Person: Gutiérrez Cepeda, Luna
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First Name
Luna
Last Name
Gutiérrez Cepeda
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Medicina y Cirugía Animal
Area
Medicina y Cirugía Animal
Identifiers
6 results
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Item Simple and economic colloidal centrifugation protocols may be incorporated into the clinical equine sperm processing procedure(Animal Reproduction Science, 2011) Gutiérrez Cepeda, Luna; Fernández, A; Crespo Castejón, Francisco; Gosálvez Berenguer, Jaime; Serres Dalmau, María ConsolacionFor many years in human assisted-reproduction procedures there have been special protocols to prepare and improve sperm quality. Colloidal centrifugation (CC) is a useful technique that has been proved to enhance semen quality by selection of the best spermatozoa for different species. Its use is recommended to improve fertility of subfertile stallions but current CC protocols are clinically complicated in the equine sperm processing technique due to economic and technical difficulties. The aim of this study was to determine the optimal processing procedures to adapt the use of a CC product (EquiPure™) in the equine reproduction industry. A total of nineteen ejaculates were collected from 10 Purebred Spanish Horses (P.R.E horses) using a Missouri artificial vagina. Gel-free semen aliquots were analyzed prior to treatment (control). Semen was subjected to one of six CC protocols with EquiPure™ and centrifuged samples were statistically evaluated by ANOVA and Duncan tests (p<0.05) for sperm quality and recovery rate. We obtained higher values by colloidal centrifugation in LIN, STR and BCF variables and DNA fragmentation index trended to be lower in most of the CC protocols. The studied protocols were shown to be as efficient in improving equine sperm quality as the current commercial EquiPure™, with the added advantage of being much more economical and simple to use. According to these results it seems to be possible to incorporate single layer and or high colloidal centrifugation volume protocols what would make them simple, economic and clinically viable for the equine sperm processing procedure.- Project number: 382
Item Elaboración de material docente para la enseñanza de la exploración física del caballo(2022) Fores Jackson, Paloma; Gutiérrez Cepeda, Luna; Villaescusa Fernández, Alejandra; Villalba Orero, María; Ruiz de León Robledo, María de los Ángeles; Crespo Castejón, Francisco; Dominuez Gimbernat, MonicaEl objetivo principal del presente proyecto fue incorporar una serie de recursos educativos virtuales que facilitaran e implementaran la enseñanza de la exploración física del caballo. Item Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation(Animals (based), 2022) Bláquez Sarro, Juan Carlos; Gutiérrez Cepeda, Luna; Crespo Castejón, Francisco; Serres Dalmau, María ConsolacionThe Purebred Spanish Horse, according to our clinical experience, is characterized by having a high number of stallions that do not meet the international commercial recommendations forequine-sperm cryopreservation. This means that artificial insemination with frozen semen from these stallions is less widespread than in other breeds. In this study, we investigated if the incorporation ofsingle-layer colloidal centrifugation prior to cryopreservation in clinical conditions could increase the number of ejaculates of Purebred Spanish stallions suitable for this processing, observing the influence of centrifugation and freezing extender protocol on post-thawed sperm motility. Using colloidal centrifugation, the percentage of ejaculates available to be frozen was increased from 35% (6/17) to71% (12/17), doubling the number of samples that could have been subjected to cryopreservation. We only found significant differences in linearity (LIN) and lateral head displacement (ALH) after5 min of incubation at 37 ◦C between colloidal and simple centrifugation processing techniques. No significant differences were found between the two different colloidal protocols in any of the variables considered. Colloidal centrifugation allowed us to obtain, from worse fresh-quality ejaculates, thawed sperm doses with similar quality to that of good-quality ejaculates. BotuCrio® produced, in general higher motility parameters and its characteristics than the other extenders analyzed, with significant differences found in comparison to Inra-Freeze® and Lac-Edta in both total (MOT) and progressive motility (PMOT) when using colloidal centrifugation and only in PMOT when applying simple centrifugation. Colloidal centrifugation optimized the efficiency of cryopreservation, as it allowed usto increase the number of ejaculates of Purebred Spanish Horses suitable to be frozen. Including these semen processing techniques in the freeze test could help to optimize equine-sperm cryopreservation protocols, especially when dealing with individuals or breeds for which initially low sperm quality prevents or limits their inclusion in sperm cryopreservation programsItem Strategies to Reduce the Use of Antibiotics in Fresh and Chilled Equine Semen(Animals, 2024) Zabala Argüelles, Sonsoles Mercedes; Serres Dalmau, María Consolacion; Montero Serra, Natalia; Crespo Castejón, Francisco; Lorenzo González, Pedro Luis; Pérez-Aguilera, Verónica; Galán López, Carmen; Domínguez Gimbernat, Mónica; Oliet Palá, Agustín; Moreno, Santiago; González Zorn, Bruno; Gutiérrez Cepeda, LunaThe study assessed the impact of four equine semen processing techniques on sperm quality and microbial load immediately post-processing and after 48 h of refrigeration. The aim was to explore the potential reduction of prophylactic antibiotic usage in semen extenders. Semen from ten adult stallions was collected and processed under a strict hygiene protocol and divided into four aliquots: Simple Centrifugation with antibiotics (SC+), Simple Centrifugation (SC−), Single-Layer Colloidal Centrifugation (CC−), and Filtration (with SpermFilter®) (F−), all in extenders without antibiotics. Sperm motility, viability, and microbial load on three culture media were assessed. No significant differences were observed in the main in the sperm quality parameters among the four protocols post-processing and at 48 h (p < 0.05 or p < 0.1). Microbial loads in Columbia 5% Sheep Blood Agar and Schaedler vitamin K1 5% Sheep Blood Agar mediums were significantly higher (p < 0.10) for raw semen than for CS+, CC−, and F− post-processing. For Sabouraud Dextrose Agar medium, the microbial load was significantly higher (p < 0.10) in raw semen compared to CS+ and F−. No significant differences (p < 0.10) were found in 48 h chilled samples. Regardless of antibiotic presence, the evaluated processing methods, when combined with rigorous hygiene measures, maintained semen quality and reduced microbial load to the same extent as a traditional protocol using antibiotics.Item Colloidal centrifugation of stallion semen results in a reduced rate of sperm DNA fragmentation(Reproduction Domestic Animals, 2013) Gosálvez Berenguer, Jaime; Crespo Castejón, Francisco; Gutiérrez Cepeda, Luna; Serres Dalmau, María Consolacion; Jonhnston, StephenStallion spermatozoa recovered and examined immediatelyafter colloidal centrifugation resulted in a higher straight-linevelocity (VSL) than sperm processed using direct conven-tional centrifugation (p = 0.000), but there was nodifferences in the progressive motility or sperm DNAfragmentation (SDF) as determined by the sperm chromatindispersion assay. However, when centrifuged spermatozoawere incubated at 37°C for 24 h to determine the rate of SDF(r-SDF), a lower r-SDF (p = 0.0011) was observed in thosesperm recovered after colloidal separation (0.5 ± 0.1%⁄h)compared to direct (1.2 ± 0.4%⁄h) or no centrifugation (r-SDF = 1.2 ± 0.3%⁄h). These results confirm that colloidalseparation of stallion spermatozoa results in prolonged spermDNA longevity, but these differences were only apparentfollowing a period of incubation and dynamic assessment.Consequently, we strongly recommend the use of thedynamic form of the SDF assay for evaluating centrifugationand⁄or otherex vivoprocedures, as a single basal assessmentof SDF may inadvertently result in a false-positive evaluationof DNA quality.Item The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA(Acta Veterinaria Scandinávica, 2012) Gutiérrez Cepeda, Luna; Fernández, Alvaro; Crespo Castejón, Francisco; Ramírez, Miguel Ángel; Gosálvez Berenguer, Jaime; Serres Dalmau, María ConsolacionVariability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method. Different extenders and processing techniques (such us colloidal centrifugation) are used in order to optimize post-thaw sperm quality. Sperm chromatin integrity analysis is an effective tool for assessing such quality. The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols (prior to cryopreservation) in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times (i.e., their DNA fragmentation dynamics). Results Post-thaw DNA fragmentation levels in semen samples subjected to either of the colloidal centrifugation protocols were significantly lower (p<0.05) immediately after thawing and after 4 h of incubation at 37°C compared to samples that underwent standard (control) centrifugation. The use of InraFreeze® extender was associated with significantly less DNA fragmentation than the use of Botu-Crio® extender at 6 h of incubation, and than the use of either Botu-Crio® or Gent® extender at 24 h of incubation (p<0.05). Conclusions These results suggest that single layer colloidal centrifugation performed with extended or raw semen prior to cryopreservation reduces DNA fragmentation during the first four hours after thawing. Further studies are needed to determine the influence of freezing extenders on equine sperm DNA fragmentation dynamics.