Plasma IGF1 and 17β-Estradiol Concentrations During the Follicular Wave in Llamas

The aim of this study was to characterize the temporal association between follicular waves and circulating concentrations of 17β-estradiol (E2) and IGF1 in llamas. Follicular waves could be clearly divided in three phases: growth, plateau and regression; with a mean duration of 18.8 ± 0.32 days. All follicular waves showed overlapping, so that as one dominant follicle was regressing, another one was growing. E2 plasma concentration showed a wavelike pattern, similar to that followed by the dominant follicle; reaching its maximum concentration at the end of the growth phase and decreasing at the end of the plateau phase. IGF1 also showed variations during the follicular wave. It tended to increase during the growth phase and decreased toward Days 14 and 16. IGF1 reached its maximum concentration before E2 did (5 ± 0.8 vs. 7.2 ± 0.5 days after wave emergence) and before the maximum follicular diameter was attained (10.2 ± 0.46 days after wave emergence). Both hormones started to rise again in coincidence with the development of a new follicular wave. The observed profiles allow to suggest that IGF1 could have a role on folliculogenesis and ovarian steroideogenesis in llamas, as reported for other species.Fil: Gallelli, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Bianchi, Carolina Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Zampini, Enzo German. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Aba, Marcelo Alfredo. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentin

Air-drying llama sperm affects DNA integrity

The objective of this study was to evaluate the effects of air-drying preservation on llama sperm DNA. Semen collections were carried out using electroejaculation under general anesthesia. A total of 16 ejaculates were processed from 4 males (n = 4, r = 4). Each sample was diluted 4:1 in a collagenase solution in TALP media, then incubated and centrifuged at 800 g for 8 min. The pellet was re-suspended to a concentration of 20 million sperm/ml in TALP. Then the samples were placed onto sterile slides forming lines and were left to dry under laminar flow for 15 min. After this, the slides were placed into Falcon centrifuge tubes and kept at 5°C. Sperm characteristics (motility, membrane function, viability and morphology) were evaluated in raw semen and in the air-dried samples kept at 5°C for 30 min. DNA evaluation (integrity and degree of chromatin condensation) was carried out in raw semen and in the air-dried samples after 30 min, 7, 14, 21, 30, and 60 days after preservation. To compare raw semen to the air-dried samples, a Wilcoxon test was used for all sperm characteristics except for DNA, where a paired Student t-test was applied. A split plot design was used to compare chromatin condensation between the different periods of preservation and a Kruskal Wallis test was used to compare DNA integrity. Motility, membrane function, viability and sperm with intact DNA decreased in the air-dried samples (p 0.05). No significant differences were observed in the percentage of sperm with condensed chromatin between the different periods of preservation (p > 0.05). On the other hand, a significant decrease in the percentage of sperm with intact DNA was observed as from day 7 of preservation (p < 0.05). In conclusion the air-drying process has a negative effect on llama sperm DNA, hence the media used will need to be improved to protect DNA and be able to implement this technique in this species.Fil: Carretero, Maria Ignacia. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Chaves, María Graciela. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Arraztoa, Claudia Cecilia. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Fumuso, Fernanda Gabriela. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gambarotta, Mariana Carla. Universidad de Buenos Aires; ArgentinaFil: Neild, Deborah Margarita. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentin

Uterine and corpus luteum blood flow evaluation prior to uterine flushing in llama embryo donors

The aim of this study was to assess the uterine blood flow (UBF) and corpus luteum blood flow (CLBF) in llamas 8 days post-mating, using color-Doppler ultrasonography (CDU), to determine the possible relationship between vascularization and the presence of an embryo. Adult females (n = 25) were used to monitor ovarian dynamics by palpation and transrectal ultrasonography until detection of a ≥6 mm growing follicle. Females were randomly assigned to one of two groups: Group I (n = 19), were mated and ovulation was induced by a single dose of buserelin (GnRH analog) that same day (Day 0); and Group II (n = 6), only ovulation was induced (control). On Day 8, UBF and CLBF were evaluated transrectally in both groups. The color-flow images obtained were analyzed with Image J1.52a software to determine the vascularization area and the percentage of corpus luteum with blood flow emission (CLBF%) together with the percentage for each uterine horn (UBF%). Statistical analysis was performed using an ANOVA test. In Group I, uterine flushing was performed to obtain the embryos, thus dividing the females into Group I+ (n = 10), when an embryo was recovered and Group I- (n = 9), when no embryo was recovered. Embryo recovery rate was 52.63% (10/19). In Group I+, UBF% was significantly higher compared to Group I- and Group II (P 0.05). In conclusion, it is possible to detect a local increase of UBF in the presence of an embryo on day 8 post-mating in llamas. This could be useful to achieve an early pregnancy diagnosis or to decide whether to carry out the uterine flushing in a llama embryo transfer program.Fil: Zampini, Enzo German. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; ArgentinaFil: Gallelli, Maria Florencia. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; ArgentinaFil: Chaves, María Graciela. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Neild, Deborah M.. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Miragaya, Marcelo H.. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Trasorras, Virginia Luz. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentin

Transfer of cooled llama embryos obtained from synchronized females

This study evaluated the efficiency of a synchronization protocol based on GnRH and PGF2α on embryo donor llamas for fixed timed mating and assessed the viability of embryos maintained at 5 ◦C and 15 ◦C for 24 h, using the Equitainer® and the Botu-BOX® as cooling devices respectively. Llamas were divided into four follicular wave groups: growth, dominance, static and regression; they received a GnRH analogue on day 0 followed by a second dose plus cloprostenol on day 8 and 15 and mating was indicated in females with a follicle ≥ 6 mm. Embryos were recovered 8 days post mating. Synchronization rate was 80% for the treated embryo donors, with no significant differences among groups. Uterine flushing was performed in 70% of the treated females (87.5% of mated llamas) and an embryo was recovered in 50%. Fourteen embryos were assigned randomly to 5 ◦C (Equitainer® group) and 15 ◦C (Botu-BOX® group) preservation for 24 h to be transferred later. In the Equitainer® group, we obtained 14% pregnancies and a female offspring was born. In the Botu-BOX® group, 28% resulted pregnant but subsequently pregnancies were lost. This protocol was effective for synchronizing follicles in growth phase in 80% of embryo donor llamas. In addition, cooling llama embryos using the Equitainer® and the Botu-BOX® as cooling devices to 5 ◦C and 15 ◦C respectively, preserves its morphology and viability for 24 h.Fil: Zampini, Enzo German. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Fernanda Veiga, María. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Arraztoa, Claudia Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; ArgentinaFil: Gallelli, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Moncalvo, Carla Evangelina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Neild, Deborah Margarita. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Trasorras, Virginia Luz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentin

In vitro inhibitory activity of IgY antibodies against Salmonella ser. newport isolated from horses

Equine salmonellosis is caused by several Salmonella serotypes, including Salmonella Newport, which cause enterocolitis and diarrhea. Treatment usually includes the administration of antibiotics. However, since multidrug-resistant Salmonella is commonly detected, alternative options to control the pathogen are needed. One of these options is the use of specific egg yolk antibodies (IgY) for passive immunotherapy. Thus, the aim of our work was to produce IgY antibodies against an equine S. Newport strain and assess their in vitro inhibitory activity. To this end, laying hens were immunized with an inactivated S. Newport strain by using either Freund's or Montanide adjuvant and egg yolk extracts were obtained. The levels of specific IgY antibodies against Salmonella in sera and egg extracts were determined by dot-blot and microagglutination. Besides, the IgY extracts were characterized by total protein analysis, SDS-PAGE, Western Blot, and inhibition of bacterial motility. IgY extracts showed high purity (87.7 to 91.8 %), high microagglutination titers, and the ability to inhibit the motility of the bacterium. The results using Montanide were similar to those using the traditional Freund's adjuvant. Thus, Montanide may also be a good adjuvant to produce IgY. IgY-technology represents a potential tool for the control of salmonellosis in horses.Instituto de PatobiologíaFil: Bustos, Carla Paola. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Bustos, Carla Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bustos, Carla Paola. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Enfermedades Infecciosas; ArgentinaFil: Leiva, Carlos Leónidas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Leiva, Carlos Leónidas. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Bioestadística; ArgentinaFil: Guida, Nora. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Enfermedades Infecciosas; ArgentinaFil: Chacana, Pablo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Chacana, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Corpus luteum vascularization during the maternal recognition of pregnancy in llamas (Lama glama)

The aim of this study was to characterize corpus luteum vascularization and its association with plasma progesterone concentration in early stages of pregnancy, when maternal recognition of pregnancy is expected to occur. In all animals, both plasma progesterone concentration and corpus luteum vascularization increased from Day 6 to Day 8 post-mating and afterwards in non-pregnant llamas they started to decrease to reach basal levels around Days 12 to 14 post-mating, while in pregnant animals, both variables remained elevated until the end of the study. A lineal positive relationship between corpus luteum vascularization and plasma progesterone concentration was observed in pregnant (r2 =.46, p <.0001) and non-pregnant llamas (r2 =.66, p <.0001). Pregnant animals showed higher plasma progesterone concentration and corpus luteum vascularization than the non-pregnant ones from Day 12 post-mating until the end of the study (p ˂.05 and p ˂.01, respectively). These results suggest that maternal recognition of pregnancy should occur before Day 12 post-mating in order to expand luteal lifespan, maintaining corpus luteum vascularization and progesterone production. Also, the assessment of CL vascularization area could be a useful and non-invasive method for early pregnancy diagnosis due to its association with plasma progesterone concentration.Fil: Gallelli, Maria Florencia. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bianchi, Carolina Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Fisiopatología. Laboratorio de Endocrinología; ArgentinaFil: Zampini, Enzo German. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Trasorras, Virginia Luz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentin

Porcine sperm vitrification I: cryoloops method

The aims of this study were to evaluate porcine sperm vitrification in cryoloops, with and without two different cryoprotectants and assess two warming procedures. Extended (n = 3; r = 4) and raw (n = 5; r = 2) semen was diluted in media without and with cryoprotectants (4% dimethylformamide and 4% glycerol) to a final concentration of 20 × 106 spermatozoa ml−1 and vitrified using the cryoloops method. Two warming procedures were evaluated: rapid method (30 s at 37°C) and an ultra-rapid method (7 s at 75°C, followed by 30 s at 37°C). Total motility (phase contrast), sperm viability (6-carboxifluorescein diacetate and propidium iodide stain), membrane function (hypo-osmotic swelling test), acrosome integrity (phase contrast), chromatin condensation (toluidine blue stain) and chromatin susceptibility to acid denaturation (acridine orange stain) were evaluated before and after vitrification and analysed using Friedman's test. In all media, the only seminal parameters that were maintained after vitrification were chromatin condensation and integrity. Vitrification of porcine spermatozoon using cryoloops, both in the presence or absence of cryoprotectants and independent of the warming procedure used, permits conservation of sperm chromatin condensation and integrity. It would be interesting to further verify this by producing porcine embryos using vitrified spermatozoon with intracytoplasmic sperm injection.Fil: Arraztoa, Claudia Cecilia. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Chaves, María Graciela. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Trasorras, Virginia Luz. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Péndola, Carlos Horacio. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; ArgentinaFil: Neild, Debora Margarita. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Area de Teriogenología; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentin

Development of an artificial insemination protocol in llamas using cooled semen

The objective of this study was to design an AI protocol using cooled semen to obtain pregnancies in the llama. Each raw ejaculate was subdivided into four aliquots which were extended 1:1 with: (1) 11% lactose-egg yolk (L-EY), (2) Tris-citrate-fructose-egg yolk (T-F-EY), (3) PBS-llama serum (S-PBS) and (4) skim milk-glucose (K). Each sample reached 5°C in 2.5h and remained at that temperature during 24h. Percentages of the semen variables (motility, live spermatozoa) in ejaculates and samples cooled with L-EY were significantly greater than those obtained when cooling with the other extenders; therefore this extender was used (1:1) for all inseminations. Females were randomly divided into four groups (A-D) according to insemination protocol. Group A: females were inseminated with a fixed dose of 12×10 6 live spermatozoa kept at 37°C. Group B: females were inseminated with a fixed dose of 12×10 6 live spermatozoa, cooled to 5°C and kept for 24h. Group C: females were inseminated with the whole ejaculate (variable doses), cooled to 5°C and kept for 24h. These groups (A-C) were inseminated between 22 and 24h after induction of ovulation. Group D: females were inseminated with the whole ejaculate (variable doses), cooled to 5°C, kept for 24h and AI was carried out within 2h after ovulation. Pregnancy rates were 75%, 0%, 0% and 23% for groups A, B, C and D respectively. These results indicate that it is possible to obtain llama pregnancies with AI using cooled semen and that the success of the technique would depend on the proximity to ovulation.Fil: Giuliano, Susana María. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Chaves, María Graciela. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Trasorras, Virginia Luz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Neild, Debora Margarita. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Director, Ariel. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Pinto, Marcelo R.. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentin

In vitro equine embryo production using air-dried spermatozoa, with different activation protocols and culture systems

The aim of this work was to evaluate the use of air-dried spermatozoa for in vitro production of equine embryos and verify if sperm extract activation and in vivo culture improve in vitro embryo production. Cooled spermatozoa (control) and air-dried spermatozoa stored for 2, 14 or 28 days were used for ICSI sperm extract, or ionomycin was used for oocyte activation, and embryos were in vitro or in vivo (in mare′s oviduct) cultured for 7 days. With in vitro culture, cleavage rate was higher when activating with sperm extract (P 0.05). Blastocysts were obtained with cooled spermatozoa, and morulae were achieved using in vivo culture with 28-day storage spermatozoa and ionomycin-activated oocytes. When in vivo culture was performed, sperm DNA fragmentation was assessed using the sperm chromatin dispersion test and did not show statistical correlation with cleavage nor embryo recovery rates. In conclusion, equine embryos can be produced using air-dried spermatozoa stored for several weeks. Sperm extract activation increased cleavage rates but did not improve embryo development. In vivo culture allowed intrauterine stage embryos to be achieved.Fil: Alonso, Ana Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Baca Castex, Clara. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Ferrante, Alejandro Antonio. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Pinto, Marcelo Raul. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Castaneira, Catalina. Universidad Nacional de Rio Cuarto. Facultad de Agronomia y Veterinaria. Departamento de Producción Animal. Laboratorio de Reproducción Equina; ArgentinaFil: Trasorras, Virginia Luz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Gambarotta, Mariana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; ArgentinaFil: Losinno, Luis. Universidad Nacional de Rio Cuarto. Facultad de Agronomia y Veterinaria. Departamento de Producción Animal. Laboratorio de Reproducción Equina; ArgentinaFil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias; Argentin