Person: Crespo Castejón, Francisco
Loading...
First Name
Francisco
Last Name
Crespo Castejón
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Medicina y Cirugía Animal
Area
Medicina y Cirugía Animal
Identifiers
6 results
Search Results
Now showing 1 - 6 of 6
Item Fertilizing capacity of vitrified stallion sperm assessed utilizing heterologous IVF after different semen warming procedures(Animal Rerpoduction Science, 2020) Consuegra González, Cesar; Crespo Castejón, Francisco; Dorado Martín, Jesús; María Díaz Jiménez; Sánchez Calabuig, María Jesús; Beltrán Breña, Paula; Pérez Cerezales, Serafín; Rizos, Dimitri; Hidalgo PrietoThe aim of this study was to evaluate the fertilizing capacity of frozen or vitrified stallion sperm after assessing different warming procedures. In Experiment 1, different warming procedures were compared after sperm vitrification: immersion in extender at 43 ◦C (C), or in a water bath at 37 ◦C/30 s (W37), 43 ◦C/10 s (W43) or 60 ◦C/5 s (W60). With the W60 treatment, there were greater values (P < 0.05) for VCL (83.93 ± 3.6 μm/s) and ALH (3.00 ± 0.2 μm) than freezing and with the C group, and greater values (P < 0.001) for PM (35.33 ± 2.5 %) than with the W43 treatment. In Experiment 2, the fertilizing capacity of vitrified and frozen sperm was assessed utilizing heterologous IVF procedures, using cattle oocytes. Vitrification resulted in greater values (P < 0.05) than freezing for the number of bound sperm (1.36 ± 0.3 and 0.69 ± 0.2, respectively). There were no differences between frozen or vitrified sperm in pronuclear formation (26 hours post-insemination - hpi; 14.08 ± 4.2 % and 22.78 ± 4.8 %, respectively) or cleavage rate (32.77 ± 4.3 % and 39.66 ± 4.6 %, respectively). In conclusion, vitrified stallion sperm warmed in a water bath at 60 ºC had the capacity to penetrate cattle oocytes, leading to pronuclear formation and hybrid embryo cleavage after heterologous IVF.Item Comparison of different mathematical models to assess seasonal variations in the longevity of DNA integrity of cooled-stored stallion sperm.(Andrología, 2020) Rizos, Dimitri; Hidalgo Prieto; Hidalgo Prieto, Manuel; Consuegra González, Cesar; Diaz Jiménez, María; Dorado Martín, Jesús; Crespo Castejón, FranciscoDynamic assessment of sperm DNA fragmentation (SDF) has shown to give fuller understanding of stallion semen quality; however, there have been limited attempts to use this parameter to investigate seasonal changes in productive functions. The aims of this study were to: (a) establish a reliable mathematical model to describe the longevity of cooled-stored sperm DNA integrity; (b) to examine the effect of seasonal variations on SDF. Ejaculates were cooled to 5°C, and SDF was analysed after 0, 6 and 24 hr of storage. The coefficient of determination (R2) was calculated after fine-tuning linear (LIN), exponential (EXP) and second order polynomial (POL) models. R2 was significantly higher (p < .001) for POL than for LIN and EXP. The rate of DNA degradation was calculated using the slopes of POL equations. After assessing the rate of change of the POL functions, significant differences between the acceleration of DNA fragmentation were found (p < .01) among seasons, being higher for winter and summer than spring and autumn. In conclusion, DNA analysis of stallion sperm fits better to a second order polynomial mathematical model, being spring the best season to collect and process cooled stallion semen in order to maintain the DNA integrity of the stallion sperm.Item Effect of single-layer centrifugation or washing on frozen-thawed donkey semen quality: Do they have the same effect regardless of the quality of the sample?(Theriogenology, 2015) Ortiz Jaraba, Isabel; Dorado Martín, Jesús; Morrell, Jane M; Crespo Castejón, Francisco; Gosálvez Berenguer, Jaime; Gálvez, MJ; Acha Valls, Daniel; Hidalgo Prieto, ManuelThe aims of this study were to determine the sperm quality of frozen-thawed donkey sperm samples after single-layer centrifugation (SLC) using Androcoll-E in comparison to sperm washing or no centrifugation and to determine if the effect on the sperm quality after SLC or sperm washing depends on the quality of the sample. Frozen-thawed sperm samples from Andalusian donkeys were divided into three aliquots, and they were processed using three different techniques after thawing: uncentrifuged diluted control (UDC), sperm washing (SW), and SLC. Afterward, sperm quality index was estimated by integrating all parameters (total and progressive sperm motility, membrane integrity, and DNA fragmentation) in a single value. The relationship between the sperm quality of thawed UDC samples and the effect on sperm parameters in SW and SLC-selected samples was assessed. Sperm quality index was significantly higher (P < 0.001) in SLC (0.8 ± 0.0) samples than that in UDC (0.6 ± 0.0) and SW (0.6 ± 0.0) samples, regardless of the sperm quality index after thawing of the sperm sample. In conclusion, SLC of frozen-thawed donkey spermatozoa using Androcoll-E-Small can be a suitable procedure for selecting frozen-thawed donkey sperm with better quality, in particular in those samples where an improvement in motility is needed.Item Stallion sperm selection prior to freezing using a modified colloid swim-up procedure without centrifugation(Animal Rerpoduction Science, 2017) Hidalgo Prieto, Manuel; Ortiz Jaraba, Isabel; Dorado Martín, Jesús; Morrel, Jane M; Gosálvez Berenguer, Jaime; Consuegra González, Cesar; Diaz Jimenez,María; Pereira Aguilar, Blasa; Crespo Castejón, FranciscoThe aims of this study were to: 1) develop a new method for stallion sperm selection using a modified swim-up procedure through a colloid and 2) evaluate its impact in good quality eja-culates from bad freezers in comparison to methods involving centrifugation such as single layer centrifugation and sperm washing. Ejaculates were processed before freezing using three dif-ferent procedures: sperm washing (SW), colloid single layer centrifugation (SLC) and a modified colloid swim-up (SU). After semen processing, sperm recovery rates were measured and sperm were frozen. Post-thaw sperm motility (assessed by computer-assisted sperm analysis), normal forms and plasma membrane integrity (evaluated under bright-field and fluorescence microscopy respectively), and DNA fragmentation (assessed by the Sperm-Halomax kit) were compared be-tween treatments. Sperm recovery rates were similar between SU and SLC but lower than SW. Sperm motility after thawing was lower in SU in comparison to SLC and SW, maybe due to the incomplete removal of seminal plasma before freezing. Sperm DNA fragmentation was lower in SU and SLC selection methods, particularly in SLC selected samples during the first 6 h of in-cubation. The remaining sperm parameters assessed were similar among treatments. In conclu-sion, SLC is more suitable than SW and SU to process stallion semen prior to freezing, in parti-cular when sperm DNA damage is suspected. Further studies are needed in order to determine the potential benefits of SU in samples where centrifugation is not necessary, such as epididymal sperm, ejaculate fractioning or post-thaw semen samples.Item Comparison of different sucrose-based extenders for stallion sperm vitrification in straws(Reproduction Domestic Animals, 2018) Consuegra González Cesar; Crespo Castejón, Francisco; Dorado Martín, Jesús; Ortiz Jaraba, Isabel; Diaz Jiménez, María; Pereira Aguilar, Blasa; Hidalgo Prieto, ManuelVitrification of sperm is based on high-speed freezing by direct exposure to liquid nitrogen using non-permeable cryoprotectants, mainly disaccharides; yet, the concentration of cryoprotectants has a species-specific effect on the sperm cell. The aim of this study was to assess different sucrose concentrations for stallion sperm vitrification. Semen samples (n = 9) were collected from three stallions, centrifuged and resuspended to a concentration of 50 × 106 sperm/ml in a base extender (INRA96 + 1% of bovine serum albumin) with three different sucrose concentrations (Molar): 20 mM (S1), 100 mM (S2), or 200 mM (S3). Then, sperm were filled in covered 0.25 ml straws and directly plunged into liquid nitrogen. For warming, 0.25 ml straw was pulled out the covering straw and immersed in 3 ml of INRA96 at 43°C, with gentle pipetting to accelerate the melting. Total (TM, %) and progressive sperm motility (PM, %) were analysed using computer-assisted sperm analysis. Plasma (PMI, %) and acrosome membrane integrity (AIS, %) were assessed under epifluorescence microscopy. Post-warmed sperm parameters were compared between treatments by ANOVA. S2 showed significantly higher values in comparison with S1 and S3 for TM (S2 = 54.7 ± 5.5a ; S1 = 29.1 ± 3.3b ; S3 = 28.6 ± 3.0b ; p < 0.001) and PM (S2 = 31.3 ± 3.8a ; S1 = 18.5 ± 2.6b ; S3 = 17.7 ± 2.9b ; p < 0.01), respectively. No significant differences were found among treatments for PMI (S2 = 70.3 ± 5.2; S1 = 67.4 ± 4.3; S3 = 70.0 ± 3.7) neither for AIS (S2 = 57.1 ± 3.9; S1 = 53.9 ± 4.2; S3 = 57.0 ± 7.9). In conclusion, a concentration of 100 mM sucrose is recommended for stallion sperm vitrification in straws.Item Vitrification of Large Volumes of Stallion Sperm in Comparison With Spheres and Conventional Freezing: Effect of Warming Procedures and Sperm Selection(Journal Equine Veterinary Science, 2019) Consuegra, Cesar; Crespo Castejón, Francisco; Dorado Martín, Jesús; Díaz Jiménez María; Pereira Aguilar Blasa; Arenas, R; Morrel, J M; Hidalgo Prieto ManuelStallion sperm was vitrified using straws in comparison with spheres and conventional freezing. Vitrification was performed plunging 30 mL of sperm (spheres) or 0.5 mL straws into liquid nitrogen (LN2) and conventional freezing using 0.5 mL straws frozen in LN2 vapors. Sperm vitrified in straws were submitted to different warming procedures (42C/20 seconds; 60C/15 seconds) and single-layer centrifugation (SLC). Total (TM, %) and progressive sperm motility (PM, %), plasma mem-brane (IMS, %) and acrosome integrity (AIS, %) were statistically compared between treatments (mean ± SEM). Significant higher values (P < .001) were obtained after vitrification using spheres in comparison with conventional freezing and vitrification in straws for TM (54.46 ± 3.2 vs. 36.47 ± 3.2 vs. 2.50 ± 1.2, %), PM (38.63 ± 3.4 vs. 15.11 ± 2.0 vs. 1.9 ± 0.9, %), IMS (65.40 ± 2.8 vs. 50.50 ± 2.8 vs. 21.63 ± 2.1, %), and AIS (48.89 ± 2.8 vs. 15.46 ± 1.7 vs. 4.69 ± 0.9, %). No differences were found between warming procedures. Single-layer centrifugation after warming at 42C/20 seconds ob-tained higher values (P < .05) than unselected samples for TM (32.52 ± 5.8%), PM (14.22 ± 2.8%), IMS (60.01 ± 3.2%), and AIS (44.5 ± 2.2%), whereas selection after 60C/15 seconds increased TM (23.11 ± 4.3%) and IMS (67.11 ± 3.9%). In conclusion, vitrification in spheres obtained better sperm quality than conventional freezing and vitrification in straws. Warming procedures did not affect the sperm quality but SLC could be a strategy to enhance the quality of the samples after sperm vitrification using 0.5 mL straws