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PublicationOpen AccessCould Estrogen and Progesterone be used as biomarkers in swine saliva?(2024) Quintero-Moreno AA, Serrano-Albal M, Barcelo Campoy MD, Romar R, Gadea J.; Fisiología- PublicationOpen AccessLong term study of the blood plasma biochemical profile of cattle born by assisted reproductive technologies(Brazilian College of Animal Reproduction, 2024) Serrano-Albal, Maria; Romero-Aguirregomezcorta, Jon; Canovas, Sebastian; Heras, Sonia; Quintero-Moreno, Armando; Lopes, Jordana Sena; Soriano-Ubeda, Cristina; Gadea, Joaquin; Coy, Pilar; Romar, Raquel; Departamentos::Departamentos de la UMU::FisiologíaAssisted reproductive technology (ART) is key in cattle breeding as it enables selective mating and improves both reproductive efficiency and genetic improvement. However, there is still limited information on the long-term effects of these methods on the health and development of the offspring. This study aims to investigate the biochemical profile of cattle born through ART, from a previous study (Lopes et al., 2020), over an extended period to understand how these techniques may affect their health and metabolism. Male and female (not pregnant or lactating) cattle born after artificial insemination (AI, N= 7) were compared to those derived from in vitro produced embryos (IVP) using a standard protocol (C-IVP, N= 7), or using oviductal and uterine fluids in the process (RF-IVP, N= 4). Males and females were kept in two diferent open pens, but fed and managed under identical conditions their whole lives. Animals were studied every six months, from 1.5 years old until ≥4 years old, since their first year of life was evaluated by Lopes et al. (2022). Tail vein blood plasma was obtained by centrifugation (1000 G, 10 min) in lithium heparin tubes and stored at 80°C. The biochemical profile included total protein (TP), albumin (ALB), globulin (GLOB), creatinine (CREA), urea (URE), glucose (GLUC), cholesterol (CHOL), triglycerides (TRIG), amylase (AMIL), lipase (LIP), creatinine kinase (CK), alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TB). After serial dilutions, inter- and intraassay precision of the methods were lineal and below 15%. Data normal ly distributed was analysed using ANOVA and Tukey’s test; data not normally distributed was analysed using the Kruskal Wallis and Games Howell’s test (statistical significance p<0.05). The data is shown below as means by group over the years. The AI group had lower CREA and CHOL levels (1.43±0.05 mg/dL and 109.97±5.27 mg/dL, respectively) than C-IVP (1.75±0.07 mg/dL and 126.57±7.53 mg/dL, respectively) and RF-IVP (1.86±0.08 mg/dL and 140.01±7.19 mg/dL, respectively). The GGT levels were lower in RFIVP animals (13.00±1.30 UI/L) than both the C-IVP (17.55±1.10 UI/L) and AI groups (16.05±1.23 UI/L). Moreover, the levels of ALT were significantly lower in C-IVP (28.60±1.72 UI/L) than in the AI (34.35±1.81 UI/ L) and RF-IVP groups (33.40±2.24 UI/L). Depending on the age of the animal at the time of sample collection, significant differences were observed for most parameters. The parameters TP, GLOB, CREA, URE, AMIL, AST an increased in their concentration was observed as the animals aged, whilst ALB, GLUC, ALP decreased with age. However, all values can be considered within physiological ranges despite the significant v ariations. To conclude, the study showed differences in some biochemical parameters amongst the ART groups and with aging. However, despite the diferences all values were within physiological ranges. These findings are key to study the potential implications of this diferences on the animal’s health.
- 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; Romero-Aguirregomezcorta, Jon; Heras, Sonia; Paris-Oller, Evelyne; Soriano-Ubeda, Cristina; Gadea, Joaquin; Coy, Pilar; Canovas, Sebastian; Departamentos::Departamentos de la UMU::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 AccessPreliminary Study of Sex Hormone Levels Throughout the Estrous Cycle in the Saliva of 4.5 year old Sows Born by ART(2024) Serrano Albal, María; Quintero-Moreno, Armando; Romero-Aguirregomezcorta, Jon; Gadea, Joaquín; Romar, Raquel; Department of Physiology, Universidad de Murcia, International Excellence Campus for Higher Education and Research (Campus Mare Nostrum), Murcia, Spain Institute for Biomedical Research of Murcia (IMIB), Murcia, SpainEstradiol (17-beta-estradiol, E2) and progesterone (P4) play a key role in the regulation of the estrous cycle. Their levels fluctuate throughout the cycle, triggering processes that are essential for the expression of mating behaviour. The rising levels of E2 during the follicular phase triggers the initiation of behavioural and physiological changes associated with heat in sows, whilst P4 is essential for early embryo development and uterine preparation for implantation. The evaluation of sex hormones is typically performed on plasma or serum. However, daily blood collection puts stress on the animals and requires specialized staff, and the extraction may alter the results of some metabolites. Since the small size of these molecules allows them to penetrate the membranes, such as those of the salivary glands, using saliva to measure sex hormones could offer a non-invasive approach to facilitate the collection of sex hormone data. Yet, studies on sex hormones in pig saliva are very scarce, limiting its application in the field. Here, the correlation between the levels of E2 and P4 were evaluated in plasma and saliva, at three separate timepoints, in 22 Large White and Landrace crossbreed sows. These animals were born through different assisted reproductive technologies from a previous study (Paris-Oller et al., 2021), with their ages around 4.5 years old. Their exclusion from production lines allows them to have natural estrous cycles, be kept in an open enclosure, and fed under identical conditions. Furthermore, 12 sows of the same herd [6 born via artificial insemination (AI group) and 6 born after surgical transfer of in vitro-produced embryos (IVP group)] were used to measure the fluctuation of both E2 and P4 in saliva during the estrous cycle. Plasma was obtained through the centrifugation (1000 G, 10 min) of ophthalmic venous sinus blood collected in lithium heparin tubes. Saliva samples were collected using Salivette® tubes containing a polystyrene sponge previously chewed by the sows for 10 secs. These tubes were then centrifuged (1000 G, 5 min) and the samples stored (-80ºC) until hormone analysis. Both plasma and saliva samples were later analysed by quimioluminescence (Immulite). The correlation between the levels of E2 in saliva and plasma, analysed using a Pearson correlation coefficient, showed a statistically significant correlation (p < 0.05) with a moderate negative relationship (-0.311). The average level of plasma E2 was 25.78 pg/mL whilst the saliva E2 was 49.05 pg/mL. However, the correlation between P4 in saliva and plasma was not significant, with an average concentration of 15.23 ng/mL and 0.54 ng/mL in plasma and saliva, respectively. Analyses of the hormone levels in saliva showed no significant differences for both hormone levels between the AI group and IVP group, nor for P4 levels among the days of the estrous cycle. Nevertheless, significant differences were found in the levels of E2 per day of the estrous cycle (p < 0.001). Moreover, in addition to the peak expected towards the end of the follicular phase, another peak was observed around days 12-14 of the cycle. This peak is essential for the maternal recognition of pregnancy in pigs and has been identified in blood but has never been reported in saliva of nonpregnant animals.
- PublicationOpen AccessRepercusión de un programa adaptado de aula invertida sobre la motivación intrínseca, el clima de autonomía y el rendimiento académico en el contexto universitario(Ediciones Octaedro, 2024-06) Arias-Estero, José L.; Meroño García, Lourdes; Morales-Belando, María T.; Expresión Plástica, Músical y Dinámica