Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

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16 t h International Congress on Animal Reproduction 78 Poster Abstracts unilateral growth of the head of epididymis. The animal presented right testicular enlargement, with higher temperature at the affected testis and lower testicular mobility. This animal had a history of haematuria and was treated with Pentabiotico® (penicillin) associated with sexual rest. After that the ram was reintroduced to breeding with 30 ewe and ten days later he showed the clinical signs of epididymitis. The pregnancy diagnose from these ewes by ultrasonography after 40 days showed 20 pregnant ewes (66,7%) of 30. In the andrological exam it was found many inflammatory cells and low motility. It was made specific bacteriological exam for Brucella sp, Mycoplasma sp., Ureaplasma sp and Campylobacter sp resulting negative. It was isolated only Actinobacillus semnis. The ovine production at the State of São Paulo, Brazil grows over the last 5 years. It has not many information about reproduction diseases in ram in this state. P151 Determination Sex and Scrapie Resistance Genotype 6f Preimplantation Caprine Embryos Guignot, F Physiologie de la Reproduction et des Comportement, INRA, France The aim of this study was to test the accuracy of sex and PrP genotype diagnosis after whole amplification of DNA extracted from biopsies obtained by microblade cutting of goat embryos, and to evaluate the viability of biopsied embryos after freezing / warming and transfer to recipients. Before genotyping, whole genome amplification was performed using Repli-g® kit. Sex diagnosis was done by PCR amplification of ZFX/ZFY and SRY sequences (duplex PCR) and PrP genotype determination was performed on five codons: 142, 154, 211, 222 and 240. Embryos were recovered at Day 7 after oestrus from superovulated goats. Blastocysts and expanded blastocysts were biopsied immediately after recovery and frozen by vitrification. Whole embryos were kept as control and also vitrified. After thawing, two embryos were transferred directly into each synchronized recipient. Transfer was performed on 28 recipients, 18 with biopsied embryos and 10 with whole embryos. At Day 21 and 42 after oestrus, pregnancy rate was assessed by progesterone RIA and ultrasonography respectively. PrP genotyping was performed on kids at birth. Pregnancy rates at Day 21, Day 42 and at kidding were not significantly different between biopsied (83, 39 and 39%, respectively) and control (100, 50 and 50%, respectively) embryos. The embryo survival rate was 25% (9 kids from 36 transferred embryos) and 35% (7/20) for biopsied and control embryos, respectively. At birth, all diagnosed sexes were right (9/9; 100%). Kid PrP genotyping and prediction were compared: among the 90 predetermined codons for PrP scrapie, 88 codons were accurately predicted (97.8%). Kid PrP profiles were in agreement with parental genotype. PrP genotyping results of biopsies from embryos who did not induce pregnancy was compared to parent genotypes. Among 270 analysed codons (27 biopsies), 8 discrepancies were found, so 97% of PrP genotype determination were concordant to parent genotype. Whole genome amplification with Repli-g® kit coupled with sex diagnosis and PrP genotype determination are very accurate techniques to genotype goat embryo before transfer. This technique will increase the efficiency and reduce the cost of selection process aimed at scrapie sensitivity alleles eradication in French goat population. Moreover, other target genes linked to other diseases or to production traits could be analysed on the same biopsy conferring more economical value to the multiple ovulation embryo transfer (MOET). P152 Cyclic female goats respond to males with an increase in LH secretion during the breeding season Hawken, P *; Esmaili, T; Jorre De St Jorre, T; Martin, G 1School of Animal Biology, University of Western Australia, Australia Introduction The male effect is currently only used during anoestrus because the long periods of elevated progesterone in cyclic females are presumed to block the response. However, in a recent study in sheep, we found that cyclic ewes respond to rams with an increase in LH secretion [1]. In the present study, we tested whether cyclic, female goats would respond to males with an increase in pulsatile LH during the early, mid- and late luteal phases of the oestrous cycle. Methods During May (breeding season; Southern Hemisphere) the oestrous cycles of 16 Australian Cashmere goats were synchronised using intravaginal progesterone pessaries. Pessary insertion was staggered to produce early luteal (EL; n=8) and late luteal phase groups (n=8). The late luteal phase group was further subdivided into mid-luteal (ML; n=4) and late luteal (LL; n=4) groups, based on differences in oestrous cycle length after progesterone withdrawal. Results Exposure to males stimulated an increase in LH pulse frequency in the EL (0.36 ± 0.06 versus 0.61 ± 0.08 pulses/h; P < 0.01) and LL groups (0.42 ± 0.08 versus 0.68 ± 0.09 pulses/h; P < 0.01). However, a significant increase was not observed in the ML group (0.33 ± 0.07 versus 0.45 ± 0.05 pulses/h; P > 0.1). Exposure to males caused an increase in mean concentrations of LH in the LL group (0.20 ± 0.05 versus 0.34 ± 0.05; P < 0.05) but not in EL (0.14 ± 0.03 versus 0.21 ± 0.05 ng/mL; P < 0.1) or ML groups (0.12 ± 0.02 versus 0.12 ± 0.03 ng/mL; P > 0.1). There was no effect of male exposure on LH pulse amplitude at any stage of the cycle (P > 0.1). Progesterone concentrations differed among all groups on the day of male exposure (EL: 3.50 ± 0.49; ML: 5.25 ± 0.47; LL: 0.72 ± 0.13 ng/mL; P ≤ 0.05) and declined significantly over the 12-h sampling period in the LL group (0.72 ± 0.13 versus 0.32 ± 0.05 ng/mL; P < 0.05). There was no change in this variable in the EL (3.5 ± 0.49 versus 2.96 ± 0.56 ng/mL; P < 0.1) or ML groups (5.25 ± 0.47 versus 5.99 ± 0.66 ng/mL; P > 0.1). Conclusion Exposure to males induced an increase in pulsatile LH secretion in female goats in the early and late luteal phases of the oestrous cycle. However, the high concentrations of progesterone during the mid-luteal phase appear to block any effect of the male on LH release. [1] Hawken PAR, Beard AP, Esmaili T, Kadokawa H, Evans ACO, Blache D, Martin GB. The introduction of rams induces an increase in pulsatile LH secretion in cyclic ewes during the breeding season. Theriogenology 2007;68:56-66. P153 Effect of the progestagen treatment length and different eCG dosages on the pregnancy rate of ewes after fixedtime superficial transcervical insemination Iwamura, J 1 *, Oba, E 1 ; Souza, MIL 2 ; Pampani, FE; Leal, LS 1 ; Pavão, G 1 ; Bittencourt, RF 1 1Department of Animal Reproduction and Veterinary Radiology, São Paulo State University, Brazil; 2 Center of Biological Science and Health , Mato Grosso do Sul Federal University, Brazil The aim of this study was to analyze the Santa Ines sheep’s fertility after estrous and ovulation synchronization with short, intermediate and long-term protocols with progestagen (Progespon®, Syntex, Argentina) and different eCG dosages (Foligon®, Intervet, Spain) through fixed-time superficial transcervical insemination. Fifty and three adult ewes were divided into 6 groups, according to the estrous synchronization protocols (P) employed. In the P1 and P2, the animals were synchronized with intravaginal sponges containing 60 mg of acetate of medroxyprogesterona (MAP) during 6 days plus 500 or 350 i.u. eCG at the sponge removal, respectively. P3 and P4 animals had the sponge removal after 9 days and received in this moment 500 or 350 i.u. eCG, respectively. In P5 and P6, the synchronization was accomplished by MAP administration over 13 days with an injection of 500 or 350 i.u. eCG, respectively, at sponge removal. The estrous detection was carried out twice daily by sexually experienced adult rams with proven high fertility, fitted with a marking harnesses, introduced in the flock at the sponge removal. The semen was collected by artificial vagina and the dilution (1/2) was carried out in skimmed milk. The superficial transcervical insemination was performed with 100µL of the diluted semen in fixed-time 48 hours after the sponges withdrawal. The pregnancy diagnosis was performed by ultrasonography exam, with a 5 MHz transducer, 60 days after the artificial insemination. The blood samples were collected from all ewes for progesterone determination by a direct solid-phase
16 t h International Congress on Animal Reproduction Poster Abstracts 79 radioimmunoassay (Coat-A-Count progesterone; Diagnostic Products Co., USA). The progesterone were measured in samples on the beginning treatment, before the sponge introduction, on the sponge removal day and 48h after the MAP removal. Data regarding the pregnancy rate and progesterone concentration were analyzed by using the qui-square test and the SNK test of the SAS statistical package (version 5.0, 1996). The fertility rate in the protocols P1, P2, P3, P4, P5 and P6 was of 30.0 %; 20.0%; 33.3%; 37.5%; 33.3% and 40.0%, respectively. No statistical difference for pregnancy rate was observed among protocols, hence we can conclude that length of progestagen treatment and the eCG dosages used had no influence on fertility rate. The authors suggest the necessity of more studies with a larger number of animals to verify the short-term estrus synchronization protocol effectiveness in Santa Ines ewes and the effect of the superficial cervical artificial insemination on the pregnancy rates. P154 Seasonal changes in physiologic and biochemical parameters of Iranian Abadeh breed buck semen Jelodar, G *, Razmi, N Department of Physiology, Shiraz University, Islamic Republic of Iran Introduction Goat is a seasonal breeder animal,the aim of this study was to evaluate seasonal changes in physiologic and biochemical semen characteristics of native Abadeah breed buck goats. Methods Five Abadeah goats (3-4 years old) were trained to serve the artificial vagina. Semen collection was performed every 2 weeks, commencing in October (at onset of autumn) 2005 to September (end of summer) 2006. Semen ejaculates were evaluated for volume, sperm concentration, individual motility and the percentage live sperm. Moreover, changes in the seminal plasma proteins of the goats were also recorded and considered at the end of semen collection whit providing electrophorogram (SDS PAGE Electrophoresis) of seminal plasma proteins. Result Semen of superior quality and quantity was especially collected in late summer and throughout autumn (seasonal breeder). during the autumn bucks recorded the highest (P
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