarrhoea ranged from 0 to 3 days, except in a single pig in which this period lasted 5 days. Macroscopic lesions were observed in the large intestine from both infected groups although more frequent and severe in acute infection group. Microscopic observation of the apex mucosa revealed that in early infection only higher ulceration values were observed compared to healthy controls. In contrast, the acute infection group exhibited higher ulceration, neutrophils infiltration and increased mucosal thickness compared to the other two groups. Among the serum biomarkers tested, only haptoglobin, C-reactive protein, and creatine kinase showed a significant increase in pigs in the acute infection period compared to controls, whereas haptoglobin was the only factor with a significant increase at the early infection compared to non-infected animals. Conclusions This study provides new insights about SD and remarks the complex and limited options to perform an early detection of infected animals beyond PCR diagnosis.

Equine gastric ulcer syndrome (EGUS) is a common condition in horses. This study explores the use of liquid proteomics to identify new biomarkers in saliva and serum to monitor EGUS treatment. The proteomes of horses with EGUS before and after a successful treatment with omeprazole were analysed. In saliva, 503 proteins were identified, with 7 upregulated and 6 downregulated post-treatment. Among the proteins that changed, there was an increase in vimentin, linked to wound healing, and a decrease in podocalyxin, associated with tissue damage. In serum, 206 proteins were found, with significant changes in 5. Keratin type I increased, supporting epithelial integrity, whereas immunoglobulin lambda decreased, indicating a reduced immune response. Gene ontology analysis revealed a decrease in immune-related pathways after successful treatment. Overall, 13 proteins in saliva and 5 in serum showed significant changes after treatment, highlighting the differential responses of saliva and serum in EGUS. This report creates new avenues for discovering potential biomarkers to monitor EGUS treatment, which is of high importance for the management of this prevalent disease.

Objetivo. Estimar parámetros genéticos para peso a los ocho meses de edad (W8M), edad al primer parto (AFC) y primer intervalo entre partos (FCI) usando parentesco genómico y por pedigrí. Materiales y métodos. Se utilizaron 481, 3063 y 1098 registros fenotípicos para W8M, AFC y FCI, respectivamente. La información genómica estuvo compuesta por una población de 718 animales genotipados con un chip que incluyó 30106 marcadores genéticos tipo polimorfismo de nucleótido simple (SNP). Modelos univariado y bivariado fueron construidos bajo la metodología del mejor predictor lineal insesgado convencional (BLUP) y genómico en una etapa (ssGBLUP). Resultados. Las heredabilidades para W8M, AFC y FCI variaron desde 0.25 a 0.26, 0.20 a 0.22 y 0.04 a 0.08, respectivamente. Los modelos de AFC y FCI con la metodología ssGBLUP disminuyeron ligeramente el error y aumentaron la varianza genética aditiva, respectivamente. Conclusiones. La inclusión de información genómica mejora levemente la precisión de las estimaciones genéticas en esta población. Sin embargo, una población de animales genotipados más grande y con mayor conectividad genética por parentesco permitiría aumentar para los criadores el potencial de la metodología ssGBLUP en ganado Simmental de Colombia.

Metritis affects 5–20% of cows after parturition, negatively impacting animal welfare and the profitability of dairy farms, increasing culling rates and costs, and decreasing productivity and reproduction rates. This study compared the results of a comprehensive biochemical panel consisting of 25 salivary and 31 serum analytes between healthy cows (n = 16) and cows with metritis (n = 12). Descriptive parameters such as depression, rectal temperature, body condition score (BCS), heart rate, respiratory rate, mucous color, ruminal motility, vaginal discharge, milk production, and complete hematology analyses were also assessed for comparative purposes. The biochemistry analytes comprised five analytes related to stress, five to inflammation, five to oxidative status, and nineteen to general metabolism. The two-way ANOVA analysis revealed that, in saliva, eight biomarkers (lipase, adenosine deaminase (ADA), haptoglobin (Hp), total proteins, g-glutamyl transferase (gGT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and creatine kinase (CK)) were significant higher in cows with metritis. In serum, eight biomarkers (ADA, Hp, serum amyloid A (SAA), fibrinogen, ferritin, AOPPs/albumin ratio, non-esterified fatty acids (NEFAs), and bilirubin) were significantly higher in cows with metritis, whereas six (total esterase (TEA), albumin, urea, lactate, phosphorus, and calcium) were lower. Of the total number of 23 biomarkers that were measured in both saliva and serum, significant positive correlations between the two biofluids were found for six of them (Hp, FRAP, CUPRAC, AOPPs, urea, and phosphorus). Urea showed an R = 0.7, and the correlations of the other analytes were weak (R < 0.4). In conclusion, cows with metritis exhibited differences in biomarkers of stress, inflammation, cellular immune system, and general metabolism in both salivary and serum biochemistry profiles. These changes were of different magnitudes in the two biofluids. In addition, with the exception of ADA and Hp, the analytes that showed changes in the saliva and serum profiles of cows affected by metritis were different. Overall, this report opens a new window for the use of saliva as potential source of biomarkers in cows with metritis.

Abstract Background Leptospirosis is a neglected but widespread zoonotic disease throughout the world. Most mammals are hosts of Leptospira spp., including domestic cats, species in which no consensus has been reached on the clinical presentation or diagnosis of the disease. The study of acute-phase proteins (APPs) and biomarkers of oxidative status would contribute to knowledge about the disease in cats. This report evaluated four APPs: Serum amyloid A-SAA, Haptoglobin–Hp, albumin and Paraoxonase 1-PON1 and the antioxidant response through Total Antioxidant Capacity-TAC, in 32 free-roaming cats. Cats were classified as seroreactive for anti-leptospiral antibodies (group 1, n = 8), infected with Leptospira spp (group 2, n = 5) and leptospires-free cats (group 3, n = 19). Results SAA differences were observed between groups 1 and 2 (p-value = 0.01) and between groups 2 and 3 (p-value = 0.0001). Hp concentration differences were only detected between groups 2 and 3 (p-value = 0.001). Albumin concentrations only differed between groups 1 and 3 (p-value = 0.017) and 2 and 3 (p-value

Abstract Background 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.

Canine monocytic ehrlichiosis (CME), a vector-borne disease of worldwide distribution, causes coagulopathy in dogs. Thromboelastography-TEG measures the efficiency of coagulation. However, there is lack of knowledge about TEG evaluation in different stages of CME. Thus, this study aimed to evaluate the coagulation status and viscoelastic properties of blood using TEG parameters in dogs with naturally-occurring CME, and their potential to discriminate between subclinical and clinical forms of the disease. The relationship between TEG parameters and C-reactive protein (CRP) was also investigated. For these purposes, 29 E. canis-seropositive dogs were used (12 subclinical and 17 clinical forms), and 10 healthy dogs as controls. Kaolin-activated TEG was performed in all dogs. Platelets were lower (P

Abstract Background Streptococcus suis (S. suis) is a Gram-positive bacteria that infects pigs causing meningitis, arthritis, pneumonia, or endocarditis. This increases the mortality in pig farms deriving in severe economic losses. The use of saliva as a diagnostic fluid has various advantages compared to blood, especially in pigs. In this study, it was hypothesized that saliva could reflect changes in different biomarkers related to stress, inflammation, redox status, and muscle damage in pigs with S. suis infection and that changes in these biomarkers could be related to the severity of the disease. Results A total of 56 growing pigs from a farm were selected as infected pigs (n = 28) and healthy pigs (n = 28). Results showed increases in biomarkers related to stress (alpha-amylase and oxytocin), inflammation (haptoglobin, inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), total protein, S100A8-A9 and S100A12), redox status (advanced oxidation protein producs (AOPP)) and muscle damage (creatine kinase (CK), CK-MB, troponin I, lactate, aspartate aminotransferase, and lactate dehydrogenase). An increase in adenosine deaminase (ADA), procalcitonin, and aldolase in infected animals were also observed, as previously described. The grade of severity of the disease indicated a significant positive correlation with total protein concentrations, aspartate aminotransferase, aldolase, and AOPP. Conclusions This report revealed that S. suis infection caused variations in analytes related to stress, inflammation, redox status, and muscle damage in the saliva of pigs and these can be considered potential biomarkers for this disease.

Abstract Oxytocin has traditionally been known for its physiological effects on muscle contraction associated with birth and lactation, but in the last years is widely used as a biomarker of “positive experiences” in psychology and behavior. Different types of samples have been used for oxytocin measurements with saliva samples having the particular advantage of an easy and non-stressful collection. However, the low concentration of oxytocin in saliva can represent a limitation for its use. For this reason, sensitive assays and even a previous sample treatment in some cases are required for saliva oxytocin quantification. In addition, the lack of standardized and generally agreed-upon approach to peripheral oxytocin measurement leads to large discrepancies between different laboratories, that use different sample treatment protocols and different assays. The main objectives of this review are to describe the current status of the use of saliva for oxytocin measurement, provide details of the different sample processing techniques that can be applied and inform about the analytical techniques and assays available in different animal species, and also in humans for comparative purposes. It is expected that this information can contribute to an increase in the knowledge about the measurements of oxytocin in saliva and to its wider use in the future.

The objective of this study was to evaluate the possible use of spectrophotometric assays for the measurement of trace elements, including Zinc (Zn), Copper (Cu), Magnesium (Mg), and iron (Fe) in the saliva of horses and study their possible changes in equine gastric ulcer syndrome (EGUS). EGUS is a highly prevalent disease, with a current high incidence due to the increase in intensive management conditions. There are two EGUS diseases: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD), which can appear individually or together. For this purpose, automated spectrophotometric assays for measuring these analytes in horse saliva were analytically validated. Then, these analytes were measured in the saliva of horses with only ESGD, only EGGD, both ESGD and EGGD and a group of healthy horses. The methods used to measure the analytes were precise and accurate. Horses diagnosed with EGGD presented significantly lower levels of Zn and Mg. Fe concentrations were significantly lower in the saliva of horses with ESGD and EGGD. Overall, these results indicate that there are changes in trace elements in saliva in EGUS that could reflect the physiopathological mechanisms involved in this process and open the possibility of using trace elements as biomarkers of this syndrome.

Cystatin C, ammonia, and bicarbonate have been described to be biomarkers of sepsis and inflammation in humans. The saliva of pigs can be used to detect a wide range of pathogens but also many biomarkers that can be analyzed to evaluate different conditions such as stress (i.e., cortisol and alpha amylase), immune system (i.e., ADA, S100 proteins), inflammation (i.e., acute phase proteins), redox status (i.e., various antioxidants and oxidants), and general metabolism or the status of different organs and tissues. However, there is a lack of assays for the possible measurement and use of cystatin C, ammonia, and bicarbonate in saliva as biomarkers of sepsis or inflammation in pigs. The objective of this study was to validate commercially available automated assays for the measurement of cystatin C, ammonia, and bicarbonate in the saliva of pigs, having the advantage of using a noninvasive sample that is easy to collect. The assays were precise and accurate, and the recommended storage condition for the saliva samples was −80 °C. In addition, cystatin and ammonia showed significant increases in the saliva of pigs with S. suis infection, whereas bicarbonate decreased. Further studies would be recommended to increase knowledge about the possible potential applications of the measurements of these three analytes in the saliva of pigs as biomarkers to evaluate the animals’ health and welfare.

Abstract Background Hypothyroidism is the most common endocrine disorder diagnosed in dogs, leading to deleterious effects on a dog’s life quality. This study aims to evaluate changes in the redox status in canine hypothyroidism. For this purpose, a comprehensive panel of antioxidants and oxidants biomarkers were measured in serum and saliva of 23 dogs with hypothyroidism, 21 dogs with non-thyroidal illness, and 16 healthy dogs. Among the antioxidants, cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of plasma (FRAP), Trolox equivalent antioxidant capacity (TEAC), thiol, paraoxonase type 1 (PON-1) and glutathione peroxidase (GPx) were determined in serum and CUPRAC, ferric reducing ability of saliva (FRAS) and TEAC in saliva. The oxidant biomarkers included were total oxidant status (TOS), peroxide-activity (POX-Act), reactive oxygen-derived compounds (d-ROMs), advanced oxidation protein products (AOPP), and thiobarbituric acid reactive substances (TBARS) in serum and AOPP and TBARS in saliva. Results Results showed a significantly higher TEAC, PON-1, GPx, TOS, POX-Act, and d-ROMs, and a significantly lower AOPP in serum of dogs with hypothyroidism. Meanwhile, significantly lower FRAS and AOPP were observed in saliva of dogs with hypothyroidism. Once salivary concentrations were corrected based on their total protein concentrations, the only analyte showing significant changes was TBARS which was significantly higher in dogs with hypothyroidism. Conclusions Our results show that dogs with hypothyroidism present alterations in the redox status in both serum and saliva. This study should be considered a preliminary study and further research addressing these changes should be made using larger populations.

Heat stress accounts for millions of dollars in losses for swine producers worldwide. The aim of the present study was to determine and evaluate cortisol and cortisone in hair as indicators of thermal stress in growing pigs reared under high environmental temperatures. The study was carried out in two independent batches of commercial crosses of Lean Duroc and Pietrain in trials 1 and 2, respectively, during the growing period (from 40 to 100 kg; 81 days in trial 1 and 77 days in trial 2) in the same commercial farm in Spain during the summers of 2020 and 2021. In both cases, four rooms were used. In Trial 1, Room 1 had cooling and 11 pigs per pen; Room 2 had no cooling and 13 pigs per pen; Room 3 had no cooling and 11 pigs per pen, and Room 4 had cooling and 13 pigs per pen. In Trial 2, Rooms 2 and 3 had cooling and rooms 1 and 4 had no cooling, and all of them had 13 pigs per pen. Mean THI value was higher (p

Stress leukogram includes eosinopenia as one of its main markers (neutrophilia, eosinopenia, lymphopenia, and mild monocytosis). Cortisol is the main stress biomarker, which is also strongly correlated with the severity of gastrointestinal diseases. This study aimed to determine the relationship between salivary cortisol and the eosinophil cell count (EC) in equids with abdominal pain. To do this, 39 horses with abdominal pain referred to an emergency service were included. All samples were taken on admission, and several parameters and clinical data were included. Equids were classified according to the outcome as survivors and non-survivors. Non-surviving equids presented higher salivary cortisol concentrations (Non-Survivors: 1.580 ± 0.816 µg/dL; Survivors 0.988 ± 0.653 µg/dL; p < 0.05) and lower EC (Non-Survivors: 0.0000 × 103/µL (0.000/0.0075); Survivors: 0.0450 × 103/µL (0.010/0.1825); p < 0.01). In addition, the relationship between salivary cortisol concentration, EC, and the WBC was determined. Only a strong correlation (negative) was observed between cortisol and EC (r = −0.523, p < 0.01). Since cortisol is not an analyte that can be measured routinely in clinical settings such as emergencies, the EC could be a good alternative. While the results are promising, further studies are needed before EC can be used confidently in routine practice to predict survival in cases of abdominal pain.

Abstract Background The effect in a sialochemistry profile of the presence of usually available feed in dairy cows was evaluated by an in vitro experiment. For this purpose, a pooled clean saliva from five healthy dairy cows was incubated five times with a standard feed based on a total mixed ration (F), wheat hay (H), and grass (G). The salivary panel was integrated by biomarkers of stress (cortisol -sCor-, salivary alpha-amylase -sAA-, butyrylcholinesterase -BChE-, total esterase -TEA-, and lipase -Lip-), immunity (adenosine deaminase -ADA-), oxidative status (Trolox equivalent antioxidant capacity -TEAC-, the ferric reducing ability of saliva -FRAS-, the cupric reducing antioxidant capacity -CUPRAC-, uric acid, and advanced oxidation protein products -AOPP-), and enzymes, proteins, and minerals of general metabolism and markers of liver, muscle, and renal damage (aspartate aminotransferase -AST-, alanine aminotransferase -ALP-, γ-glutamyl transferase -gGT-, lactate dehydrogenase -LDH-, creatine kinase -CK-, creatinine, urea, triglycerides, glucose, lactate, total protein, phosphorus, and total calcium). Results Most of the evaluated analytes showed a coefficient of variations (CV) higher than 15% and/or significant changes compared with the clean saliva when feed was present. Some analytes, such as the oxidative status biomarkers (CV > 80%), AST (CV > 60%), or glucose (CV > 100%), showed significant changes with all the feed types tested. Others showed significant differences only with certain types of feed, such as LDH with F (CV > 60%) or triglycerides with F (CV > 100%) and H (CV > 95%). However, sCor or gGT remained unchanged (CV 0.05) in all the treatments. Conclusions The presence of feed can produce changes in most of the analytes measured in cows’ saliva, being of high importance to consider this factor when saliva is used as a sample to avoid errors in the interpretation of the results.

Abstract Background Procalcitonin (PCT) is a widely used biomarker of sepsis in human medicine and can have potential applications in the veterinary field. This study aimed to explore whether PCT could be measured in the saliva of pigs and whether its concentration changes in sepsis. Therefore, a specific assay was developed and analytically validated, and changes in PCT concentration were evaluated in two conditions: a) in an experimental model of sepsis produced by the administration of lipopolysaccharide (LPS) to pigs (n = 5), that was compared with a model of non-septic inflammation induced by turpentine oil (n = 4), and b) in healthy piglets (n = 11) compared to piglets with meningitis (n = 20), a disease that usually involves sepsis and whose treatment often requires large amounts of antibiotics in farms. Results The assay showed coefficients of variation within the recommended limits and adequate linearity after serial sample dilutions. The method's detection limit was set at 68 μg/L, and the lower limit of quantification was 414 μg/L. In the LPS experiment, higher concentrations of PCT were found after 24 h in the animals injected with LPS (mean = 5790 μg/L) compared to those treated with turpentine oil (mean = 2127 μg/L, P = 0.045). Also, animals with meningitis had higher concentrations of PCT (mean = 21515 μg/L) than healthy pigs (mean = 6096 μg/L, P value

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.

Abstract The use of saliva as a biological sample has many advantages, being especially relevant in pigs where the blood collection is highly stressful and painful, both for the animal and the staff in charge of the sampling. Currently one of the main uses of saliva is for diagnosis and detection of infectious diseases, but the saliva can also be used to measure biomarkers that can provide information of stress, inflammation, immune response and redox homeostasis. This review will be focused on the analytes that can be used for such evaluations. Emphasis will be given in providing data of practical use about their physiological basis, how they can be measured, and their interpretation. In addition, some general rules regarding sampling and saliva storage are provided and the concept of sialochemistry will be addressed. There is still a need for more data and knowledge for most of these biomarkers to optimize their use, application, and interpretation. However, this review provides updated data to illustrate that besides the detection of pathogens in saliva, additional interesting applicative information regarding pigs´ welfare and health can be obtained from this fluid. Information that can potentially be applied to other animal species as well as to humans.

Prenatal stress is the mechanism through which poor welfare of pregnant sows has detrimental effects on the health and resilience of their piglets. We compared two gestation housing systems (IMPROVED versus [conventional] CONTROL) in terms of sow stress and welfare indicators and sought to determine whether potential benefits to the sows would translate into improved offspring health. Sows were mixed into 12 stable groups (six groups per treatment, 20 sows per group) 29 days post-service in pens with free-access, full-length individual feeding/lying-stalls. CONTROL pens had fully slatted concrete floors, with two blocks of wood and two chains suspended in the group area. IMPROVED pens were the same but with rubber mats and manila rope in each stall, and straw provided in three racks in the group area. Saliva was collected from each sow on day 80 of pregnancy and analysed for haptoglobin. Hair cortisol was measured in late gestation. Sows’ right and left eyes were scored for tear staining in mid lactation and at weaning. Numbers of piglets born alive, dead, mummified, and total born were recorded. Piglets were weighed and scored for vitality and intra-uterine growth restriction (IUGR) at birth. Presence of diarrhoea in farrowing pens was scored every second day throughout the suckling period. IMPROVED sows had lower haptoglobin levels and tear-stain scores during lactation. IMPROVED sows produced fewer mummified piglets, and these had significantly lower IUGR scores, and scored lower for diarrhoea than piglets of CONTROL sows. Hence, improving sow welfare during gestation improved the health and performance of their offspring.

IntroductionFeline leukemia virus (FeLV) is a chronic disease that leads to the weakening of a cat's immune system. Platelet-rich plasma (PRP) offers therapeutic effects for multiple diseases, the use of PRP and growth factors (GFs) determination could be an alternative treatment to improve the quality of life in these patients. The objectives of this study were to determine and compare the concentration of platelets (PLTs), red blood cells (RBCs) and white blood cells (WBCs) between samples of whole blood (WB), PRP and platelet-poor plasma (PPP) fractions, and to evaluate the concentration of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor β1 (TGF-β1) in both fractions in FeLV cats using a PRGF®-Endoret® protocol previously standardized in this species.MethodsWB was collected from 11 asymptomatic FeLV-positive cats. PRP and PPP was obtained following PRGF®-Endoret® technology according to centrifugation at 265 g for 10 min. Cellular components, RBCs, WBCs, PLTs, and the PDGF-BB and TGF-β1 concentrations in PRP and PPP fractions were determined.ResultsPLT in the PRP fraction was statistically higher than WB and PPP fraction, with no statistical differences between WB and PPP. PLT concentration increased 1.4 times in PRP fraction compared to WB. Mean platelet volume (MPV) did not differ significantly between the WB, PRP, and PPP fractions. Compared to WB, the absolute numbers of RBCs and WBCs were decreased by 99% and more than 95% in the PRP and PPP fractions, respectively. TGF-ß1 concentrations increased in PRP vs. PPP, with no changes in PDGF-BB.DiscussionBased on the degree of PLT enrichment and the absence of RBCs and WBCs, this blood product could be classified as a Pure Platelet-Rich Plasma (P-PRP). The presence of GFs in PRP and PPP samples suggests that the PRGF®-Endoret® methodology is suitable for obtaining PRP in FeLV cats, despite future studies are necessary to optimize the technique, standardize the results and assess clinical efficacy.

Abstract Background Saliva is being increasingly used as a sample for measuring biomarkers in several species and shows a high potential of use to detect and monitor stress. The weaning and grouping in dairy calves are a particularly stressful time. Therefore, the objectives of this study were to evaluate a panel of antioxidant and oxidant biomarkers in the saliva of calves on the day of weaning (W0), 2 days after weaning or milk withdrawal (W + 2), and 4 days after grouping (G + 4). In addition, to verify if cortisol and oxytocin concentrations are related to the biomarkers measured. Results Salivary cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of saliva (FRAS), Trolox equivalent antioxidant capacity (TEAC), advanced oxidation protein products (AOPP), and ferrous oxidation-xylenol orange (FOX) were significantly higher (P

Abstract Background Postpartum dysgalactia syndrome (PDS) is associated with a significantly higher activation of the inflammatory and stress response at parturition than in the healthy sow. Therefore, reliable and possibly non-invasive biomarkers for substantial increases of inflammation are searched to support the PDS diagnosis. This report studies the possible changes of the inflammatory marker enzyme adenosine deaminase (ADA) in serum and saliva of 38 PDS positive sows (PDS+) and 38 healthy sows (PDS-). Sampling was performed every 24 h from 60 h before to 36 h after parturition. Isoenzyme 1 (ADA1) and isoenzyme 2 (ADA2), as well as total ADA (tADA), were measured and their statistical association with several serum and saliva biomarkers of inflammation and stress was investigated. Results Compared to a baseline (60 to 36h prepartum), salivary activities of ADA1, ADA2 and tADA increased significantly over time in both PDS+ and PDS- sows, reaching their peaks after parturition. In serum from PDS- sows, no changes were observed over time in either ADA1, ADA2 or tADA. In PDS+ sows, serum ADA2 activity decreased temporarily after parturition followed by a significant increase compared to baseline. ADA1, ADA2 and tADA were all significantly associated with several inflammatory biomarkers and ADA1 in serum was associated with serum cortisol. Although serum activity was higher in PDS+ than in PDS- sows, the differences were not statistically significant. Further, no difference was noted between the groups in the analyses of saliva. Conclusions Salivary ADA1 and ADA2 increased in all sows after parturition, potentially as a response to the postpartum inflammation. However, no difference in the activity of ADA1, ADA2 and tADA were found between PDS+ and PDS- sows indicating inability to diagnose PDS under the conditions described in this report.

IntroductionIntra-articular infiltration of plasma rich in growth factors (PRGF) and adipose mesenchymal stromal cells (AMSCs) are known to inhibit osteoarthritis progression. However, in severely affected patients, the treatment cannot reach the deeper layers of the articular cartilage; thus, its potential is limited. To overcome this limitation, intra-osseous infiltrations have been suggested. The purpose of this study is to assess the impact of intra-osseous infiltration therapies on serum biomarkers of osteoarthritis and to assess cartilage regeneration macroscopically.Materials and methodsA total of 80 rabbits were divided into four groups based on the intra-osseous treatment administered on the day of surgery: control, PRGF, AMSCs and a combination of PRGF + AMSCs. In addition, all groups received a single intra-articular administration of PRGF on the same day. Serum biomarker levels were measured before infiltration and 28-, 56-, and 84-days post infiltration, and macroscopical assessment was conducted at 56- and 84-days follow-up post infiltration.ResultsIn the PRGF + AMSCs group, significantly lower concentrations of hyaluronic acid and type II collagen cleavage neoepitope were recorded at all time points during the study, followed by PRGF, AMSCs and control groups. Regarding macroscopical assessment, lower scores were obtained in PRGF + AMSCs group at all study times.DiscussionThe results suggest that the combination of intra-articular PRGF with intra-osseous PRGF or AMSCs achieves better results in rabbits with acute chondral defects and that intra-osseous infiltration is a safe procedure.

The family of calgranulins includes S100A8 (calgranulin A), S100A9 (calgranulin B), which can appear as a heterodimer known as S100A8/A9 or calprotectin, and S100A12 (calgranulin C). These proteins are related to different inflammatory conditions, immune-mediated diseases, and sepsis and are considered biomarkers of potential interest. This study aims to evaluate if S100A8/A9 and A12 could change in pigs with diarrhea due to E. coli and to compare the changes of S100A8/A9 and A12 with other analytes in order to explore the possible causes or mechanisms involved. For this purpose, a panel integrated by analytes related to inflammation (haptoglobin, inter-alpha trypsin inhibitor 4 (ITIH4), and total protein); immune system (adenosine deaminase, ADA); stress (alpha-amylase); tissue damage (lactate and lactate dehydrogenase (LDH)); sepsis (aldolase) and redox status (ferric-reducing ability of saliva (FRAS) and advanced oxidation protein products (AOPP)) was evaluated. S100A8/A9 and A12 and the other analytes measured in this study showed increases in the saliva of pigs with diarrhea due to E. coli. S100A8/A9 and/or A12 showed a significant correlation of different magnitude with some of the other analytes evaluated. Further studies should be conducted to gain knowledge about the possible practical applications as biomarkers of the measurements of S100A8/A9 and A12 in the saliva of pigs.

This study is aimed at evaluating the effect of reducing stocking density and using cooling systems to mitigate the negative effects of high temperatures in growing pigs (females and castrated males) reared in intensive conditions (from 25 to 100 kg) during summer (June to October 2020). The experimental design was a 2 × 2 factorial where pigs were provided with an evaporative cooling system and/or raised at regular or at lower stocking densities (i.e., 0.68 to 0.80 m2/animal). Treatments were distributed in four different rooms containing sex-balanced pens with either castrated males or females. Temperature and humidity were recorded throughout the experiment, and the temperature–humidity index was calculated. Heat stress (HS) on pigs was measured through changes in animals’ performance, animal-based indicators (dirtiness and activity budget) and physiological indicators (neutrophil/lymphocyte ratio and hair cortisol). The use of cooling, lowering stocking density and the combination of both strategies had positive effects on pigs’ final body weight (+5 kg, +3 kg, +9 kg, respectively; p < 0.001). The prevalence of dirtiness was similar at the stocking densities tested, and no clear effect of the cooling system was found. Both mitigation strategies lowered the physiological indicators of stress, although only hair cortisone can be considered an indicator of HS. In conclusion, both mitigation strategies are effective in improving pig welfare and performance, especially when both are combined. The severity of the stocking density effect may depend on the severity of the temperature.

Canine obesity is the most common nutritional disorder and is associated with decreased quality of life and longevity as well as comorbidities including cardiorespiratory, endocrine, oncologic, or orthopaedic disorders. Ferritin is a major acute-phase protein in dogs, increasing during inflammation; however, it could also be affected by other conditions, including trauma, iron metabolism dysregulations, neoplasia, or hypoxia. Higher ferritin levels have been reported in obese humans, but ferritin has not been explored in canine obesity. To evaluate the possible changes in serum ferritin in canine obesity, ferritin levels from lean/normal weight (CG, n = 55) and overweight/obese dogs (OG, n = 37) were measured, together with complete hemogram and biochemical analyses. Statistically significant higher ferritin levels (1.2-fold) were found in OG (median, (interquartile range), 204 (166–227.5) µg/L) in comparison to CG animals (172 (137–210) µg/L)), with median levels of ferritin in OG dogs above the reference range for healthy animals in our laboratory (60–190 µg/L). In addition, statistically significant higher mean corpuscular volume (MCV), mean cell haemoglobin concentration (MCHC), total proteins, globulins, haptoglobin, total ferric fixation capacity (TIBC), alkaline phosphatase (ALP), butyrylcholinesterase (BChE), triglycerides, and calcium were observed in OG in comparison to CG. The higher levels in ferritin, together with higher TBIC, haematocrit, and MCV, could indicate tissue hypoxia in obese dogs.

Abstract Reactive oxygen species (ROS) are reactive compounds derived from oxygen. In biological systems, an excessive amount of ROS can cause oxidative damage to biological macromolecules being involved in different diseases. Several assays have been developed in the last 30 years for ROS evaluation. The objective of this article will be to provide an update about the spectrophotometric methods currently used in the assessment of ROS in serum. The chemical basis of four different techniques will be reviewed, and examples of their possible applications will be provided. A particular emphasis about the practical applications of these assays in the dog will be made, but selected information about their use in humans will also be presented for comparative purposes, following a One-Health approach. The information about the spectrophotometric assays presented in this paper should be interpreted with caution once limited information about them is available yet, and further studies should be performed to clarify what they measure and their clinical application. Ideally, when applied to evaluate a sample’s oxidative status, they should be incorporated in a panel of analytes where other oxidants, antioxidants, and biomarkers of inflammation were also included.

Abstract Background The possible use of oxytocin in saliva as an indicator of positive emotions in bovine species has been poorly investigated. In the present study, two new assays (one using a monoclonal antibody and the other using a polyclonal antibody) for the measurement of oxytocin in bovine saliva were developed and validated. Also, the changes in oxytocin in saliva were explored in two different situations. One was around parturition, and for this purpose, saliva samples from 13 cows were collected at three different times: 7 days before the parturition, the day of parturition and 7 days after the parturition. The second situation was weaning and grouping of calves, and for this purpose, saliva from 25 calves was collected at three different times: 1 day before weaning, 2 days after weaning or milk withdrawal and 4 days after grouping calves. Results In cows, oxytocin concentrations showed an increase on the day of parturition with both assays, while in calves, oxytocin concentrations showed a decrease 4 days after the grouping. Conclusions The assays validated in this report could be used for the measurement of oxytocin in bovine saliva and detect changes in this analyte that can occur in different physiological or productive situations such as parturition and weaning.

Equine gastric ulcer syndrome (EGUS) is a highly prevalent disease that affects horses worldwide. Within EGUS, two different forms have been described: equine squamous gastric disease (ESGD) and equine glandular gastric disease (EGGD). The associated clinical signs cause detrimental activity performance, reducing the quality of life of animals. Saliva can contain biomarkers for EGUS that could be potentially used as a complementary tool for diagnosis. The objective of this work was to evaluate the measurements of calprotectin (CALP) and aldolase in the saliva of horses as potential biomarkers of EGUS. For this purpose, automated assays for the quantification of these two proteins were analytically validated and applied for detecting EGUS in a total of 131 horses divided into 5 groups: healthy horses, ESGD, EGGD, combined ESGD and EGGD, and horses with other intestinal pathologies. The assays showed good precision and accuracy in analytical validation, and they were able to discriminate between horses with EGUS and healthy horses, especially in the case of CALP, although they did not show significant differences between horses with EGUS and horses with other diseases. In conclusion, salivary CALP and aldolase can be determined in the saliva of horses and further studies are warranted to elucidate the potential of these analytes as biomarkers in EGUS.

Background and Aim: Nitrate (NO3–) reduces enteric methane emissions and could be a source of non-protein nitrogen in ruminant feeds. Nonetheless, it has a potential toxic effect that could compromise animal health and production. The purpose of this study was to determine the effects of progressive inclusion of NO3– in the diet on the hematological, biochemical, and blood gases parameters, in turn, the effects on feed intake and live weight gain (LWG) in Holstein calves. Materials and Methods: Eighteen Holstein heifers and steers (nine animals/treatment) were maintained in individual pens for 45 days. Animals were randomly allocated to either a control or nitrate diet (ND) (containing 15 g of NO3–/kg of dry matter [DM]). The biochemical parameters and blood gases were analyzed only in the NO3– group on days: -1, 1, 7, 13, 19, and 25 corresponding to 0, 20, 40, 60, 80, and 100% of the total inclusion of NO3– in the diet, respectively. In addition, DM intake (DMI) and LWG were evaluated among dietary treatments. Results: Feeding the ND did not influence DMI or LWG (p>0.05). Methemoglobin (MetHb) and deoxyhemoglobin increased according to the NO3– concentrations in the diet (p0.05). However, glucose, urea, aspartate aminotransferase (AST), and retinol concentrations increased (p

In chickens, early life exposure to environmental microbes has long-lasting impacts on gastrointestinal (GI) microbiome development and host health and growth, via mechanisms that remain uncharacterized. In this study, we demonstrated that administrating a fecal microbiome transplant (FMT) from adults to day-of-hatch chicks results in significantly higher body mass of birds and decreased residual feed intake (RFI), implying enhanced feed efficiency, at 6 weeks of age. To assess the potential mechanisms through which FMT affects adult bird phenotype, we combined 16 S rRNA gene amplification, metagenomic, and comparative genomic approaches to survey the composition and predicted activities of the resident microbiome of various GI tract segments. Early life FMT exposure had a long-lasting significant effect on the microbial community composition and function of the ceca but not on other GI segments. Within the ceca of 6-week-old FMT birds, hydrogenotrophic microbial lineages and genes were most differentially enriched. The results suggest that thermodynamic regulation in the cecum, in this case via hydrogenotrophic methanogenic and sulfur-cycling lineages, potentially serving as hydrogen sinks, may enhance fermentative efficiency and dietary energy harvest capacity. Our study provides a specific mechanism of action through which early-life microbiome transplants modulate market-relevant phenotypes in poultry and, thereby, may represent a significant advance toward microbiome-focused sustainable agriculture.

IntroductionPlatelet-rich plasma (PRP) is an autologous plasma with platelet (PLT) concentration above that of whole blood (WB). PLTs contain growth factors (GFs) that promote tissular repair.ObjectivesTo determine and compare the concentrations of PLT, red blood cells (RBC) and white blood cells (WBC) between WB samples, PRP and platelet poor plasma (PPP) samples; and to analyze the concentrations of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor β1 (TGF-β1) in the PRP and PPP of healthy adult cats using a standardized protocol with PRGF®-Endoret® characteristics.Material and MethodsWB was collected from 30 cats. PRP was obtained following three centrifugation protocols using PRGF®-Endoret® technology: 255, 260, and 265 g for 10 min each. The cellular components, RBC, WBC, PLT, and the concentrations of PDGF-BB and TGF-β1 in the PRP and PPP fractions were determined for each protocol.ResultsPLTs in the PRP fraction were statistically higher than WB, with no statistical differences between PPP and WB. In PRP fraction, PLT concentration was increased 1.4 times on average at 255 g; 1.3 times at 260 g and, 1.5 times at 265 g without statistical differences among them. The mean platelet volume (MPV) was significantly higher in WB compared to PRP and PPP fractions without significant differences between protocols. Compared to WB, the number of RBCs and WBCs was reduced by 99% and by more than 95% in PRP and PPP respectively, without significant differences between protocols. PDGF-BB concentrations were statistically higher in PRP than in PPP fractions, however, TGF-ß1 concentrations did not vary between fractions at 260 g. Comparing the three protocols within PRP and PPP fractions, no differences in PDGF-BB and TGF-ß1 concentrations were observed.Clinical RelevanceThe study shows scientific evidence regarding the obtention of PRP in cats using the PRGF®-Endoret® technology for the quantification of PDGF-BB and TGF-ß1. At 265 g for 10 min, PLT concentration was increased 1.5 times with unnoticeable erythrocytes and leukocytes in the samples. These results clearly show that the PRGF®-Endoret® methodology is suitable to obtain PRP in cats. Further studies are needed to determine the clinical efficacy of the obtained PGRF in the treatment of different pathologies in cats.

Abstract Background Measurement of adenosine deaminase (ADA) can provide information about cell-mediated immunity. This report’s objective was to study the enzymatic activity of total ADA (tADA) and its isoenzymes ADA1 and ADA2 in canine, equine, porcine, and bovine serum and saliva and their changes in different inflammatory situations in each species. Besides, an automated method for ADA2 measurement was developed and validated. Results tADA was present in serum and saliva of healthy animals of the four species. Erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) concentration of 0.47 mM was needed for ADA1 inhibition in canine and porcine samples (serum and saliva) and bovine saliva, whereas for equine saliva 0.94 mM was needed. ADA2 activity was not detected in bovine serum and was very low or absent in equine serum and bovine saliva. An automated procedure to measure ADA2 consisting of adding EHNA to a commercial reagent for tADA measurement provided repetitive (coefficients of variation 0.90) results, being equivalent to a manual incubation of the sample with EHNA at a similar concentration. Salivary tADA, as well as ADA1 and ADA2, were higher in dogs with leishmaniosis, horses with acute abdominal disease and pigs with lameness than in healthy animals. tADA and isoenzymes in saliva showed a positive significant correlation with serum ferritin in dogs (r = 0.602, P

Abstract Background Captive and free-ranging wild mammals have been recognized as potential reservoirs of Leishmania infantum infection. The aim of this study was to describe the first clinical case of leishmaniosis in the Eurasian otter. Case presentation A case of clinical leishmaniosis is reported in a 4-year-old male Eurasian otter housed at a wildlife park (Murcia, South Eastern Spain). The Eurasian otter showed bilateral epistaxis, anorexia, apathy, and weight loss. A complete blood cell count and biochemical analyses revealed hyperproteinemia, hyperglobulinemia, decreases of paraoxonase-1, increases of haptoglobin and ferritin, and proteinuria. Bilateral nephropathy with hydronephrosis, mesenteric lymphadenomegaly, and ascites were also observed. L. infantum infection was confirmed by microscopy (amastigotes were detected in macrophages from spleen aspirate), molecular diagnosis (L. infantum DNA was detected by real-time polymerase chain reaction), and serology (anti-Leishmania IgG2 antibodies were detected by time-resolved immunofluorometry). The animal was treated with allopurinol for 3 months and gained weight, the epistaxis disappeared, and the ferritin concentration decreased. Conclusions This is the first report of clinical leishmaniosis in the Eurasian otter. Our results suggest that Eurasian otters are susceptible to infection with L. infantum and can develop clinical leishmaniosis in endemic areas.

La pasta de higuerilla (Ricinus communis L) contiene sustancias altamente tóxicas, por lo que se evaluó la efectividad de tres métodos de desintoxicación y su inclusión en dietas para pollos de engorda. Se evaluaron cinco tratamientos (dietas experimentales): dieta testigo a base de maíz y pasta de soya (PS), pasta de higuerilla sin desintoxicar (PH), pasta de higuerilla tratada con autoclave (PHA), pasta de higuerilla tratada con el método químico (PHQ) y pasta de higuerilla tratada con autoclave y método químico (PHAQ). Cada tratamiento se asignó al azar a siete unidades experimentales con 10 pollos cada una. Las variables evaluadas fueron: consumo de alimento (COA), conversión alimenticia (CA), ganancia de peso (GP) rendimiento en canal (RC), rendimiento de pechuga (RP), rendimiento de pierna con muslo (RPM), desarrollo del sistema digestivo, habilidad para caminar (HC), angulación valgus /varus (AVV) y latencia a postrarse (LP). Los pollos alimentados con PH y PHMQ tuvieron menor COA y GP (P0.05). Los resultados evidencian que el tratamiento con autoclave (1 atmosfera de presión durante 60 min a 121 °C) disminuyó la toxicidad en la pasta de higuerilla, ya que las aves del tratamiento PHMA tuvieron un comportamiento productivo similar (P>0.05) a las de la dieta testigo.

Tres protocolos reproductivos fueron evaluados: 1) PG: inyección de PGF2α en el día 50 posparto e inseminación (IA) en base a detección del estro. 2) OVS (Ovsynch: día 0, GnRH; día 7, PGF2α; día 9, GnRH; día 10, IA); y 3) ROV (GnRH + Ovsynch: día- 7, GnRH; día 0, GnRH; día 7, PGF2α; día 9, GnRH; día 10, IA). Además, el efecto de la somatotropina (bST) a la IA, sobre la fertilidad al primer servicio posparto (FERT), y la tasa de preñez a 99 días posparto (PP). FERT fue similar en ROV y OVS (36.2 vs 36.6 %) (P>0.05); y mayor a PG (27.3 %) (P0.05). PG y sin bST (22.5 %) fue menor que OVS con (38.5 %), y sin bST (33.7 %); y que ROV con bST (37.0 %) y sin bST (36.1 %); y PG con bST (32.9 %) (P

Assisted reproductive technologies play a major role in the cattle industry. An increase in the use of in vitro-derived embryos is currently being seen around the globe. But the efficiency and quality of the in vitro-derived embryos are substandard when compared to the in vivo production. Different protocols have been designed to overcome this issue, one of those being the use of reproductive fluids as supplementation to embryo culture media. In this study, in vitro-derived calves produced with reproductive fluids added to their embryo production protocol were followed for the first year of life pairwise with their in vivo control, produced by artificial insemination (AI), and their in vitro control, produced with standard supplementation in embryo production. The objective was to assess if any differences could be found in terms of growth and development as well as hematological and biochemical analytes between the different systems. All the analysed variables (physical, hematological, and biochemical) were within physiological range and very similar between calves throughout the entire experiment. However, differences were more evident between calves derived from standard in vitro production and AI. We concluded that the use of reproductive fluids as a supplementation to the embryo culture media results in calves with closer growth and development patterns to those born by AI than the use of bovine serum albumin as supplementation.

Calprotectin (CALP, S100A8/A9), also named myeloid-related protein 8/14, is a dimer complex of S100A8 and S100A9 that belongs to the S-100 protein family. It is involved in inflammation and has a wide range of proinflammatory functions, such as cytokine production and regulation of leukocyte adhesion, migration, and phagocytosis. In humans, CALP traditionally can be measured in faeces, serum, and saliva as a biomarker of inflammation and sepsis. The objective of this study was to validate an automated assay for CALP measurements in the saliva of pigs, having the advantage of the use of a non-invasive sample that is easy to collect. The assay was precise and accurate. CALP in saliva measured by this assay showed significant changes depending on the hour of the day. It also showed significant increases in the saliva of pigs after the administration of lipopolysaccharide (LPS), and showed a rise, although with increases of lower magnitude, after a stressful stimulus. Further studies should be made to gain knowledge about the possible practical applications of the measurements of CALP in the saliva of pigs as a biomarker to evaluate the animals’ health and welfare.

The objective of the present study was to measure the concentration of Paraoxonase-1 (PON-1) and N-terminal-prohormone-B-type natriuretic peptide (NT-proBNP), in the serum of dogs with degenerative Mitral Valve Disease (MVD), in order to identify their association with the clinical stage and specific clinico-pathologic and echocardiographic findings.Eighty dogs diagnosed with MVD and staged according to the ACVIM (American College of Veterinary Internal Medicine) consensus statement (B1, B2, C and D), based on their clinical, radiographic, and echocardiographic findings, were included in the study. NT-proBNP was measured only in stage B1 and B2 dogs. Clinical stage did not have a significant effect on PON-1 concentrations (p = 0.149), but NT-proBNP levels were lower in B1 dogs (p = 0.001). A significant correlation between PON-1 and total plasma proteins (p = 0.001), albumin (p = 0.003) and white blood cell count (p = 0.041) was detected, whereas there was no significant correlation (p = 0.847) between PON-1 and NT-proBNP concentrations. PON-1 showed a significant but weak negative correlation with normalized left ventricular internal diameter at diastole (LVIDdn) (p = 0.022) and systole (LVIDsn) (p = 0.012), as well as mitral valve E to A wave velocity ratio (MV E/A) (p = 0.015), but not with Left Atrial to Aortic root ratio (LA/Ao) (p = 0.892) or fractional shortening (FS%) (p = 0.944). PON-1 seems to be an insensitive marker of clinical stage and disease severity in MVD, but can be indicative of some clinico-pathological and echocardiographic changes. NT-proBNP changes are independent of oxidative stress.

In this report, the measurement of salivary biomarkers as an aid for diagnosis of equine gastric ulcer syndrome (EGUS) was studied. A comprehensive panel of 23 salivary analytes was measured in the saliva of horses affected by EGUS and compared to healthy animals and horses with other diseases clinically similar to EGUS but with a negative diagnosis at gastroscopic examination. A total of 147 horses were included in the study and divided into heathy population (n = 12), the EGUS group (n = 110), and the group of horses with other diseases (n = 25). From the 23 analytes studied, 17 showed increased values in EGUS horses when compared to healthy ones, and uric acid, triglycerides, and calcium were significantly increased in horses with EGUS compared to the group of other diseases. The receiver operating characteristic curve analyses showed a modest but significant discriminatory power of those three analytes to identify EGUS from other diseases with similar symptoms. The discriminatory power enhanced when the results of the three analytes were combined. In conclusion, the results showed that selected salivary analytes could have potential use as biomarkers in horses with EGUS.

Abstract This manuscript provides updated knowledge and some ideas and reflections about three areas which are currently of interest in the field of the animal laboratory science. These areas are the study of acute phase proteins (APPs) as biomarkers of inflammation, the use of saliva as a non-invasive sample for analyte measurements, and the development of education in the field of laboratory medicine. In the APPs, a seven-point plan for their interpretation in all veterinary species containing updated knowledge and future perspectives is described. Regarding the saliva, general concepts, examples of practical applications and the limitations and points to improve for the use of this fluid are explained. Finally, the recent evolution, current situation and possible ideas for future development of education in this field are commented. In addition to review the knowledge in these three specific areas, this report can help to provide a wide vision of the potential and future perspectives in veterinary laboratory science.

Abstract Background The biochemical components of saliva can change in certain pathologies in horses, for example in acute abdominal disease. The aim of this study was (1) to evaluate if a panel of biochemical analytes usually used in serum can be measured in saliva of horses and (2) to study the possible changes of these biochemical analytes in saliva of horses affected by acute abdominal disease. A panel of 23 analytes was analytically validated in saliva of horses and possible changes in these analytes in a pilot study with six healthy horses and six horses with acute abdominal disease were evaluated. The analytes with significant changes were then evaluated in a larger population of 20 healthy and 37 diseased horses. Results Seven analytes showed significant increases in the pilot study which were confirmed in the larger population. The analytes which showed significant changes, and their median fold increase and significance shown in the larger population were salivary γ-glutamyl transferase (gGT, 2.3 fold, P = 0.001), creatine kinase (CK, 6.2 fold, P

Abstract Background Biomarkers of oxidative stress in pigs have been measured in serum/plasma samples. However, blood collection in pigs can be highly stressful to the animals. Saliva is a biological fluid with several advantages in pigs over blood, since it can be easily collected without stress to the animals, being therefore an ideal sample in this species. The objective of this study was the validation of assays for the evaluation of oxidative stress status in saliva of pigs. For this purpose, three assays commonly used to evaluate the total antioxidant capacity (TAC): trolox equivalent antioxidant capacity (TEAC), cupric reducing antioxidant capacity (CUPRAC), and ferric reducing ability of plasma (FRAP)), one individual antioxidant (uric acid) and two assays to evaluate oxidant concentrations (advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2)) were measured and validated in porcine saliva. In addition, the possible changes of these assays in sows’ saliva during lactation were be studied. Results The methods had intra- and inter-assays coefficient of variation lower than 15%. They also showed an adequate linearity and recovery, and their detection limits were low enough to detect the analytes in saliva of pigs. Overall the analytical validation tests showed that the assays used in our study are valid and reliable for the evaluation of oxidative stress in porcine saliva. In addition, it was observed that these salivary biomarkers can change in a situation of oxidative stress such as lactation in sows. Conclusions All assays for salivary biomarkers of oxidative stress evaluated in this study have demonstrated a high analytical accuracy and low imprecision. In addition, it has been observed that these biomarkers showed significant changes in a situation of oxidative stress such as lactation in sows. Therefore, this study opens a new possibility of using saliva as a non-invasive sample to evaluate oxidative stress in pigs.

El objetivo de este estudio fue estimar parámetros genéticos con y sin la inclusión de parentesco genómico para la producción de leche acumulada a 60 (PL60), 150 (PL150), 210 (PL210) y 305 días (PL305) en ganado Simmental en Colombia. Un total de 2883 controles lecheros en 620 vacas de primer parto fueron utilizados. La información genómica se obtuvo a partir de 718 animales genotipados con un chip de una densidad de 30106 marcadores genéticos tipo polimorfismo de nucleótido simple (SNP). Se construyeron modelos de tipo univariado y bivariado bajo la metodología del mejor predictor lineal insesgado (BLUP) y genómico en una etapa (ssGBLUP). Los valores de heredabilidades para PL60, PL150, PL210 y PL305 variaron entre 0,20 a 0,27; 0,25 a 052; 0,30 a 0,35 y 0,20 a 0,23; respectivamente. La inclusión de parentesco genómico no aumentó las heredabilidades y tampoco la precisión de las estimaciones para las características asociadas a producción de leche. La escasez de información fenotípica y la baja conectividad genética entre la población genotipada y no genotipada podrían limitar procesos de selección genética para producción de leche a través del ssGBLUP en la población de ganado Simmental de Colombia.

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.

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.

La hormona del crecimiento bovina (bST) incrementa las con-centraciones séricas del Factor de Crecimiento Insulinoide tipo I (IGF-I). Estas hormonas favorecen el desarrollo folicular, la fer-tilización y la sobrevivencia embrionaria. La administración de Gonadotropina Coriónica Equina (eCG) al momento de retirar la esponja con Acetato de Fluorogestona (FGA), este progestáge-no aumenta la tasa de ovulación y la respuesta estral. En este estudio se probó la administración de bST 5 días (d) antes de re-tirar la esponja y eCG al retirar el tratamiento mejora la respuesta estral, la fertilidad y prolificidad en ovejas Pelibuey. Se utilizaron 100 ovejas Pelibuey con diferente número de partos. A todas las ovejas se les insertó una esponja intravaginal impregnada con 45 miligramos (mg) de FGA durante 12 d, 25 ovejas asignadas al azar fueron distribuidas en cuatro tratamientos: 1) bST ecibieron 125 mg de bST vía subcutánea (SC) cinco d antes de retirar la esponja con FGA; 2) eCG recibieron 300 Unidades Internacio-nales (UI) de eCG vía SC al momento de retirar la esponja; 3) bST+eCG recibieron 125 mg de bST vía SC cinco d antes de reti-rar la esponja de FGA y 300 UI eCG vía Intramuscular (IM) al mo-mento de retirar la esponja con FGA; 4) testigo o recibieron bST ni eCG. Se comparó la proporción de ovejas que presentaron estro, la Fertilidad y la Prolificidad. La respuesta estral fue similar (P>0,05) entre los grupos. El porcentaje de concepción fue ma-yor (P0,05) en los grupos eCG (66,%) y testigo (65,2%). La proporción de ovejas con parto múlti-ple fue similar (P>0,05) en los grupos bST (35%) y testigo (20%); sin embargo, fue mayor en los grupos eCG (69%) y eCG+bST (80%). Se concluye que el tratamiento con bST cinco d antes de retirar la esponja de FGA más eCG al momento de retirar la es-ponja incrementa la fertilidad y la prolificidad en ovejas Pelibuey

Changes in the salivary proteome in 12 horses with the two diseases included in equine gastric ulcer syndrome (EGUS), equine glandular gastric disease (EGGD) (n = 6) and equine squamous gastric disease (ESGD) (n = 6), were evaluated using a high-resolution LC-MS/MS analysis of TMT-labelled peptides and compared to 10 healthy control horses. Serum was also analysed for comparative purposes. The comparison between the horses with EGGD and controls showed significant changes in 10 salivary proteins, whereas 36 salivary proteins were differently abundant between ESGD and control groups. The most upregulated proteins in the case of EGGD were related to immune activation whereas, in horses with ESGD, the most significantly changed proteins were associated with squamous cell regulation and growth. Compared to serum, saliva showed a higher number of proteins with significant changes and a different pattern of changes. The proteins identified in our study, in addition to providing new information about the pathophysiological mechanisms in these diseases, could have the potential to be novel biomarkers for the diagnosis or monitoring of EGGD and ESGD.