Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

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16 t h International Congress on Animal Reproduction 166 Poster Abstracts P420 Seasonal variations in the sperm parameters in dairy bulls regularly used for artificial insemination in Algeria Iguer-Ouada, M 1,2 *, Ayad, H 2 , Abbas, G 2 , Azzeradj, M 2 1Department of Organisms and Populations, Faculty of Nature and Life Sciences, University of Bejaia Algeria, Algeria; 2 Department of Organisms and Populations, University Abderahmane Mira, 06000, Bejaia, Algeria, Algeria The present work is the first report describing bull semen quality variations during the different annual seasons in this part of the word (North Africa). Data from 503 ejaculates collected during 14 months from 06 mature Holstein bulls were analyzed and different semen parameters were evaluated. Semen volume was measured using calibrated plastic vial, and gametes concentration was determined using a haemocytometer. Mass motility of semen was graded from a 0–5 scale, based on the appearance of waves and swirls created by sperm movement when visualized by keeping one drop of semen on a glass slide, without cover slip, under low power microscopic magnification (10×). The individual motility of freshly diluted semen was assessed after covering a semen drop on a glass slide with a thin cover slip under high power magnification (40×). The individual motility was recorded as the percentage of progressive motile sperm. Semen concentration and motility percentage showed a parallel evolution with a regular decrease form the spring season were the highest values were observed to reach the lowest values in the summer season. For motility percentage the values were 55.96 ± 1.94, 54.07 ± 1.9, 50.82 ± 2.46 and 37.33 ± 7.12 respectively for spring, winter, autumn, and summer seasons (Mean ± SE). The semen volume showed an inverse evolution increasing from winter (6.29 + 0.19 ml) to reach maximum values in summer (7.1 + 0.69 ml). It was concluded in the present work that season affected significantly different bull semen parameters. However, in one hand the confounding effects of ambient temperature and photoperiod on the semen quality variation in North Africa region needs further examinations as well as the impact of these variations in term of fertility. P421 Hyperthermia is more important than hypoxia as a cause of disrupted spermatogenesis Kastelic, J 1 *; Wilde, R 1 ; Bielli, A 2 ; Genovese, P 2 ; Bilodeau-Goeseels, S 1 ; Thundathil, J 3 1Agriculture and Agri-Food Canada, Lethbridge Research Centre, Canada; 2Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay; 3Faculty of Veterinary Medicine, University of Calgary, Canada Mammalian testes operate on the brink of hypoxia; furthermore, it is believed that hyperthermia-induced disruptions in sperm quality and production are primarily due to hypoxia. The objective of this study was to determine the relative effects of hypoxia versus hyperthermia on sperm quality and production. We tested the hypothesis that hypoxia would disrupt sperm quality and production, whereas hyperoxia would prevent hyperthermia-induced reductions in sperm quality and production. Forty-eight CD-1 mice (approximately 50 days of age), were randomly allocated into six groups and exposed to environmental conditions of 20 vs 36 °C and oxygen (O 2 ) concentrations of 13, 21, or 95% (2 x 3 factorial experiment) for 12 hours on two occasions (separated by 12 hours at 20 °C and 21% O 2 ), and euthanized (CO 2 ) 14 or 20 days later. One cauda epididymis was minced and placed in PBS for 30 minutes; sperm motility was assessed subjectively, and smears were prepared and stained with Eosin Y. One testis was fixed in Bouins (24 hours), transferred to 70% alcohol, and sections prepared and stained with hematoxylin and eosin. Morphological assessments were done ‘in the blind’ on semen smears (200 sperm/mouse) and testicular sections (30 videocamera fields/mouse). Data are mean±SD (combined for both days). There were primarily main effects of temperature; mice exposed to 20 vs 36 °C had differences in testis weight (110.4±14.3 vs 101.3±17.6 mg, P
16 t h International Congress on Animal Reproduction Poster Abstracts 167 under extensive rearing with natural mating and are characterized by lack of selection, poor record keeping and infrequent veterinary assistance. Besides, breeding bulls are selected based mainly on phenotype, neglecting the importance of the breeding soundness evaluation (BSE) (1). Materials and Methods A single BSE was performed in 38 sires placed in 26 beef cattle farms managed under extensive rearing and continuous mating from the south area of Costa Rica. Bulls belonged to Brahman (n=8), Simmental x Brahman crosses (n=17), Simmental (n=6), Brown Swiss x Brahman (n=3) and other crosses (n=4). These bulls were under single sired mating breeding with 24.0±13.6 (5-73) adult cows. After the BSE, bulls were classified as Sound: Bulls that fitted the physical exam, with a scrotal circumference (SC) according to breed and age standards (2), healthy reproductive organs and without deviations in the spermiogramme. Deferred: Sires who did not meet the above requirements but with a favorable prognosis for recovering. Unsound: Serious clinical and spermiogramme abnormalities compromising gravely their potential breeding efficiency. Simultaneously, the conception rate (CR) was evaluated in the herds by rectal palpation and ultrasound. Thereafter, the cows were ranked as pregnant or not, being the cycling activity determined by the presence of a corpus luteum. Results and Discussion Mean SC (cm) and age (yrs) for sound (n=23), deferred (n=5) and unsound (n=10) sires were 37.4±3.3 and 4.5±2.4, 39.0±4.4 and 4.7±2, 37.6±5.2 and 4.2±2 respectively. In the unsound group, one bull had low SC and the remnant had clinical and spermiogramme deviations typical of testicular degeneration. The CR calculated solely upon the cycling cows was 82.5%±12.1 (64-100) for sound, 77.4%±27.2 (36-100) for deferred and 28.4%±30.5 (0-79) for the unsound sires. The CR was p0.05 when comparing the sound vs. the deferred group. Given the conditions of cattle rearing prevailing in the tropics, the BSE is an efficient tool in order to identify those bulls with an unsound andrological status. The presence of those sires in the herd, contributes to impair seriously the productive proficiency of the beef cattle system. P424 Effect of epididymis storage temperature and cryopreservation on mitochondrial potential, membrane integrity and motility of bovine epididymal sperm Nichi, M 1,2 *, Rijsselaere, T 3 , Goovaerts, IGF 2 , Van Soom, A 3 , Barnabe, VH 1 , De Clercq, JPB 2 , Bols, PEJ 2 1Department of Animal Reproduction (VRA), Faculty of Veterinary Medicine and Zootechny (FMVZ), University of São Paulo (USP), Brazil; 2 Laboratory of Veterinary Physiology, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, Belgium; 3 Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Belgium Introduction Cryopreservation of sperm recovered from epididymides is potentially a useful tool in case of unexpected death of animals with high genetic value, species endangered of extinction, or when the collection of sperm by other means is not possible. Studies indicate that maintaining the epididymis at a lower temperature during storage and transport improves the quality of the retrieved sperm. The objective of this experiment was to study whether the storage temperature of the epididymis following slaughter influences sperm membrane resistance against lipid peroxidation, resistance to cryopreservation, and fertilizing capacity, and if the percentage of cytoplasmic droplets would play a role in this event. Methods Thirty two epididymides (16 bulls) were collected after slaughter and divided into two groups, stored at 4 and 34ºC for 2-3 hours, after which semen was collected from the caudae epididymides. The epididymal sperm was then evaluated using CASA, and an aliquot was stained with SYBR14/PI/JC1 to evaluate membrane integrity and mitochondrial membrane potential. Sperm was then frozen using an automatic device. Post thaw sperm was also analyzed using CASA and SYBR14/PI/JC1. Results A marked negative effect of cryopreservation on sperm motility was observed in the samples collected from epididymides stored at 4ºC. Significant effects of temperature of epididymal storage and of cryopreservation were observed on motility parameters as well as on cell viability and mitochondrial activity. Pre-cryopreservation sperm motility, progressive motility, and velocity were higher in samples from epididymides stored at 4 o C, while post-thaw sperm motility, progressive motility, and velocity did not differ between samples from epididymides stored at 4 or at 34 o C. Although the drop on motility was not evident on sperm samples collected from epididymides stored at 34ºC, the effect of temperature before cryopreservation may have influenced those results. Strong correlations (r>0.6) were found for samples collected from epididymides stored at 34ºC between pre-freeze sperm motility indexes and mitochondrial potential and between post-thaw sperm motility and membrane integrity. Conclusions Results indicate that the conditions in which epididymides are handled after cryopreservation may influence postthaw sperm quality, especially due to an impaired mitochondrial potential in sperm samples collected from epididymides stored at higher temperatures. This effect may have caused a higher susceptibility to membrane damages due to cryopreservation. P425 Assessment of spermatozoal characteristics in extended boar semen incubated at 18 o C using flow cytometry Niżański, W 1 *; Partyka, A 2 ; Dubiel, A 1 ; Łukaszewicz, E 2 1Department of Reproduction, University of Environmental and Life Sciences in Wrocław, Poland; 2 Department of Poultry Breeding, University of Environmental and Life Sciences in Wrocław, Poland Extended liquid boar semen is commonly used for insemination for up to 5 days after collection. A litter size and farrowing rates are reduced when using semen stored > 3 days. The aim of our study was to estimate changes of spermatozoal characteristics in extended boar semen incubated at 18oC for 240 hrs using fluorescent staining and flow cytometry. The study was carried out on 35 ejaculates collected from 7 boars. Semen samples were extended in Safe Cell+ diluent (IMV Technologies). Spermatozoal characteristics were evaluated at 24, 48, 96, 168 and 240 hrs after collection. Spermatozoal viability was determined by SYBR-14/PI fluorescent staining. The DNA status of spermatozoa was assessed using the metachromatic properties of acridine orange (AO) in the sperm chromatin structure assay (SCSA). DNA fragmentation index (DFI) and high DNA stainability (HDS) were evaluated. Acrosomal integrity was measured by Pisum sativum (PSA) agglutinin labeled by a fluorescent probe that stains acrosomereacted- or damaged spermatozoa only. The mitochondrial function of spermatozoa was evaluated with Rhodamine 123 (R123). Samples were analysed in a FACSCalibur flow cytometer (Becton Dickinson, San Jose, CA). The percentage of live spermatozoa did not change significantly during the 168 hrs of storage. A significant increase of the dead spermatozoa percentage (p
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Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

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