Prácticas virtuales de fisiología

El presente proyecto de innovación educativa ha servido para generar material audiovisual compatible con dispositivos electrónicos para el aprendizaje de fisiología de una manera sencilla y accesible. Es la continuación de un proyecto del curso 2020-21, en el que se generó un material con las prácticas que se realizan con un programa de registros fisiológicos. En esta ocasión, se procedió a la grabación de los procedimientos experimentales de las asignaturas de Fisiología del Grado en Veterinaria y del Grado en CyTA y se han incluido en un formato presentación, en el que se exponen los conocimientos básicos de la práctica, los objetivos, el material y métodos, el procedimiento experimental en vídeo con explicaciones y finalmente la recogida de datos y la interpretación de los resultados. Así, se reduce el número de animales a utilizar en las prácticas porque no es necesario volver a hacer el procedimiento en el animal y el estudiante puede aprender a su ritmo y visualizar la presentación tantas veces como necesite para su aprendizaje, antes, durante y después de la práctica. Se ha contado con un amplio equipo en el que han participado PDI del departamento de Fisiología y de Producción Animal, PAS y estudiantes de grado y posgrado, de modo que la experiencia de todos ellos ha aportado un enfoque multidisciplinar, muy adecuado para la viabilidad del proyecto. El producto generado es un material duradero en el tiempo, que seguirá poniéndose a disposición de los estudiantes en cursos venideros

Çédille, revista de estudios franceses

Presentació

Superoxide dismutase mimics improves semen quality during chilled preservation of rabbit spermatozoa

In rabbit farms, artificial insemination is usually undertaken using semen preserved around 18 ºC. However, increased reactive oxygen species levels during preservation produce sperm dysfunction. Our aim was to evaluate the effect of adding superoxide dismutase (SOD) antioxidant mimics (Tempo and TempoL) to the extenders of rabbit semen on quality parameters, total lipid peroxidation (LPO) and SOD level. Ejaculates from 12 sexually mature males were diluted with an extender containing 0 (control), 0.5, 1 or 2 mM of Tempo or TempoL, respectively. Semen samples were cooled till 18 ºC and stored for 0, 6 or 24 h. Sperm motility, velocity parameters, viability, plasma membrane integrity, SOD activity and level of thiobarbituric acid reactive substances (TBARS) were determined. Results showed that, in general, SOD antioxidant mimics significantly improved sperm motility compared to control after 6 and 24 h storage. Moreover, at 24 h, Tempo treatments showed motility rates higher than 80% whilst the control group showed the lowest motility rate among all treatments (58.5%, P < 0.05). Sperm viability showed no significant differences between treatments at 6 h and 24 h of storage. At 24 h, most SOD antioxidant mimics treatments significantly improved both curvilinear and mean velocity parameters, but only Tempo treatments improved linear velocity parameter. Also, SOD mimics protected sperm cells decreasing TBARS concentration at 6 and 24 h compared to 0 h. In conclusion, the addition of SOD antioxidant mimics during conservation at 18 ºC and storage of semen until 24 h reduces lipid peroxidation and preserves rabbit semen quality

Effects of a lignin-rich fibre diet on productive, reproductive and endocrine parameters in nulliparous rabbit does

In rabbits, modifications in dietary composition may be associated with reproductive benefits. However, the influence of nutrition on ovarian function and embryo development is poorly known in this species. The goal of this work was to study the short-term effects of feeding high-fibre diets with different levels of lignin during the rearing period on productive, endocrine and reproductive parameters of nulliparous rabbit does. A total of 95 New Zealand × California 11-week-old nulliparous white rabbits were randomly allocated in two experimental groups fed ad libitum fibre-rich diets with a high lignin content (HL group: NDF 49.6% of DM, LAD 15.8% of DM; n = 48) or a standard lignin content (SL group: NDF 40.9% of DM, LAD 4.9% of DM; n = 47). All animals were artificially inseminated (AI) at 16 weeks of age. Daily feed intake and live body weight (LBW) were recorded during the rearing period. Conception rate and prolificacy were also determined. In addition, in ten animals of each group, body composition, serum estradiol 17β (E2), progesterone (P4) and leptin levels, as well as reproductive parameters including ovary weight, follicular population, ovulation rate, in vitro oocyte maturation (IVM), blastocyst recovery rate and in vitro embryo development were studied at AI time. Animals fed the HL diet had a significantly higher feed intake until the first AI (P < 0.003) and during the first pregnancy (P < 0.03). At 16 weeks of age, animals fed the HL diet had a lower content of lipids (P < 0.05) and crude energy (P < 0.05) than those fed the SL diet, but average LBW of does, ovary weight, and ovulation rate per doe were similar between HL and SL treatments. No significant differences were found in P4 levels. However, the HL group had elevated E2 serum levels (P < 0.003) compared to those fed with the SL diet, whereas antral follicular population was similar between groups. Both nuclear and cytoplasmic IVM measured as metaphase II and cortical granule migration rates were also similar between both groups. In addition, no differences in blastocyst recovery rate or in prolificacy were found. However, mean serum leptin levels (P < 0.05), in vitro embryo development (P < 0.03), and fertility rate (P < 0.02) were higher in the SL group compared to the HL group. In conclusion, the HL diet enhanced feed intake of does, but reduced serum leptin levels at AI time, in vitro embryo survival and conception rate.Ministerio de Educación y CienciaUniversidad Complutense de MadridDepto. de Producción AnimalSección Deptal. de Fisiología (Veterinaria)Fac. de VeterinariaTRUEpu

Influence of metabolic status on oocyte quality and follicular characteristics at different postpartum periods in primiparous rabbit does

Low reproductive performance of high-yield primiparous animals is closely associated with the metabolic stress caused by a simultaneous gestation and lactation. The aim of this work was (1) to analyze body composition and metabolic environment at three time points along lactation (at parturition time; in the lactation period [Day 11 postpartum]; and in the postweaning period [Day 32 postpartum]) of primiparous rabbit does (Oryctolagus cuniculus) and (2) to investigate the ovarian status at insemination time and the possible link with metabolic environment and with their reproductive performance. To this end, does were either submitted to a semi-intensive reproductive rhythm (Group S, inseminated on Day 11 postpartum) or an extensive rhythm (Group E, inseminated on Day 32 postpartum). Body energy (P < 0.05) and protein content (P < 0.001) as well as serum leptin (P < 0.05) and protein concentrations (P < 0.001) increased significantly along the postpartum period. At parturition, body lipid content was significantly lower and serum nonesterified fatty acids concentrations were significantly higher than that on Days 11 postpartum and 32 postpartum. Concerning ovarian status at insemination time, no significant differences were found in mean follicular stages, serum estradiol, progesterone, and prolactin (PRL) concentrations or in prolactin receptor (PRL-R) immunostaining. However, follicles in Group S showed a significantly higher apoptosis index than that of Group E (P < 0.001). The nuclear and cytoplasmic oocyte maturation rates of Group S were also significantly lower than that in Group E. In addition, conception rate and prolificacy were improved in Group E (P < 0.001 and P < 0.05, respectively). In conclusion, in the early postpartum period, metabolic status seems to impact negatively on ovarian follicle and oocyte quality leading to a poor reproductive outcome in primiparous rabbit does.Sección Deptal. de Fisiología (Veterinaria)Fac. de VeterinariaTRUEpu

Indicadores metabólicos y de estrés tras el ajuste de consumo en conejas primíparas = Metabolic and stress indicators after intake adjustment in primiparous pregnant rabbit does

El objetivo principal de este trabajo fue determinar si aplicando un ajuste en el consumo de alimento durante la gestación en conejas primíparas, se ven afectados indicadores metabólicos y de estrés. Para ello, se utilizaron conejas primíparas, inseminadas después del destete que se dividieron en 4 grupos: Control (consumo od Iibitum toda la gestación, n=30), y restringidas al 60% de su consumo voluntario (1 OSg/d) la primera semana (RO?; n=31), la segunda y la tercera semana (R721; n=31) y las 3 primeras semanas (R021; n=30) de la gestación. En la cuarta y última semana de gestación todas disponían de pienso od libitum. Se tomaron muestras de sangre el día de la inseminación y tras el diagnóstico de gestación, se muestrearon animales gestantes de los 4 grupos en 3 puntos de la gestación el día 14 (n=20), 21 (n= 20) y 28 (n=20). Tras el parto, las hembras se re-inseminaron el día 14 post-parto y se tomaron muestras en otros 3 puntos de la lactación: día? (n=13), día 14 (n=13) y día 30 (n=41). En estas muestras se analizaron mediante enzimoinmunoanálisis: tri-iodotironina y tiroxina libres, y corticosterona. El ajuste del consumo de alimento durante la gestación aplicado en conejas primíparas afectó levemente a la homeostasis energética de los animales pero su estado de bienestar no se vio comprometido

Indicadores metabólicos y de estrés tras el ajuste de consumo en conejas primíparas = Metabolic and stress indicators after intake adjustment in primiparous pregnant rabbit does

El objetivo principal de este trabajo fue determinar si aplicando un ajuste en el consumo de alimento durante la gestación en conejas primíparas, se ven afectados indicadores metabólicos y de estrés. Para ello, se utilizaron conejas primíparas, inseminadas después del destete que se dividieron en 4 grupos: Control (consumo od Iibitum toda la gestación, n=30), y restringidas al 60% de su consumo voluntario (1 OSg/d) la primera semana (RO?; n=31), la segunda y la tercera semana (R721; n=31) y las 3 primeras semanas (R021; n=30) de la gestación. En la cuarta y última semana de gestación todas disponían de pienso od libitum. Se tomaron muestras de sangre el día de la inseminación y tras el diagnóstico de gestación, se muestrearon animales gestantes de los 4 grupos en 3 puntos de la gestación el día 14 (n=20), 21 (n= 20) y 28 (n=20). Tras el parto, las hembras se re-inseminaron el día 14 post-parto y se tomaron muestras en otros 3 puntos de la lactación: día? (n=13), día 14 (n=13) y día 30 (n=41). En estas muestras se analizaron mediante enzimoinmunoanálisis: tri-iodotironina y tiroxina libres, y corticosterona. El ajuste del consumo de alimento durante la gestación aplicado en conejas primíparas afectó levemente a la homeostasis energética de los animales pero su estado de bienestar no se vio comprometido

Recombinant rabbit beta nerve growth factor production and its biological effects on sperm and ovulation in rabbits

In some induced-ovulating species, beta nerve growth factor (β-NGF) has important roles in ovulation, though data for rabbits are still inconclusive. In this study we first synthesized functional recombinant β-NGF from rabbit tissue (rrβ-NGF) to address the following objectives: 1) to compare rabbit β-NGF amino acid sequence with those of other induced- or spontaneous-ovulating species; 2) to assess the effects of rrβ-NGF on rabbit sperm viability and motility, and 3) to examine the in vivo ovulation inducing effect of rrβ-NGF added to the seminal dose in rabbit does. The NGF gene in rabbit prostate tissue was sequenced by Rapid Amplification of cDNA Ends and annotated in GenBank (KX528686). Recombinant rβ-NGF was produced in CHO cells and purified by affinity chromatography. Once confirmed by Western blotting and mass spectrometry (MALDI-TOF) that the amino acid sequence of the recombinant protein corresponded to β-NGF, its functionality was validated in PC12 cells in a successful dose-response study over 8 days. The amino acid sequence of prostate rabbit NGF differed to that of other species mainly in its receptor binding sites. In all the spontaneous ovulating species examined, compared with rabbit, alanine and proline residues, which interact with the high-affinity receptor, were replaced by a serine. In rabbits, asparagine and methionine were substituted by lysine at the low-affinity receptor binding site. In time- and dose-response experiments, the in vitro addition of rrβ-NGF to the ejaculate did not affect sperm viability whereas sperm motility parameters were enhanced by the addition of 1 μg/mL of the neuropeptide. Addition of this same concentration of rrβ-NGF to the seminal dose administered via the intravaginal route in does induced ovulation with a delayed LH peak, leading to a plasma progesterone increase, gestation and delivery. Our findings suggest that rrβ-NGF could be a useful option for biotechnological and reproduction assisted techniques in rabbits but further studies are needed.Ministerio de Economía y Competitividad de España [subvención AGL2015-65572-C2]Contrato Predoctoral UCM-Santander de ASRContrato de Empleo Joven de la Consejería de Educación, Juventud y Deporte de la Comunidad de Madrid y el Fondo Social Europeo de PADepto. de Producción AnimalFac. de VeterinariaTRUEpu

A diet supplemented with n-3 polyunsaturated fatty acids influences the metabolic and endocrine response of rabbit does and their offspring

The aim of the present study was to evaluate the productive, endocrine, and metabolic responses as well as oxidative stress of rabbit does and their offspring when fed a diet supplemented with n-3 PUFA during their first productive cycle. To this aim, a total of 105 rabbit does were fed ad libitum from d 60 to 172 of age 2 isoenergetic and isoproteic diets differing in fatty acid composition. The control diet (n = 52 does) contained 45.9 g/kg of n-3 of the total fatty acids and the enriched diet (n = 53 does) contained 149.2 g/kg of n-3 of the total fatty acids. Both experimental groups had similar feed intake during rearing, pregnancy, and lactation. The enrichment of diet had no effect on ultrasonographic assessment of does on d 9 and 16 of pregnancy, with an embryonic vesicle number and fetus and placenta size similar between groups (P > 0.05). Even though there were no major effects (P > 0.05) on fertility, duration of gestation, and number born alive and stillborn kits at parturition, live kits from enriched does were longer (71.6 ± 2.42 vs. 79.5 ± 2.13 mm; P 0.05). In enriched does, higher plasma leptin and estradiol concentrations than in control does (P < 0.05) were observed. In addition, enriched females also had lower total and high-density lipoprotein cholesterol (HDL-c) than control females during lactation (P < 0.05). Regarding offspring, the enrichment of diet with PUFA caused a hyperlipidemic status (greater values of plasma triglycerides, total cholesterol, and HDL-c; P < 0.05) at 1 d postpartum (dpp), compared with the control group, that disappeared at 32 dpp. Supplemented does before parturition and their offspring at 1 dpp had greater oxidative stress than those in the control group. In conclusion, an increase of n-3 PUFA concentration in the diet of rabbit does and, consequently, of their offspring during a productive cycle alters their lipid profile and the indicators of oxidative stress, without major endocrine modifications or improvements in the productive variables.Ministerio de Economía y Competitividad (España)Comunidad de MadridSección Deptal. de Fisiología (Veterinaria)Depto. de Producción AnimalFac. de VeterinariaTRUEpu