Browsing by Subject "Pigs"
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- PublicationOpen AccessA 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 AnimalSepsis 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
- PublicationOpen AccessArtificial insemination of all ejaculated sperm fractions accelerates embryo development and increases the uterine vascularity in the pig(Elsevier, 2024-04-15) Toledo Guardiola, Santa María; Luongo, Chiara; García Vázquez, Francisco A.; Soriano Úbeda, C.; Matás, C.; Párraga Ros, Ester; Seva Alcaraz, Juan; Anatomía y Anatomía Patológica ComparadasThe semen of boar is characterized by ejaculation in well-differentiated fractions with specific concentration, composition, and volume. The ‘sperm-rich fraction' (SRF), the most concentrated seminal fraction, is habitually collected in insemination centers to make artificial insemination (AI) doses. The absence of the other fractions in AI doses could alter the uterine reaction to AI and not trigger essential responses that could maximize fertility. Thus, there is an urge to ascertain the impact of different ejaculate fractions on the uterus after AI to optimize the semen doses. This work analyzed specific parameters related to fertility in pregnant artificially inseminated sows (n = 15) with ac-cumulative fractions of the semen of boars (n = 6): F1, composed of the sperm-rich fraction (SRF); F2, composed of F1 plus the intermediate fraction; F3, composed of F2 plus the post-SRF. Non-inseminated sows (n = 5) were included as control (C). The different types of seminal dose did not affect the number of ovulated follicles (CL; corpora lutea, p > 0.05) but did affect the embryo development (p < 0.05). The proportion of embryos in morula stages was significantly higher in AI-F1 sows (84.4%, p < 0.05). Morulas and blastocysts were balanced in AI-F2 or AI-F3 (p > 0.05). Independently of the type of seminal dose (F1, F2, or F3), we observed by immunohistochemistry that AI significantly increased uterine vascularization, although with some anatomical differences. The cranial region of the uterine horns was significantly more vascularized in AI-F1 or AI-F2 sows (26.7 ± 2.3 and 28.6 ± 2.0%, respectively), and AI-F3 showed significantly less vascularization at that point (17.8 ± 1.6%, p < 0.05). To summarize, the synergistic effect of all ejaculate fractions accelerates embryo development, at least during the preimplantation period, and increases the uterine reaction to AI in certain parts of the uterus.
- PublicationEmbargoC-reactive protein measurements in meat juice of pigs(Elsevier, 2008-04-15) Gutiérrez, A.M.; Martínez Subiela, Silvia; Montes, A.; Parra Muñoz, María Dolores; Cerón Madrigal, José Joaquín; Ciencias SociosanitariasA time-resolved immunofluorometric assay was evaluated for measurement of C-reactive protein in meat juice from diaphragmatic muscle collected from slaughtered pigs. Analytical and clinical validation of the method was performed by using meat juice samples, obtained by freezing and thawing muscle pieces. The intra- and inter-assay coefficients of variation ranged from 2.2–5.8% to 7.9–14.3%, respectively. The limit of detection was 0.00038 mg/ml. The method measured the CRP concentrations in a linear manner with a good accuracy (r = 0.99). CRP concentrations in serum were highly correlated with those in diaphragmatic meat juice (r = 0.90; p < 0.001). CRP concentrations were significantly higher in clinically affected pigs compared to non-diseased pigs. The assay described here provides a sensitive method for measuring CRP concentrations in meat juice, which can represent a suitable alternative to serum or blood samples and simplifies the process of sampling collection at slaughter.
- PublicationOpen AccessChanges in saliva proteins in two conditions of compromised welfare in pigs: T an experimental induced stress by nose snaring and lameness(Elsevier, 2019-06-19) Escribano Tortosa, Damián; Horvatić, Anita,; Contreras Aguilar, María Dolores; Guillemin, Nicolas,; Cerón Madrigal, José Joaquín; Tecles Vicente, Fernando; Martínez Miró, Silvia; Eckersall, Peter David; Manteca, Xavier; Mrljak, Vladimir; Producción AnimalThe aim of this study was to identify biological pathways and proteins differentially expressed in saliva of pigs in two conditions of compromised welfare: an acute stress consisting of restraint with a nose snare and in pigs with lameness which is a highly frequent problem in the swine industry. For this purpose, high-resolution quantitative proteomics based on Tandem Mass Tags labelling was used. Four proteins showed significant differences in the conditions of compromised welfare, namely cornulin, the heat shock protein 27 and lactate dehydrogenase (LDH), that showed significant increases, whereas immunoglobulin J chain showed a significant decrease. LDH, which was the protein that showed the highest differences, was selected for validation and clinical evaluation as a diagnostic biomarker. Significant changes in this protein were observed between pigs restrained with a nose snare and pigs with lameness compared with healthy pigs when measured with available commercial assays in a larger population of pigs. In conclusion, this study reports that in situations of compromised welfare on farm, such as acute stress and lameness in pigs, there are changes in proteins and metabolic pathways in saliva, and describes a series of proteins that could potentially be used as biomarkers for both short term acute stress and longer term chronic stress of lameness. These biomarkers would have the advantage of being measured in saliva by a noninvasive and not stressful collection sampling procedure.
- PublicationEmbargoEffect of repeated administration of lipopolysaccharide on inflammatory and stress markers in saliva of growing pigs(Elsevier, 2014-04-16) Campos, Paulo H.R.F.; Gutiérrez Montes, Ana María; Le Floc’h, Nathalie; Cerón Madrigal, José Joaquín; Merlot, Elodie; Escribano Tortosa, Damián; Producción AnimalAlthough saliva could be considered to be an ideal biological sample for evaluation of biomarkers relat- ing to stress and inflammatory responses in pigs, little is known about how these might be influenced by the presence of endotoxaemia. In the present study, the response to repeated administrations of li- popolysaccharide (LPS) was investigated, using a panel of salivary stress markers such as chromogranin A (CgA) and cortisol, as well as inflammatory/immune markers such as haptoglobin (Hp), C-reactive protein (CRP) and immunoglobulin A (IgA). Sixteen growing pigs were adapted to experimental conditions for 3 weeks, after which, 10 of the pigs were selected to receive three doses of LPS at 48 h intervals. Saliva samples were taken from all pigs prior to any LPS administration (baseline) and at time points corresponding to 3 h after each injection of LPS (T1, T2 and T3). Results showed that repeated administration of LPS induced significant elevation of salivary markers of hypothalamic-pituitary-adrenal (cortisol) and immune (Hp, CRP and IgA) activity compared to base- line levels (P < 0.05). However, rectal temperature, CRP and cortisol data suggested that the amplitude of the inflammatory response decreased with successive LPS administrations. Thus, measurement of sal- ivary biomarkers could be a practical tool for evaluating the inflammatory response to endotoxaemia in pigs. In the case of chronic inflammatory states, salivary Hp and IgA might be more sensitive markers than CRP or cortisol.
- PublicationOpen AccessGlycaemic response in pigs derived from assisted reproductive technologies: a long-term follow-up study(Society for Study of Reproduction (SSR), 2024) Romar, Raquel; Heras, Sonia; Paris-Oller, Evelyne; Soriano-Ubeda, Cristina; Gadea, Joaquin; Coy, Pilar; Canovas, Sebastian; Romero Aguirregomezcorta, Jon; FisiologíaEmerging evidence suggests that assisted reproductive technologies (ART) may predispose individuals to an increased risk of metabolic disorders including diabetes. Previous studies have reported a different glycaemic response in growing pigs derived from ART, but it is unknown whether these differences persist into adulthood and old age. The aim of the present work was to gain insight into the evolution of glycaemic response of ART-derived pigs across their life. Pigs born through artificial insemination (AI group) and transfer of in vitro-produced embryos (IVP group) underwent a glucose challenge at young age (45 days), adulthood (365 days), and old age (1250 days). The number of animals, respectively for AI and IVP groups, were 16 and 29 (young), 13 and 21 (adult), and 9 and 13 (old). Oral glucose tolerance test (OGTT; 1.75g/kg body weight) was performed after overnight fasting and 1h water withdrawn. Blood samples were obtained from auricular vein prior (T=0 min) and 15, 30, 45, 60, 90, 120, 150 min following glucose intake. Glycemia was immediately determined by glucometer test strips (GlucoMenLX Plus). The area under the curve analysis was used to identify differences (p < 0.05) between age and experimental groups. Minimum and maximum glycemia recorded during the three OGTT tests were 22.0 and 319.0 mg/dl respectively. Both the origin of the animals (AI vs. IVP) and the age (young-adult-old) significantly affected the glycaemic response (p < 0.05). At a similar age, there were no differences in fasting glucose between AI and IVP groups (young: 81.70±4.67 and 90.25±8.27 mg/dl; adult: 91.13±7.41 and 87.70±7.14 mg/dl; old: 65.12±3.50 and 59.93±4.12 mg/dl) being all data withing the physiological range for pigs (60.00-100.00 mg/dl). Mean glycaemia during OGTT was 25-30 mg/dl higher in young and adult animals compared to old pigs (p < 0.05).Evolution of glycemia curves showed a monophasic pattern during youth in bothgroups, as it is commonly observed in young pigs and we have previously described. Interestingly, the response pattern shifted into a biphasic curve in adult-AI pigs, returning to a monophasic response during old age, whereas in IVP animals the biphasic response appeared only during old age. The lack of studies in older pigs makes difficult to interpret the biphasic response which in other species is associated with a certain resistance to insulin. Moreover, differences between origin of animals were observed at young and old ages (p < 0.05). Therefore, young-IVP animals had higher AUC during the initial response to glucose challenge (15, 30 and 45 min) than young-AI pigs. However, old-IVP had lower AUC than old-AI pigs at the intermediate time of 60 min due to the biphasic glucose peak around this time. No differences between groups were observed during adulthood. In conclusion, naturally and artificially conceived pigs show different responses to glucose challenge across life. The clinical relevance and implications of such differences should be further studied.
- PublicationOpen AccessGrowth parameters and their hormonal regulation in pigs are differently affected by sex and embryo origin(2024) Heras, Sonia; Quintero-Moreno, Armando; Paris-Oller, Evelyne; Soriano-Ubeda, Cristina; Gadea, Joaquin; Romar, Raquel; Canovas, Sebastian; Coy, Pilar; FisiologíaThe global pig embryo trade has yet to emerge but replacing live animal transport with in vitro produced (IVP) embryos is a future objective. Understanding the potential long-term effects of in vitro protocols, especially in commercial traits like growth rates, is crucial. Our study aimed to compare growth parameters (body weight and length) and growth-related hormones (IGF-1 and T4) between IVP and artificially inseminated (AI) animals. Three experimental groups were defined, including 68 crossbred (Landrace x Large White) pigs that were allocated from a colony of animals produced in a previous study. IVP animals produced using BSA (C-IVP; 8 males, 16 females) or reproductive fluids (RF-IVP; 10 males, 7 females) as supplement for in vitro culture, were compared to AI animals (AI; 9 males, 18 females). Paternally, all animals were sired by the same boar, while maternally, IVP animals were originated from ovaries obtained at a local abattoir. Growth was measured at birth, 3, 9, 15, every 15 days until 180, 365, 700, 900, 1100, 1250, 1450, 1650 and 1800 days of age. Blood was collected at 90, 180, 365, 700 and 900 days of age. Plasma hormone concentrations were determined using a solid phase, enzymelabeled competitive chemiluminescent immunoassay. Data were analyzed using a linear mixed-effects model, with group, sex and age as fixed effects. Benjamini-Hochberg procedure was used for p-value correction and differences were considered significant when P< 0.05. Weight exhibited an exponential increase up to 6 months of age followed by a linear rise from 12 months of age. Levels of T4 decreased with age, and IGF-1 levels also decreased with age, but only in females and C-IVP males. Males were longer and heavier than females throughout the study. Additionally, males exhibited higher IGF-1 levels than females, while the opposite was observed for T4. Only in males, T4 levels negatively correlated with weight, and no correlation was observed between T4 and body length. IGF-1 positively correlated with weight and length until 6 months of age. IVP animals were significantly heavier (≤d180: 10.6 ± 1.1 kg; ≥365: 35.9 ± 5.4 kg) and longer (≤d180: 8.5 ± 0.8 cm; ≥365: 10 ± 1.9 cm) than those originated from AI throughout the study. In addition, C-IVP animals were heavier (4.3 ± 1.5 kg) and longer (3.2± 1.1 cm) than RF-IVP but only up to 6 months of age. Finally, in males, T4 levels of AI were higher than those of their IVP counterparts (1 ± 0.2 μg/dL) and IGF-1 levels of RF-IVP were higher than those of AI (36.1 ± 11.5 ng/mL). In conclusion, the growth and growth-related hormones of males and females exhibited differences throughout the study. Specifically, males exhibited higher levels of IGF-1, while females exhibited higher levels of T4. While statistical differences were identified between experimental groups, further studies with larger sample sizes are required to determine the clinical sign ificance of these findings, if any.
- PublicationRestrictedOn the aggregated nature of chronic Sarcoptes scabiei infection in adult pigs(Elsevier, 2012-10-17) Ruiz de Ybáñez Carnero, María del Rocío; Martínez Carrasco-Pleite, Carlos; Balseiro, A.; Alonso de Vega, Francisco; Casais, Rosa; Prieto, M.; García-Marín, J.F.; Berriatua Fernández de Larrea, Eduardo; Goyena Salgado, Elena; Sanidad AnimalThe prevalence and body distribution of Sarcoptes scabiei and associated dermatitis was investigated in sows and boars from four herds with long standing mange. Macroscopic hyperkeratotic dermatitis (crusted mange) was present in 1–6% of herd sows. Mite estimated prevalence (95% CI) in ear scrapings was 11% (6–17%) including 100% (13/13) and 2% (3/134) in sows with and without crusted mange, respectively, and the later had very few mites compared to the former. S. scabiei body distribution and dermatitis were further investigated in 59–64 skin scrapings/sow taken post-mortem from four culled sows including two (sows 1 and 2) with and two (sows 3 and 4) without crusted mange. The proportion of skin samples with eggs, instars or adults was 59% in sow 1, 84% in sow 2, 0% in sow 3 and 3% in sow 4. S. scabiei distribution in sows 1 and 2 ranged from being present in all skin ear and head samples to absent in those from the inner side of the limbs and mammary glands. Crusted lesions were observed in the skin of the ears, neck and lower limbs and contained the largest mite populations. Histopathological analysis of skin samples identified mites, inflammatory cellular infiltrate (mainly lymphocytes, neutrophils and eosinophils) and hyperkeratosis, acanthosis and spongiosis in 78%, 54%, 20% and 25% of samples from sows 1, 2, 3 and 4, respectively, being lesion severity positively associated to mite presence. The study provides further evidence that in herds with long-standing exposure to S. scabiei, infection becomes highly overdispersed with large mite populations present only in a few pigs and in specific body areas. Although the reasons for mite aggregation have not been identified, it is important controlwise because treating or eliminating a few and easy to identify heavily infected adult pigs, should markedly decrease the herd’s parasite load and reduce the use of acaridal drugs.
- PublicationEmbargoSaliva chromogranin A in growing pigs: a study of circadian patterns during daytime and stability under different storage conditions(Elsevier, 2014-01-17) Escribano Tortosa, Damián; Gutiérrez Montes, Ana María; Fuentes Rubio, María; Cerón Madrigal, José Joaquín; Producción AnimalSalivary chromogranin A (CgA) is considered to be a biomarker of activation of the sympatho-adrenome- dullary system, and has recently been proposed as a useful indicator of the acute stress response in pigs. The aim of the present study was to determinate whether salivary CgA concentrations in healthy growing pigs exhibits any circadian pattern during the daytime, and to evaluate its stability under different stor- age conditions. A total of 80 pigs (40 in spring and another 40 in autumn) of two different ages and gen- ders were used. To establish the circadian pattern, saliva samples were collected at 07.00, 11.00, 15.00 and 19.00 h on two consecutive days. Pooled samples were used for the stability study and were mea- sured on the day of sampling and periodically for up to 360 days later. Samples were stored at 4 °C, 20 °C or 80 °C and the effect of repeated freezing and thawing was also evaluated. No circadian pattern was detected for salivary CgA in either season and there were no significant effects of gender or age. However, mean salivary CgA concentrations were significantly higher (P < 0.0001) in the pigs sampled in autumn, compared to those sampled in the spring. Short term storage at 4 °C is recom- mended for up to 2 days, whereas frozen samples can be stored for 1 year at 20 °C or 80 °C, without substantial reduction in CgA values. In addition, samples can be frozen and thawed up to seven times without significant loss of the biomarker.
- PublicationOpen AccessThe impact of in vitro embryo production on placental and umbilical cord vascularization is minimized by the addition of reproductive fluids(Elsevier, 2023-09-15) Álvarez Martín, Úrsula; Coy, Pilar; Romar, Raquel; Párraga Ros, Ester; Seva Alcaraz, Juan; Anatomía y Anatomía Patológica ComparadasAnimals born from in-vitro-produced (IVP) embryos show changes in the placenta and umbilical cord vascularization. This study compares the placental and umbilical vascular morphometry in pigs (n = 19) born through artificial insemination (AI group) or after transfer of IVP embryos cultured with (RF-IVP group) or without (C-IVP group) reproductive fluids. The relationship between vascular parameters and animal growth during the first year of life was also analyzed. Samples were collected at birth, fixed, paraffin-embedded, cut in sections, stained, and photographed for vascular and morphometric analysis with ImageJ® and Slide Viewer®. The average daily weight gain was individually scored from birth to the first year of life. No differences were found in placental vascular morphometry among groups, except for the vascular area of small vessels (arterioles, venules, and small vessels) that was higher in the C-IVP group. Regarding the umbilical cord, the values for perimeter (AI: 26.40 ± 3.93 mm; IVP: 30.51 ± 4.74 mm), diameter (AI: 8.35 ± 1.01 mm; IVP: 10.26 ± 1.85 mm), area (AI: 43.18 ± 12.87; IVP: 56.61 ± 14.89 mm2), and Wharton's jelly area (AI: 36.86 ± 12.04 mm2; IVP 48.88 ± 12.80 mm2) were higher in IVP-derived than AI-derived animals, whereas arterial and venous morphometric data were similar between groups. A correlation study showed that placental and umbilical cord vascular phenotypes affect the further growth of pigs. In conclusion, assisted reproductive technologies impact small caliber vessels in the placenta and morphometric parameters in the umbilical cord. The addition of reproductive fluids in IVP-embryo contributes to reduce the differences with in vivo-derived animals.