Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

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16 t h International Congress on Animal Reproduction 108 Poster Abstracts problem mares, one may conclude that uterine cytology is the first choice exam to detect endometritis in mares. Acknowledgements: FAPESP 05/53112-4 and FAPESP 06/59003-5. P241 Sperm morphology variation of stallion ejaculates in different breeding seasons Morrell, JM*, Dalin, AM and Rodriguez-Martinez, H Division of Reproduction, Dept of Clinical Sciences, SLU, Box 7054, 75007 Uppsala, Sweden Previous reports have described seasonal variation in the proportion of stallion spermatozoa with normal morphology but there is a paucity of data on variation between different years. The present report is a retrospective analysis of morphology evaluations from 8 stallions over several years. The stallions were all from a commercial stud (Flyinge AB, Flyinge, Sweden). Three or four ejaculates were collected from each stallion under a three-week period in June 2006. Aliquots of each ejaculate were used to prepare air-dried stained smears or fixed in formol saline for later evaluation as wet smears, using the procedure described by Lagerlöf. The evaluation of sperm morphology was performed by skilled, experienced technical personnel at SLU. The proportion (%) of morphological spermatozoa was estimated and used to establish a 95% confidence interval (CI) for normal morphology for these stallions. Analyses were also available for individual ejaculates from the same stallions in previous years and in 2007. The results from other years were compared with the 95% CI for 2006: if the results lay within the 95% CI, they were considered not to be significantly different from the 2006 values; if they lay outside the 95% CI, they were considered to be significantly different. Therefore, of the 21 evaluations made in years other than 2006, 12 were outside the 95% CI and were considered to be significantly different from the 2006 means. Of these, 10 were higher than the 95% CI, whereas two were lower. However, nine ejaculates were not statistically different from the 2006 mean values. Interestingly, stallion R showed a steady decrease in the proportion of spermatozoa with normal morphology over 13 years. In conclusion, considerable variation in normal morphology occurs between ejaculates from any individual stallion in different breeding seasons, and this difference varies among stallions. Acknowledgments: Funded by the Swedish Equine Research Organisation, Stockholm. We thank Karen Selin-Wretling and Annika Rikberg for their skilled sperm morphology analysis, and Flyinge AB for semen samples. P242 Impact of a single administration of hcg on free thyroid hormone levels in mares at oestrus Mutinati, M*, Rizzo, A; Nicassio, M; Matarrese, R; Spedicato, M; Minoia, G; Lacalandra, GM; Sciorsci, RL Department of Animal Production, Faculty of Veterinary Medicine, Bari, Italy Human chorionic gonadotropin (hCG) is a glycoprotein hormone sharing a high structural resemblance with the molecule of the thyroid stimulating hormone (TSH). Furthermore, the extracellular domain of the LH/CG receptor and TSH receptor, share 45% homology. The aim of the study was to evaluate the effect of a single intra-muscular administration of hCG, on serum concentrations of free triiodothyronine (fT3) and free thyroxine (fT4) in mares. Twenty healthy, adult mares in oestrus, were divided in two groups: group A consisting of 10 subjects, treated with 5 ml of sterile saline solution, (NaCl 0.9%); group B consisting of 10 subjects, treated with 4000 I.U. of hCG (Chorulon®, INTERVET). All mares underwent 7 blood collections: T1, contextually to drug administration; T2, T3, T4, 2, 6 and 24 hours after drug administration, respectively; T5, T6, 3 and 6 days after drug administration. All mares were inseminated with refrigerated semen belonging to the same, proved stallion, soon after T3 and underwent a pregnancy diagnosis 14 days after insemination. Dosages of fT3 and fT4 were performed on all the blood samples collected, with immunoenzymatic competitive kits (EIA WELL®, RADIM, Pomezia, Italy). Data were statistically analyzed with chi squared, one way ANOVA and Student’s t-test. The conception rate of group A was 40%, whereas in group B it reached 80% (p
16 t h International Congress on Animal Reproduction Poster Abstracts 109 Miniature mares are a potential model for comparative studies in folliculogenesis within and among species. P244 The effect of interval from mating to ovulation on pregnancy rates and incidence of intra-uterine fluid in the mare Newcombe, JR 1 *; Cuervo-Arango, J 2 1Equine Fertility Unit, Warren house farm, Brownhills, UK; 2 Royal Veterinary College, Department of Veterinary Clinical Science, University of London, UK Introduction Persistent mating-induced endometritis (PMIE) is known to be one of the main factors affecting pregnancy rates in the mare. There has been extensive research on causes and treatments of PMIE, however the interval from mating to ovulation (IMO) has never been considered as a possible factor affecting the outcome of PMIE and pregnancy rates. It is thought that natural cover is best performed close to ovulation (0-48 h prior to ovulation) in order to achieve optimum pregnancy rates. However, to date it is not known whether mating “problem mares” at that interval will be detrimental for pregnancy rates (PR). The objectives of this study were: a) to assess the effect of IMO on PR and on the incidence of intra-uterine fluid (IUF) 72 h post-mating (PM); and b) to determine the effect of free IUF 72 h PM on PR. Material and methods A total of 439 cycles from 315 Thoroughbred mares were analysed. Mares were examined by transrectal ultrasonography every other day until the mare was free from IUF (up to 96 h from mating). Pregnancy diagnosis was preformed by ultrasound 12-13 days post-ovulation and rechecked by 30 days. For each mare, age, date of mating, name of stallion used, number of services, IMO and depth of IUF 72 h PM were recorded. For data analysis, three groups of IMO were used (0-48, 48-96 and 96-144 h). A statistical general model of analysis of variance was used to assess the effect of stallion, IMO, age and presence of IUF 72 h PM on PR. Chi-square test was used to analyse the association among IMO, PR and IUF 72 h PM. Results Stallion, age and IMO had no effect (P > 0.05) on overall (all services) PR. Pregnancy rates in mares with IUF 72 h PM was reduced by 17.7 % (P < 0.01). Mares mated at the interval 0-48 h preovulation were more likely (P < 0.01) to have IUF 72 h PM than at 48-96 and 96-144 h (68, 42 and 41 % respectively). Pregnancy rates on first service were (69.3, 73.3 and 68.4 % for intervals 0-48, 48-96 and 96-144 h respectively, P > 0.05). However, on second service (n = 88 mares) they were lower (P < 0.05) in mares mated 0-48 h (54.5 %) than in those mated 48-96h before ovulation (79.1 %) . Conclusion Incidence of IUF 72 h PM is highest in mares mated 0-48 h before ovulation. Mating 0-48 h before ovulation is detrimental for PR in mares which failed to conceive in first service. It appears that in “problem mares” with delayed uterine clearance, a longer IMO gives the mare an “extra” time to clear the uterine inflammation before the cervix closes under progesterone influence. P245 Apoptotic markers can be used to forecast the freezeability of stallion spermatozoa Ortega Ferrusola, C 1 *, Gallardo Bolaños, JM 1 , Gonzalez Fernandez, L 2 , Rodriguez-Martinez, H 3 , Tapia, JA 2 , Pena, FJ 1 1Reproduction, University of Extremadura, Spain; 2 Physiology, University of Extremadura, Spain; 3 Reproduction, SLU, Sweden In an attempt to identify valuable markers for potential freezeability of the equine spermatozoa, three ejaculates were collected from five Andalusian stallions and frozen using a standard protocol. Before freezing, three apoptotic cell markers were studied by flow cytometry (early changes in sperm membranes, mitochondrial membrane potential and caspase activity). Post-thaw, spermatozoa were again evaluated for these parameters and for sperm kinematics using CASA. Receiving operating system curves were used to evaluate the relative value of the apoptotic markers herein studied, as forecast for potential freezeability. From all parameters studied, the outcome of JC-1 (as proportion of spermatozoa showing simultaneously orange and green fluorescence) had the highest diagnostic power. For potentially bad freezers (less than 25% of intact spermatozoa post-thaw), the significant area under the ROC-curve was 0.985, with a 100% sensitivity and 99.8% specificity for a cut off value of 55.7. P246 Effect of altrenogest-treatment in late pregnant-mares on parturition, neonatal adaptation and adrenocortical function in new-born foals Palm, F 2 *; Neuhauser, S 1 ; Ambuehl, F 1 ; Moestl, E 3 ; Aurich, C 1 1Centre for Artificial Insemination and Embryo Transfer; 2 Clinic for Obstetrics, Gynaecology and Andrology; 3 Institute for Biochemistry, University of Vet. Sciences, Vienna, Austria In mares with compromised pregnancies, treatment with the synthetic gestagen altrenogest (allyltrenbolon) is common to maintain gestation until term. However, there are no controlled studies on the effects of altrenogest administration during late pregnancy on parturition and development of the foals in the immediate postpartum period. In our study, pony mares with normal pregnancies were treated with altrenogest (0.088 mg/kg bwt; n=6) from day 280 of gestation until spontaneous parturition or were left untreated as controls (n=7). Stages of labour and clinical parameters were analysed and blood samples were collected from immediately after birth until 48 hours post natum for analysis of cortisol concentration, haematolgical and acid base parameters. In order to assess adrenocortical function, an ACTH stimulation test with 0.125 mg ACTH (Synacthen, Novartis) was performed on days 1 and 5 after birth and cortisol release calculated as area under the curve for the time period 0 to 120 min after cortisol injection. Altrenogest-treated mares tended to have a shorter gestation length (320±4 days) than control mares (327±2 days) and showed a prolonged second stage of labour (p
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