DigitalUM :: Browsing by browse.metadata.contributordepartment "Medicina y Cirugía Animal"

Browsing by browse.metadata.contributordepartment "Medicina y Cirugía Animal"

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Embargo
A new highly sensitive immunoassay for the detection of adiponectin in serum and saliva of dogs and its application in obesity and canine leishmaniosis
(2019-07-31) Muñoz-Prieto, Alberto; Martínez-Subiela, Silvia; Cerón, José Joaquín; Tvarijonaviciute, Asta; Medicina y Cirugía Animal
Adiponectin is an adipokine that exerts insulin-sensitizing and antiinflammatory properties. The aim of this study was to develop and validate a new heterologous ultrasensitive assay based on amplified luminescent technology for adiponectin determination in serum and saliva of dogs. A complete analytical validation of the assay was made in these fluids, and also this assay was applied to quantify adiponectin in serum and saliva of obese and lean dogs and dogs with leishmaniosis. These conditions were selected because in obesity there is a controversy about how adiponectin concentrations change in dogs, and in case of canine leishmaniosis, although it is described a decrease in serum adiponectin, there are not studies about how adiponectin changes after treatment. A total of 11 dogs were used in the validation and 26 dogs with different body condition and 8 with canine leishmaniosis were used for the clinical evaluation of the new assay for adiponectin quantification in serum and saliva of dogs. The analytical evaluation showed that the developed method could measure adiponectin in serum and saliva of dogs with high repeatability and sensitivity, adding a limit of quantification lower than commercially available ELISAs. In addition, significantly higher adiponectin concentrations were found in lean dogs and a correlation between serum and saliva was observed (P < .01). Moreover, dogs with leishmania presented reduced levels of adiponectin in serum. In conclusion, a new assay has been developed for adiponectin measurements which is more sensitive and faster than the traditional ELISA assays requiring less sample volume.
Open Access
A Proteomic Approach to Elucidate the Changes in Saliva and Serum Proteins of Pigs with Septic and Non-Septic Inflammation
(MDPI, 2022-06-16) López Martínez, María José; Cerón, José J.; Ortin Bustillo, Alba; Escribano, Damián; Kuleš, Josipa; Beletić, Anđelo; Rubić, Ivana; González Sánchez, Juan Carlos; Mrljak, Vladimir; Martínez Subiela, Silvia; Muñoz Prieto, Alberto; Medicina y Cirugía Animal
Sepsis is a systemic inflammatory response triggered by an infectious agent and is recognized by the World Health Organization as a global concern, since it is one of the major causes of severe illness in humans and animals. The study of the changes that can occur in saliva and serum in sepsis can contribute to a better understanding of the pathophysiological mechanisms involved in the process and also to discover potential biomarkers that can help in its diagnosis and monitoring. The objective of this study was to characterize the changes that occur in the salivary and serum proteome of pigs with experimentally-induced sepsis. The study included five pigs with sepsis induced by LPS administration and five pigs with non-septic inflammation induced by turpentine for comparative purposes. In saliva, there were eighteen salivary proteins differentially expressed in the sepsis condition and nine in non-septic inflammation. Among these, significant increments in aldolase A and serpin B12 only occurred in the sepsis model. Changes in aldolase A were validated in a larger population of pigs with sepsis due to Streptococcus suis infection. In serum, there were 30 proteins differentially expressed in sepsis group and 26 proteins in the non-septic group, and most of the proteins that changed in both groups were related to non-specific inflammation. In the saliva of the septic animals there were some specific pathways activated, such as the organonitrogen compound metabolic process and lipid transport, whereas, in the serum, one of the main activated pathways was the regulation of protein secretion. Overall, saliva and serum showed different proteome variations in response to septic inflammation and could provide complementary information about the pathophysiological mechanisms occurring in this condition. Additionally, salivary aldolase A could be a potential biomarker of sepsis in pigs that should be confirmed in a larger population
Open Access
A Systematic Review and Meta-Analysis of Serum Adiponectin Measurements in the Framework of Dog Obesity
(2020-09-14) Muñoz-Prieto, Alberto; Cerón, José Joaquín; Martínez-Subiela, Silvia; Mrljak, Vladimir; Tvarijonaviciute, Asta; Medicina y Cirugía Animal
Adiponectin is an abundant plasma protein that is closely related to obesity and obesity-related pathologies. The molecule can be found in three different isoforms, each with different biological activities. Studies on canine obesity have suggested that adiponectin concentrations are decreased in obesity; however, no canine meta-analyses have been performed that feature all the required data. The aim of this study is to perform a systematic review and meta-analysis of studies that pertain to total and high molecular weight (HMW) adiponectin in relation to canine obesity. From 20 different studies, a total of 366 dogs with obesity and 349 normal weight dogs are included in the meta-analysis. Client-owned dogs were most represented, accounting for 54.3% of the dogs used, while experimental dogs enrolled in the studies made up the remaining 45.7%. The concentrations of total adiponectin in dogs with obesity were significantly lower compared with normal weight dogs. Additionally, adiponectin concentrations were significantly higher in dogs after a successful weight loss protocol compared to the start of the protocol and were significantly lower in dogs after gaining weight. In conclusion, although caution should be taken due to the relatively low number of studies that exist and the high heterogeneity between them, this meta-analysis indicates that adiponectin is decreased in obese dogs.
Embargo
Active paraoxonase 1 is synthesised throughout the internal boar genital organs
(BioScientifica, 2017-09) Barranco, Isabel; Pérez-Patiño, Cristina; Tvarijonaviciute, Asta; Parrilla, Inmaculada; Vicente-Carrillo, Alejandro; Álvarez-Rodríguez, Manuel; Cerón, José J.; Martínez, Emilio A.; Rodríguez-Martínez, Heriberto; Roca, Jordi; Medicina y Cirugía Animal
The paraoxonase type 1 (PON1) is an enzyme with antioxidant properties recently identified in the seminal plasma (SP) of several species, including the porcine. The aims of the present study were to (1) describe the immunohistochemical localisation of PON1 in the genital organs of fertile boars and (2) evaluate the relationship among PON1 activity and high-density lipoprotein cholesterol (HDL-C) concentration in fluids of the boar genital organs. Immunohistochemical analysis demonstrated that PON1 was present in testis (specifically in Leydig cells, blood vessels, spermatogonia and elongated spermatids), epididymis (specifically in the cytoplasm of the principal epithelial cells, luminal secretion and in the surrounding smooth muscle) and the lining epithelia of the accessory sexual glands (cytoplasmic location in the prostate and membranous in the seminal vesicle and bulbourethral glands). The Western blotting analysis confirmed the presence of PON1 in all boar genital organs, showing in all of them a band of 51 kDa and an extra band of 45 kDa only in seminal vesicles. PON1 showed higher activity levels in epididymal fluid than those in SP of the entire ejaculate or of specific ejaculate portions. A highly positive relationship between PON1 activity and HDL-C concentration was found in all genital fluids. In sum, all boar genital organs contributing to sperm-accompanying fluid/s were able to express PON1, whose activity in these genital fluids is highly dependent on the variable HDL-C concentration present.
Open Access
Aldose reductase B1 in pig seminal plasma: identification, localization in reproductive tissues, and relationship with quality and sperm preservation
(Frontiers Media, 2021-06-08) Mateo Otero, Yentel; Viñolas Vergés, Estel; Llavanera, Marc; Ribas Maynou, Jordi; Roca, Jordi; Yeste, Marc; Barranco, Isabel; Medicina y Cirugía Animal
Aldose reductase B1 (AKR1B1), a NADPH-dependent enzyme that belongs to the aldo-keto reductase protein superfamily, has been reported to be involved in both male and female reproductive physiology. The objectives of this study were: (1) to evaluate the concentration of SP-AKR1B1 in pig ejaculate fractions; (2) to describe the immunohistochemical localization of AKR1B1 alongside the boar genital tract; (3) to evaluate the relationship between SP-AKR1B1 and sperm quality/functionality parameters. Ejaculates from seven boars (one ejaculate per boar) were collected in separate portions [the first 10 mL of the sperm rich fraction (SRF-P1), the rest of the SRF (SRF-P2), and the post-SRF (PSRF)], and the concentration of SP-AKR1B1 was assessed using an enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry and immunoblotting targeting was conducted in the reproductive tissues of these boars. Additionally, the entire ejaculates of 14 boars (one ejaculate per boar) were collected and split into three separate aliquots for: (i) SP-AKR1B1 quantification; (ii) assessment of sperm concentration and morphology; and (iii) evaluation of sperm quality and functionality parameters upon ejaculate collection (0 h) and after 72 h of liquid storage at 17°C. Concentration of AKR1B1 in the SP of SRF-P1 (458.2 ± 116.33 ng/mL) was lower (P < 0.05) than that of SRF-P2 (1105.0 ± 229.80 ng/mL) and PSRF (1342.4 ± 260.18 ng/mL). Monomeric and dimeric AKR1B1 forms were expressed alongside the reproductive tissues, except in the bulbourethral glands. No relationship between SP-AKR1B1 and sperm quality/functionality parameters was observed either at 0 h or after 72 h of storage at 17°C. In conclusion, AKR1B1 is expressed in the reproductive organs of boars (except bulbourethral glands) and a higher concentration is found in the PSRF suggesting that seminal vesicles would be the main secretory source. However, this enzyme does not appear to be related to sperm quality/functionality or to the sperm ability to withstand liquid storage at 17°C.
Open Access
An Upgrade on the Rabbit Model of Anthracycline-Induced Cardiomyopathy: Shorter Protocol, Reduced Mortality, and Higher Incidence of Overt Dilated Cardiomyopathy
(2015-12-01) Talavera López, Jesús; Giraldo, Alejandro; Fernández del Palacio, María Josefa; García-Nicolás, Obdulio; Seva, Juan; Brooks, Gavin; Moraleda, Jose María; Medicina y Cirugía Animal
Current protocols of anthracycline-induced cardiomyopathy in rabbits present with high premature mortality and nephrotoxicity, thus rendering them unsuitable for studies requiring long-term functional evaluation of myocardial function (e.g., stem cell therapy). We compared two previously described protocols to an in-house developed protocol in three groups: Group DOX2 received doxorubicin 2 mg/kg/week (8 weeks); Group DAU3 received daunorubicin 3 mg/kg/week (10 weeks); and Group DAU4 received daunorubicin 4mg/kg/week (6 weeks). A cohort of rabbits received saline (control). Results of blood tests, cardiac troponin I, echocardiography, and histopathology were analysed. Whilst DOX2 and DAU3 rabbits showed high premature mortality (50% and 33%, resp.), DAU4 rabbits showed 7.6% premature mortality. None of DOX2 rabbits developed overt dilated cardiomyopathy; 66% of DAU3 rabbits developed overt dilated cardiomyopathy and quickly progressed to severe congestive heart failure. Interestingly, 92% of DAU4 rabbits showed overt dilated cardiomyopathy and 67% developed congestive heart failure exhibiting stable disease. DOX2 and DAU3 rabbits showed alterations of renal function, with DAU3 also exhibiting hepatic function compromise. Thus, a shortened protocol of anthracycline-induced cardiomyopathy as in DAU4 group results in high incidence of overt dilated cardiomyopathy, which insidiously progressed to congestive heart failure, associated to reduced systemic compromise and very low premature mortality.
Open Access
Analytical characterization of trace elements (zinc, copper, cadmium, lead and selenium) in saliva of pigs under common pathological conditions in the field: a pilot study
(BMC [Commercial Publisher], 2020) Sánchez, Jorge; Montilla, Miguel; Gutiérrez-Panizo, Cándido; Sotillo, Juan; Fuentes, Pablo; Montes, Ana; Gutiérrez, Ana María; Medicina y Cirugía Animal
Background: This study is focused on the measurement of trace elements (zinc, copper, cadmium, lead and selenium) in the saliva of pigs in order to study their levels on different porcine pathological conditions in the field. The experiment involved 15 pigs without clinical signs of disease and 42 diseased pigs (suffering from lameness, rectal prolapse, fatigue or growth rate retardation). Individual saliva samples were collected, allowing the pigs to chew a sponge each for trace element quantifications through atomic absorption spectrometry (AAS). Since this is the first report on the measurements of trace elements in porcine saliva, a routine analytical validation study was performed for the quantification of all the studied elements. Moreover, the acute phase proteins C-reactive protein (CRP) and haptoblobin (Hp), the total antioxidant capacity (TAC) and adenosine deaminase (ADA) were quantified in the saliva samples for the animal’s health status assessment. Results: Modifications in the levels of acute phase proteins or ADA were only recorded in animals with lameness and rectal prolapse and those with fatigue respectively. Moreover, TAC level changes were observed in pigs with growth-rate retardation. However, alterations in the levels of two or more trace elements were reported for all the different groups of diseased pigs with evident variations within pathologies. Conclusions: The salivary quantification of trace elements could be considered as a complementary tool to acute phase proteins, TAC and ADA determinations for disease detection and differentiation in the pig and should be explored in greater depth.
Embargo
Analytical validation of an automated assay for the measurement of adenosine deaminase (ADA) and its isoenzymes in saliva and a pilot evaluation of their changes in patients with SARS-CoV-2 infection
(2021-04-28) Franco Martínez, Lorena; Tecles, Fernando; Torres Cantero, Alberto; Bernal, Enrique; San Lázaro, Indra; Alcaraz, María José; Vicente Romero, Rosario; Lamy, Elsa; Sánchez Resalt, Cristina; Rubio, Camila P.; Tvarijonaviciute, Asta; Martínez Subiela, Silvia; Cerón, José J.; Medicina y Cirugía Animal
Objectives: The aim of the present study was to validate a commercially available automated assay for the measurement of total adenosine deaminase (tADA) and its isoenzymes (ADA1 and ADA2) in saliva in a fast and accurate way, and evaluate the possible changes of these analytes in individuals with SARS-CoV-2 infection. Methods: The validation, in addition to the evaluation of precision and accuracy, included the analysis of the effects of the main procedures that are currently being used for SARS-CoV-2 inactivation in saliva and a pilot study to evaluate the possible changes in salivary tADA and isoenzymes in individuals infected with SARS-CoV-2. Results: The automated assay proved to be accurate and precise, with intra- and inter-assay coefficients of variation below 8.2%, linearity under dilution linear regression with R2 close to 1, and recovery percentage between 80 and 120% in all cases. This assay was affected when the sample is treated with heat or SDS for virus inactivation but tolerated Triton X-100 and NP-40. Individuals with SARS-CoV-2 infection (n=71) and who recovered from infection (n=11) had higher mean values of activity of tADA and its isoenzymes than healthy individuals (n=35). Conclusions: tADA and its isoenzymes ADA1 and ADA2 can be measured accurately and precisely in saliva samples in a rapid, economical, and reproducible way and can be analyzed after chemical inactivation with Triton X-100 and NP-40. Besides, the changes observed in tADA and isoenzymes in individuals with COVID-19 open the possibility of their potential use as non-invasive biomarkers in this disease.
Embargo
Application of a score for evaluation of pain, distress and discomfort in pigs with lameness and prolapses: correlation with saliva biomarkers and severity of the disease
(2019-10) Contreras Aguilar, María Dolores; Escribano Tortosa, Damián; Martínez Miró, Silvia; López Arjona, Marina; Peres Rubio, Camila; Martínez Subiela, Silvia; Cerón Madrigal, José Joaquín; Tecles Vicente, Fernando; Medicina y Cirugía Animal
A score system was used to evaluate pain, distress and discomfort in healthy pigs and pigs with two different diseases: lameness and rectal prolapse. In addition, correlations between the results of this score and a panel of salivary biomarkers and severity of disease were studied. This panel included biomarkers of stress (cortisol, salivary alpha-amylase (sAA), total esterase activity (TEA), butyrylcholinesterase (BChE) and lipase (Lip)), im- munity (adenosine deaminase isozymes 1 (ADA1) and 2 (ADA2)) and oxidative status (uric acid (UA), Trolox equivalent antioxidant capacity (TEAC), cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of saliva (FRAS), advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2)). Based on their score, diseased animals were subdivided in those without and with evident pain. Lame pigs and prolapsed pigs with pain showed higher salivary levels of cortisol, sAA, TEA, BChE, ADA1 and ADA2 compared with the healthy pigs. In addition, the prolapsed pigs with pain showed higher levels of FRAS, AOPP and H2O2 compared with the healthy animals. Salivary cortisol, TEA, BChE, ADA isozymes 1 and 2, FRAS and AOPP correlated with the pain score. This five-point pain score system can be easily applied to lame and prolapsed pigs, and salivary bio- markers could be used as an additional tool for pain assessment in those pigs.
Open Access
Automated Spectrophotometric Assays for the Measurement of Ammonia and Bicarbonate in Saliva of Horses: Analytical Validation and Changes in Equine Gastric Ulcer Syndrome (EGUS)
(MDPI, 2024-02-28) Muñoz-Prieto, Alberto; Llamas-Amor, Eva; Contreras-Aguilar, María Dolores; Ayala de la Peña, Ignacio; Martín Cuervo, María; Cerón, José Joaquín; Hansen, Sanni; Medicina y Cirugía Animal
Ammonia (NH3) and bicarbonate (HCO3) have been related to gastric ulcers in humans. Ammonia is considered a possible cause of gastric ulcers, whereas bicarbonate has a protective function. The presence of ulcers in the stomach of horses is defined as Equine Gastric Ulcer Syndrome (EGUS), which is a frequent disease in this species, and it has been associated with changes in saliva composition, such as in analytes related to inflammation, immune system and oxidative stress. The objectives of this study were (1) to perform an analytical validation of two automated spectrophotometric assays, one for ammonia and one for bicarbonate, in the horses’ saliva and (2) to evaluate their possible variations with EGUS. Analytical validation of the automated assays for ammonia and bicarbonate in the saliva of horses showed that both assays were precise and accurate. In addition, significantly higher values of ammonia and lower values of bicarbonate were found in the saliva of horses with EGUS compared to healthy horses. It can be concluded that ammonia and bicarbonate can be measured in the saliva of horses and that ammonia increases and bicarbonate decreases in this sample type could be related to the presence of EGUS in this species.
Embargo
Biomarkers of oxidative stress in saliva of sheep: Analytical performance and changes after an experimentally induced stress
(Elsevier, 2019-04) Peres Rubio, Camila; Contreras-Aguilar, Maria Dolores; Quiles, A.; López-Arjona, Marina; Cerón, J.J.; Martínez-Subiela, Silvia; Hevia, M.L.; Escribano Tortosa, Damián; Tecles, Fernando; Medicina y Cirugía Animal
Oxidative stress can affect animal's health and the quality of its final products. The oxidative status can be evaluated by the measurement of both oxidant and antioxidant biomarkers. The use of saliva as a sample is preferable to blood, as individuals with limited training can collect it easily and non-invasively with minimal stress to the animal. The aim of this study was to perform an analytical validation of automated assays of the ferric reducing ability of plasma (FRAP), the cupric reducing antioxidant capacity (CUPRAC), the Trolox equivalent antioxidant capacity (TEAC) and uric acid as antioxidant biomarkers and of the advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2) as oxidant biomarkers in saliva samples of sheep, and to evaluate their possible changes after stress induced by shearing. All assays produced acceptable results in the analytical validation, from which it can be concluded that oxidative stress biomarkers such as FRAP, CUPRAC, TEAC, uric acid and AOPP and H2O2 can be measured in sheep saliva. In addition, acute stress due to shearing could produce an oxidative stress response in sheep and subsequently increase antioxidants in order to protect cells from damage.
Open Access
Changes in alpha-amylase activity, concentration and isoforms in pigs after an experimental acute stress model: an exploratory study
(2018-08-29) Contreras Aguilar, María Dolores; Martínez Subiela, Silvia; Martínez Miró, Silvia; Cerón Madrigal, José Joaquín; Tecles Vicente, Fernando; Medicina y Cirugía Animal; Department of Food and Animal Science, School of Veterinary Medicine, Universitat Autònoma de Barcelona
Background: Salivary alpha-amylase (sAA) is considered a non-invasive biomarker of acute stress that can be evaluated by changes in activity and concentration, and also by changes in its isoforms, although this last way of evaluation has never been used in veterinary medicine. This research evaluated the changes of sAA by three different ways in which sAA can be evaluated in an experimental acute stress model in six pigs based in a technique of temporarily restraining. These ways of evaluation were 1) activity by a spectrophotometric assay, 2) concentration by a fluorometric assay, and 3) isoforms of the enzyme by a Western blot. Results: Although salivary cortisol significantly increased due to the stimulus of stress and all the pigs manifested signs of stress by high-pitched vocalization, sAA activity showed an increase of different degree in the six pigs after the stress stimulus, while sAA concentration showed decreases in four of the six pigs. sAA activity did not correlate with sAA concentration or salivary cortisol, and a low correlation was observed between sAA concentration and salivary cortisol (r= 0.48, p = 0.003). The inter-individual variability was higher in sAA activity than in sAA concentration and salivary cortisol. Finally, three possible isoforms of sAA at 154-160 kDa, 65-66 kDa and 59-60 kDa were observed that showed different dynamics after the stress induction. Conclusions: Although this pilot study's results should be taken with caution due to the low sample size, it reveals a different behavior between sAA activity and concentration in pig after an acute stressful stimulus leading to evident external signs of stress by high-pitched vocalization, and opens a new field for the evaluation of possible selected isoforms of sAA as potential biomarkers of stress.
Open Access
Changes in Calprotectin (S100A8-A9) and Aldolase in the Saliva of Horses with Equine Gastric Ulcer Syndrome
(MDPI, ) Muñoz-Prieto, Alberto; Contreras-Aguilar, María Dolores; Cerón, José Joaquín; Ayala de la Peña, Ignacio; Martín-Cuervo, María; David Eckersall, Peter; Holm Henriksen, Ida-Marie; Tecles, Fernando; Hansen, Sanni; Medicina y Cirugía Animal
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.
Open Access
Changes in Oxidative Status Biomarkers in Saliva and Serum inthe Equine Gastric Ulcer Syndrome and Colic of IntestinalAetiology: A Pilot Study
(MDPI, 2022-03-07) Contreras-Aguilar, María Dolores; Peres Rubio, Camila; González-Arostegui, Luis Guillermo; Martín-Cuervo, María; Cerón, Jose J.; Ayala de la Peña, Ignacio; Holm Henriksen, Ida-Marie; Jacobsen, Stine; Hansen, Sanni; Medicina y Cirugía Animal
Changes in the oxidative status of the blood of horses suffering from gastric ulcers and colicof intestinal aetiology (CIE) have been reported. However, saliva can also be a source of biomarkersof oxidative status. Therefore, this study aims to validate automated assays for the measurement ofoxidative status biomarkers (ferric reducing ability of saliva/serum—FRAS/FRAP, cupric reducingantioxidant capacity—CUPRAC, the Trolox equivalent antioxidant capacity—TEAC, uric acid, andadvanced oxidation protein products—AOPP) in the saliva and serum of horses, to assess theirchanges in the different ulcer gastric diseases (squamous—ESGD and glandular—EGGD) and CIE,and to evaluate their relationship with serum amyloid A (SAA), adenosine deaminase (ADA), andthe systemic inflammatory response syndrome (SIRS) status. The assays showed a low imprecisionand good linearity with enough sensitivity in both fluids. In EGGD, higher levels of FRAS, uric acid,and AOPP in saliva were observed compared to the healthy group, correlating with the salivaryADA levels. Horses with CIE showed increases in uric acid concentrations in serum associated with heir SIRS status and outcome of the disease. In conclusion, analytes related to the oxidative statuscan be measured in the saliva and serum from horses by automated assays, and some of them canpotentially be assessed as biomarkers in horses with gastric ulcers and CIE
Embargo
Changes in oxytocin concentrations in saliva of pigs after a transport andduring lairage at slaughterhouse
(Elsevier, 2020-08-31) López Arjona, Marina; Escribano, Damián; Mateo, Sandra V.; Contreras Aguilar, María Dolores; Pérez Rubio, Camila; Tecles Vicente, Fernando; Cerón Madrigal, José Joaquín; Martínez Subiela, Silvia; Medicina y Cirugía Animal
Oxytocin is associated with reproductive physiology but also with welfare and positive emotions. In this study,oxytocin was measured in saliva samples of 45 pigs that were collected before being transported to theslaughterhouse, at the time of arrival and 4 h after arrival to the slaughterhouse. Two previously validatedassays, one that measures free oxytocin and other that measures oxytocin linked to proteins, were used. Inaddition, cortisol, salivary alpha-amylase (sAA), total esterase activity (TEA), butyrylcholinesterase (BChE) andlactate dehydrogenase (LDH), which are biomarkers associated with stress and pain in pigs, were measured. Theresults showed a decrease in free and protein-linked oxytocin concentrations at 4 h after transport comparedwith the time before transport, while cortisol, sAA, TEA, BChE and LDH showed an increase at 4 h after transportcompared with the time before transport. Based on these results it can be concluded that the transport andlairage at slaughterhouse in the conditions of this study produce a decrease in oxytocin in the saliva of pigs thatcould indicate a reduced emotional well-being
Open Access
Changes in Proteins in Saliva and Serum in Equine Gastric Ulcer Syndrome Using a Proteomic Approach
(MDPI, 2022-05-02) Muñoz-Prieto, Alberto; Contreras-Aguilar, Maria Dolores; Cerón, Jose Joaquín; Ayala de la Peña, Ignacio; Martin-Cuervo, María; González-Sánchez, Juan Carlos; Jacobsen, Stine; Kuleš, Josipa; Beletić, Anđelo; Rubić, Ivana; Mrljak, Vladimir; Tecles, Fernando; Hansen, Sanni; Medicina y Cirugía Animal
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 relatedto 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.
Open Access
Changes in Saliva Analytes Correlate with Horses’ Behavioural Reactions to An Acute Stressor: A Pilot Study
(2019-11-18) Contreras Aguilar, María Dolores; Henry, Séverine; Coste, Caroline; Tecles Vicente, Fernando; Escribano Tortosa, Damián; Cerón Madrigal, José Joaquín; Hausberger, Martine; Medicina y Cirugía Animal; Université de Rennes, Université de Normandie, CNRS, Laboratoire Ethologie et humaine - UMR CNRS 6552, Station Biologique
Acute stress induces an array of behavioural reactions in horses that vary between individuals. Attempts to relate behavioural patterns and physiological responses have not always given clear-cut results. Here, we measured the changes in a panel of salivary components: salivary alpha-amylase (sAA), lipase, total esterase (TEA), butyrylcholinesterase (BChE), adenosine deaminase (ADA), and cortisol, and their potential link with horses’ behaviours after acute stress. Saliva samples were collected in nine riding horses subjected to a test consisting of opening an umbrella. Saliva sampling was obtained at a basal time point in the stall (T1), in the test indoor arena (T2), at a time of stress (T3), and 30 min (T4) and 60 min (T5) later. The horses’ behaviour was recorded at T3 for 1 min. sAA, lipase, TEA, and BChE showed significant changes along time, increasing at T3 for BChE, and decreasing at T4 for sAA and BChE. Butyrylcholinesterase appeared to be the most reliable predictor of behavioural responses, as it correlated with the index of emotionality, of laterality, and the occurrence of alarm signals, while sAA decreased when horses expressed quieter behaviours. These first results bring promising lines for novel, more precise physiological markers of acute stress in horses that can bridge the gap between behaviour and physiology.
Open Access
Changes in saliva analytes in equine acute abdominal disease: a sialochemistry approach
(2019-06-06) Contreras Aguilar, María Dolores; Escribano Tortosa, Damián; Martínez Subiela, Silvia; Martín Cuervo, María; Lamy, Elsa; Tecles Vicente, Fernando; Cerón Madrigal, José Joaquín; Medicina y Cirugía Animal
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 < 0.001), urea (2.3 fold, P = 0.001), total bilirubin (2.6 fold, P < 0.001), total proteins (3.2 fold, P < 0.001), phosphorus (P, 4.5 fold, P < 0.001) and alpha-amylase (sAA, 8.5 fold, P < 0.001). Total proteins, P and sAA showed sensitivities higher than 70% at their optimal cut-off points and a specificity of 100% in differentiating between healthy horses and those with acute abdominal disease. Conclusions: A panel of 23 biochemical analytes can be measured in saliva of horses, where gGT, CK, urea, total bilirubin, total protein, P and sAA levels are raised in horses with acute abdominal disease.
Open Access
Changes in saliva biomarkers during a standardized increasing intensity ﬁeld exercise test in endurance horses
(Elsevier B.V., 2021) Contreras-Aguilar, María Dolores; Cerón, J.J.; Muñoz, A.; Ayala de la Peña, Ignacio; Medicina y Cirugía Animal
Salivary biomarkers could be useful to evaluate stress, ﬁtness level, and skeletal muscle damage associ-ated to exercise in horses in an easy and non-painful way. Therefore, this study aims to evaluate if cortisol in saliva (sCor), salivary alpha-amylase (sAMY) and butyrylcholinesterase (sBChE) and lactate (sLA) and creatine kinase (sCK) in saliva of horses can show changes during a standardized exercise test, and if they are related to heart rate variability (HRV) parameters related to sympathetic and parasympathetic tone, ﬁtness level or skeletal muscle damage. For this purpose, ten endurance horses were submitted to a stan-dardized exercise test in ﬁeld conditions. Saliva and blood were obtained at basal time (TB), after the seven bouts of velocity (T + 01 to T + 07), and 5, 15, 30, and 45 min later (T + 5, T + 15, T + 30, and T+ 45). Five endurance horses in resting condition (control group) were also enrolled. HRV and ﬁtness level parameters, and plasma CK as a marker of muscle damage were also evaluated. Salivaryalpha-amylase increased at T + 30 (P = 0.03), sBChE at T + 5 (P = 008), and sCK at T + 07 (P = 0.009) after the exercise test, with signiﬁcant differences between the exercise and control groups’ results. The sCor did not show sig-niﬁcant changes during the exercise test in the exercise group but higher concentration compared to the control horses (P < 0.001) were observed. sCor, sAMY, sBChE, and sCK showed a positive correlation (r val-ues between 0.47 and 0.64) with the sympathetic tone and a negative correlation (r values between 0.37 and 0.56) with the parasympathetic tone. In conclusion, sAMY, sBChE, and sCK showed signiﬁ-cant increases in ten endurance horses after an increasing intensity velocity exercise. Values of sCor, sAMY, sBChE, and sCK were associated with HRV, which is used to evaluate stress, and therefore, they could be potentially used to assess the exercise-related stress after a physical effort.
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Changes in salivary proteins can reflect beneficial physiological effects of ejaculation in the dog
(2021-01-28) Muñoz-Prieto, Alberto; Escribano, Damián; Horvatić, Anita; Contreras-Aguilar, María Dolores; Bernal, Luis; Rubić, Ivana; Cerón, Jose Joaquín; Dabrowski, Roman; Mrljak, Vladimir; Medicina y Cirugía Animal
The objective of this study was to study the changes in salivary proteins that occur in the dog after the ejaculation process. Saliva samples from eight dogs before and after induced ejaculation were analyzed by proteomic using Tandem Mass Tag (TMT) labeling and LC-MS/MS analysis. A total of 33 salivary proteins showed significant changes after the ejaculation process. The up-regulated proteins that showed changes of higher magnitude were mucin-7 (MUC-7), peroxiredoxin-4 (PRDX4) and galectin-3 (LEGALS3) whereas proteins such as alpha-1-acid glycoprotein (A1G1) and alpha-1B-glycoprotein (A1BG) were the most down-regulated. MUC-7 and PRDX4 expression in saliva after ejaculation could be associated with the protective “environment” created by the organism to exert pr 3o-fertility activities and antioxidants benefits in spermatozoa. Also LEGALS3 increment could be associated with an improvement of wellbeing and could contribute to a positive global effect in the body. Down-regulations of A1G1 and A1GB proteins found in saliva after ejaculation could be associated with a reduction in systemic inflammation. Overall it can be concluded that, changes in proteins in saliva that are produced after ejaculation can reflect a state of increase immune defenses, improvement of antioxidant status and low inflammation.