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Ítem Acceso Abierto Alpaca embryo transfer on a private Canadian farm(Canadian Veterinary Medical Association, 2018) Palomino Cano, Jesús Manuel; Jones, Lori; Vanhanen, Tom; Mastromonaco, Gabriela F.; Busato, Rachel; Adams, Gregg P.This study evaluated the feasibility of using an embryo transfer protocol in an alpaca farm in Canada. Alpaca donors and recipients were synchronized with 2 doses of gonadotrophin-releasing hormone (GnRH), 12 days apart. In donors (n = 5), superstimulation was induced with follicle stimulating hormone (FSH) given daily (40 mg) for 5 days beginning 2 days after the second GnRH treatment. Cloprostenol was given on the last day of FSH, the donors were bred 2 days later, embryos were collected 7 days after breeding. In recipients (n = 8), the second dose of GnRH was given the day before donor mating, and embryos were transferred on the day of donor collection. On average (± SEM), 5.2 ± 1.4 corpora lutea were detected and 2.5 ± 1.2 transferable embryos were collected in the donors. A mature corpus luteum was detected in 6/8 synchronized recipients and a single embryo was transferred to each. One recipient alpaca became pregnant and delivered a healthy baby 349 days after embryo transfer. This is the first report of successful embryo transfer in alpacas in Canada.Ítem Solo Metadatos Cryopreservation of bison semen without exogenous protein in extender and its fertility potential in vitro and in vivo following fixed-time artificial insemination(Elsevier, 2020) Yang, Steve X.; Adams, Gregg P.; Palomino Cano, Jesús Manuel; Huanca, Willian F.; Lessard, Carl; Rajapaksha, Kosala; Anzar, MuhammadSuccessful cryopreservation of bison semen is fundamental for restoration of genetic diversity in Canada’s wood bison. Conventional bovine semen extenders contain animal products, such as egg yolk and milk, which are undesirable because of biosecurity risks and undefined composition. In this study, we examined the efficacy of an exogenous protein-free extender containing cholesterol-cyclodextrin complex (CC) to cryopreserve bison semen. The study also provided an opportunity to determine the effectiveness of different ovulation synchronization protocols for fixed-time artificial insemination in bison. Semen was collected from wood bison bulls via electroejaculation and cryopreserved in either Tris-egg yolk-glycerol (called ‘TEYG’) extender or pretreated with CC (2 mg/mL semen) and diluted in Tris-glycerol (collectively called ‘CC-TG’) extender. Post-thaw sperm motion characteristics and in vitro fertilization of cattle oocytes confirmed that CC alone without egg yolk protected bison sperm during cryopreservation process. In the first fertility trial, however, no pregnancy was obtained following fixed-time artificial insemination of bison cows with CC-TG extender. In a follow-up trial, low concentration of CC (1 mg/mL semen) resulted in better post-thaw sperm motion characteristics, fertility rate, and birth of live calves following fixed-time artificial insemination. Results showed that 1 mg CC/mL semen completely replaced egg yolk in bison semen extender. In addition, both follicular ablation and steroid treatment protocols provided ovulation synchrony to permit successful application of fixed-time artificial insemination in bison. This is the first report on the birth of live bison calves following fixed-time artificial insemination using semen cryopreserved in a defined extender.Ítem Solo Metadatos Effect of extending FSH treatment on superovulation and embryo collection in wood bison (Bison bison athabascae)(Elsevier, 2017) Palomino Cano, Jesús Manuel; Cervantes, Miriam P.; Mapletoft, Reuben J.; Woodbury, Murray R.; Adams, Gregg P.The effect of extending the length of the FSH treatment protocol on superovulatory response and embryo production was investigated in wood bison during the anovulatory and ovulatory seasons. In Experiment 1 (anovulatory season), follicular wave emergence was synchronized by follicular ablation (Day -1) and bison were assigned randomly to two groups (n = 14/group) and given 200 mg FSH on Day 0 and Day 2 (non-extended group), or 133 mg FSH on Days 0, 2, and 4 (extended group). Human chorionic gonadotropin (hCG; 3000 IU) was given on Day 5 and Day 6 in the non-extended and extended groups, respectively, and bison were inseminated 12 and 24 h later. Ova/embryos were collected 8 days after hCG treatment. In Experiment 2 (ovulatory season), bison were synchronized and superstimulated as in Experiment 1 (n = 12/group), but prostaglandin was given to control CL development. Data were compared by t-test and Chi-square test. In Experiment 1, no differences in ovarian response or embryo production between groups were detected. In Experiment 2, there was no difference in the ovarian response between groups, however, a greater number of ova/embryos (4.3 ± 0.8 vs. 2.3 ± 0.4; P ≤ 0.05) and freezable embryos (2.5 ± 0.6 vs. 1.2 ± 0.4; P ≤ 0.05) were obtained in the extended group. The number of freezable embryos was greater during the ovulatory vs anovulatory season (1.8 ± 0.4 vs. 0.3 ± 0.2; P ≤ 0.05). In conclusion, extending the FSH treatment in wood bison did not improve the superovulatory response during the anovulatory season, but resulted in twice as many freezable embryos during the ovulatory season. The number of freezable embryos collected during the anovulatory season was <20% that of the ovulatory season.Ítem Acceso Abierto Effects of eCG and progesterone on superovulation and embryo production in wood bison (Bison bison athabascae)(Elsevier, 2017) Palomino Cano, Jesús Manuel; Cervantes, Miriam P.; Woodbury, Murray R.; Mapletoft, Reuben J.; Adams, Gregg P.Experiments were done to determine if inclusion of eCG and progesterone in the superstimulation protocol will increase the ovarian response and embryo production in wood bison, and to provide preliminary information regarding the effect of season. In Experiment 1 (anovulatory season), bison (n = 26) were synchronized by follicular ablation (Day −1) and given FSH on Days 0 and 2, and assigned to 3 groups: Progesterone (Days 0–4), eCG (Day 3), or progesterone + eCG. On Day 5, bison were given hCG and inseminated 12 and 24 h later. Ova/embryos were collected 8 days after hCG. In Experiment 2 (ovulatory season), bison (n = 24) were synchronized and assigned randomly to two groups in which superstimulation was induced with FSH, either with or without eCG, as in Experiment 1. No differences among groups were found in ovarian response or embryo production in either experiment. The follicular count at wave emergence was positively correlated with the number of large follicles at the end of superstimulation in all groups. A significantly greater number of follicles present at wave emergence in the anovulatory vs. ovulatory season was associated with a greater number of CL at the time of embryo collection, but only half the number of freezable embryos. In conclusion, the number of transferable embryos collected (1–2/bison) was higher than in any previous report, but was not attributable to the inclusion of eCG or progesterone in the superovulatory protocol. The apparent effect of season on oocyte competence, and not superovulatory response, is worthy of further investigation.Ítem Solo Metadatos In vitro embryo production in wood bison (Bison bison athabascae) using in vivo matured cumulus-oocyte complexes(Elsevier, 2017) Cervantes, Miriam P.; Palomino Cano, Jesús Manuel; Anzar, Muhammad; Mapletoft, Reuben J.; Mastromonaco, Gabriela F.; Adams, Gregg P.Experiments were conducted in wood bison to determine the effect of additional maturation time on embryo development of in vivo matured oocytes. In experiment 1, cumulus-oocyte complexes (COC) were collected 30 hours after hCG treatment in superstimulated wood bison, and expanded COC were fertilized immediately or after 4 hours of additional in vitro maturation. Embryo development was assessed on Days 3, 7, and 8 (Day 0 = day of fertilization). No difference in cleavage rate was detected (55.3% vs. 60.5%, P = 0.82), but the Day 8 blastocyst rate was higher after an additional 4 hours of in vitro maturation time (44.7 vs. 18.4%, P = 0.03). In experiment 2, COC were collected at either 30 hours or 34 hours after hCG treatment. Expanded COC from the 30 hours group were fertilized after 4 hours of in vitro maturation, whereas those from the 34 hours group were fertilized immediately. A higher cleavage rate (74.3 vs. 57.0%) and blastocyst rate (54.1 vs. 37.2%) were found in the 34 hours group versus the 30 hours group (P < 0.05). In conclusion, an additional short period of in vitro maturation, or an extended period of in vivo maturation are beneficial for in vitro embryo production in wood bison.Ítem Solo Metadatos In vitro-production of embryos using immature oocytes collected transvaginally from superstimulated wood bison (Bison bison athabascae)(Elsevier, 2017) Cervantes, Miriam P.; Palomino Cano, Jesús Manuel; Anzar, Muhammad; Mapletoft, Reuben J.; Mastromonaco, Gabriela F.; Adams, Gregg P.Two experiments were done to test the hypothesis that morphologic characteristics of wood bison cumulus-oocyte complexes (COC) are reflective of the ability of the oocyte to develop to an advanced embryonic stage after in vitro maturation, fertilization and culture, and to determine the effect of prolonging the interval from the end of superstimulation treatment to oocyte collection (FSH starvation period). Experiments were done during the anovulatory season. In Experiment 1, ovarian superstimulation was induced in 10 bison with two doses of FSH given at 48 h intervals beginning at the time of follicular wave emergence. COC were collected 3 days (72 h) after the last dose of FSH by follicular aspiration and classified as compact, expanded or denuded. The COC were matured in vitro for 24 h before fertilization in vitro (Day 0). Embryo development was assessed on Days 3, 7 and 8. The blastocyst rate was 7/34, 2/10 and 0/3 in COC classified as compact, expanded and denuded, respectively; however, only compact COC resulted in embryos that reached the expanded blastocyst stage. In Experiment 2, COC were collected at either 3 or 4 days (72 or 96 h) after the last dose of FSH (n = 16 bison/group) to determine the effect of the duration of FSH starvation on oocyte competence. The COC were classified as compact good (>3 layers of cumulus cells), compact regular (1–3 layers of cumulus cells), expanded or denuded, and then matured, fertilized and cultured in vitro. Although follicles were larger (P < 0.05) in the 4-day FSH starvation group, there was no effect of starvation period on the distribution of COC morphology; overall, 112/194 (57.7%) were compact, 29/194 (26.3%) were expanded, 39/194 (20.1%) were denuded, and 14/194 (7.2%) were degenerated (P < 0.05). Similarly, there was no effect of starvation period on embryo development. Compact good COC had the highest cleavage (88%) and blastocyst rates (54%; P < 0.05), followed by compact regular COC at 73% and 25%, respectively. Expanded and denuded COC had low cleavage (40% vs. 59%, respectively) and blastocyst rates (5% vs. 8%, respectively). We conclude that morphologic characteristics of wood bison COC are reflective of the ability of the oocyte to develop into an embryo in vitro. Importantly, oocytes collected from superstimulated bison during the anovulatory season were competent to develop to the blastocyst stage following in vitro maturation, fertilization and culture.Ítem Solo Metadatos In vivo and in vitro maturation of oocytes collected from superstimulated wood bison (Bison bison athabascae) during the anovulatory and ovulatory seasons(Elsevier, 2016) Cervantes, Miriam P.; Palomino Cano, Jesús Manuel; Anzar, Muhammad; Mapletoft, Reuben J.; Adams, Gregg P.Experiments were done to compare the in vivo and in vitro maturational characteristics of cumulus-oocyte complexes (COC) collected from live wood bison. In Experiment 1 (anovulatory season), follicular ablation was done to synchronize follicle wave emergence among bison on Day −1, and FSH was given on Days 0 and 2. Bison were then assigned to 5 groups (n = 5/group) in which COC were collected by transvaginal follicle aspiration on Day 4 and either fixed immediately with no maturation (control), matured in vitro for 24 or 30 h, or collected on Day 5 after in vivo maturation for 24 or 30 h (i.e., after hCG treatment). In Experiment 2 (ovulatory season), bison were treated as described for Experiment 1, but PGF2α (cloprostenol) was given to control the luteal phase on Days −9 and 3. In both experiments, cumulus cell expansion was more extensive following in vivo than in vitro maturation, and the percentage of fully expanded COC was highest in the in vivo 30 h groups. Nuclear maturation occurred more rapidly in vitro; 60–70% of oocytes were at the MII stage 24 h after in vitro maturation while only 25–27% of oocytes had reached the MII stage after 24 h of in vivo maturation. In conclusion, nuclear maturation occurred more rapidly during in vitro vs. in vivo maturation, but was associated with less cumulus expansion than in vivo maturation. In vivo oocyte maturation was more complete at 30 vs. 24 h after hCG treatment. Season had no effect on the maturational capacity of wood bison oocytes.Ítem Solo Metadatos In vivo imaging in the rabbit as a model for the study of ovulation-inducing factors(Sage publications, 2014) Cervantes, Miriam P.; Palomino Cano, Jesús Manuel; Adams, Gregg P.The study of factors responsible for eliciting ovulation in rabbits has been hampered by the lack of a suitable method of monitoring the ovaries in vivo. Ovarian imaging by ultrasound biomicroscopy was used in two experiments designed to determine the effects of seminal plasma on the ovulatory response in rabbits. In Experiment 1, female rabbits were group-housed and treated intramuscularly with saline, gonadotropin releasing hormone (GnRH), or seminal plasma of llamas or rabbits (n = 4 to 6 per group). Rabbits were euthanized eight days later to evaluate the ovarian response by ultrasound biomicroscopy ex situ. No differences among groups were detected in the proportion of rabbits that ovulated or in the number and size of corpora lutea. The high incidence of ovulation in the negative control group was unexpected, and confounded determination of an ovulation-inducing effect of seminal plasma. In Experiment 2, female rabbits were caged individually, and treated as in Experiment 1 (n = 5 to 7 per group). The ovarian response was evaluated in vivo by transcutaneous ultrasound biomicroscopy. Ovulation and formation of corpora lutea were detected only in rabbits given GnRH. A preovulatory surge in plasma luteinizing hormone concentration and a post-ovulatory rise in plasma progesterone concentration were detected only in rabbits treated with GnRH. Surgical translocation of the ovaries to a subcutaneous position enabled longitudinal assessment of the ovulatory response by ultrasound biomicroscopy. Results clearly documented the effect of physical/social interaction on ovulation in rabbits, and did not support the hypothesis that seminal plasma elicits ovulation in rabbits.Ítem Acceso Abierto Inducing ovulation in wood bison (Bison bison athabascae) during the anovulatory season(Elsevier, 2015) Palomino Cano, Jesús Manuel; Cervantes, Miriam P.; Adams, Gregg P.As part of the development of a germplasm biobank to preserve the genetic diversity of threatened wood bison (Bison bison athabascae), a 2 × 2 factorial study was designed to determine the effects of ovulation induction agent and follicle maturity on the ovulatory response in wood bison during the anovulatory season. Bison (n = 32) were assigned randomly to four groups (n = 8/group) and treated with either pLH or hCG when a growing dominant follicle was either 8–9 mm or ≥10 mm. The ovaries were examined daily by ultrasonography to determine the timing of ovulation, and 7 days post-treatment to assess CL development. The proportion of bison that ovulated was greater in bison treated with hCG than pLH ([15/16] 94% vs. [8/16] 50%; P < 0.05), and when the dominant follicle was ≥10 mm vs. 8–9 mm at the time of treatment (88% vs. 56%; P < 0.05). The interval from treatment to ovulation was 37.0 ± 1.3 h and was not affected by induction agent or follicle size. However, synchrony of ovulation tended to be greater (P = 0.10) in the ≥10 mm group vs. the 8–9 mm group, and the ensuing corpus luteum was larger (15.3 ± 0.43 mm vs. 13.4 ± 0.36; P < 0.05). In conclusion, both ovulation inducing agent and follicle size influenced the ovulatory response in bison during the anovulatory season. Treatment with hCG was more effective than pLH for inducing ovulation in wood bison, and the effect was greater when treatment was given when the growing dominant follicle was ≥10 mm.Ítem Acceso Abierto Ovarian superstimulation and oocyte collection in wood bison (Bison bison athabascae) during the ovulatory season(Elsevier, 2014) Palomino Cano, Jesús Manuel; McCorkell, Robert B.; Woodbury, Murray R.; Cervantes, Miriam P.; Adams, Gregg P.The objective of the study was to establish an effective ovarian superstimulatory protocol and subsequently obtain oocytes from bison by transvaginal ultrasound-guided follicular aspiration. Two experiments involving 22 wood bison were done during the breeding season (September to December). In experiment 1, the bison were given a luteolytic dose of prostaglandin (Day 0) and underwent follicular ablation (Day 8) to induce ovarian synchrony. Synchronized bison were then assigned randomly to two groups (n = 11 per group) and given either 200 mg FSH diluted in saline sc, or 200 mg FSH diluted in a proprietary slow-release formulation (SRF) im on Days 9 and 11. Prostaglandin was given to both groups on Day 11 followed by 25 mg LH on Day 13. Oocytes were collected by transvaginal ultrasound-guided aspiration of follicles ≥5 mm on Day 14. In experiment 2, bison were synchronized as in experiment 1 and assigned randomly to one of two groups (n = 11 per group) and given either a single dose of 2500 IU eCG im on Day 9, or 200 mg FSH sc on Days 9 and 11. Prostaglandin was given to both groups on Day 11, and LH (25 mg) was given on Day 13. Oocyte collection was done as described in experiment 1. Cumulus-oocyte-complexes (COC) were classified according to morphologic characteristics. In experiment 1, more follicles ≥5 mm were detected on Day 14 in bison treated with FSH versus eCG (12.2 ± 1.73 vs. 5.8 ± 0.52; P < 0.05), and more COC were collected from FSH-treated animals (7.2 ± 1.41 vs. 3.4 ± 0.62; P < 0.05). In experiment 2, the FSH-saline and FSH-SRF groups had a similar number (mean value ± standard error of the mean) of follicles ≥5 mm on Day 14 (12.4 ± 1.49 vs. 13.8 ± 1.24, respectively) and a similar number of COC were collected (6.5 ± 1.13 vs. 6.3 ± 0.96, respectively). The proportion of COC collected per follicle aspirated and the percentage of compact, expanded, and denuded oocytes did not differ between groups in either experiment 1 or 2. In summary, a two-dose regimen of FSH diluted in saline and given sc or in a SRF and given im induced a similar ovarian response in wood bison, whereas a single dose of eCG resulted in a significantly lower ovarian response. Overall, COC were collected from 55% of follicles after transvaginal, ultrasound-guided needle aspiration in wood bison.Ítem Solo Metadatos Ovarian synchronisation in wood bison (Bison bison athabascae) during the anovulatory season(Commonwealth Scientific and Industrial Research Organisation, 2013) Palomino Cano, Jesús Manuel; McCorkell, Robert B.; Woodbury, Murray R.; Adams, Gregg P.Two experiments were performed in wood bison during the anovulatory season to establish an effective protocol for ovarian synchronisation. In an untreated control phase, bison cows (n = 19) were examined daily to establish the interval to new follicular wave emergence (4.9 ± 0.7 days) for the purposes of comparison with the experimental treatments. In Experiment 1, bison were treated by transvaginal ultrasound-guided follicular ablation (n = 9) or with 2 mg, i.m., 17β-oestradiol (n = 10). In Experiment 2, bison were treated by follicular ablation (n = 9) or with 2 mg, i.m., 17β-oestradiol +100 mg, i.m., progesterone (n = 10). In Experiment 1, the interval to new wave emergence for control, follicular ablation and 17β-oestradiol-treated groups was 4.9 ± 0.7, 1.1 ± 0.1 and 3.1 ± 0.4 days, respectively (P < 0.05). The degree of synchrony was 2.4 ± 0.4, 0.2 ± 0.1 and 0.8 ± 0.2 days, respectively (P < 0.05). In Experiment 2, the interval to new wave emergence for control, follicular ablation and 17β-oestradiol + progesterone-treated groups was 4.9 ± 0.7, 1.2 ± 0.2 and 3.3 ± 0.3 days, respectively (P < 0.05), and the degree of synchrony was 2.4 ± 0.4, 0.2 ± 0.1, and 0.8 ± 0.2 days, respectively (P < 0.05). The degree of synchrony did not differ between ablation and hormone treatment groups in either experiment, but was greater in treatment groups than in the untreated control phase. Both follicular ablation and hormone treatment shortened and decreased the variability in the interval to follicular wave emergence in bison, but wave emergence occurred earlier after follicular ablation.Ítem Acceso Abierto Superstimulatory response and oocyte collection in North American bison during the non-breeding season(Elsevier, 2013) Palomino Cano, Jesús Manuel; McCorkell, Robert B.; Woodbury, Murray R.; Cervantes, Miriam P.; Adams, Gregg P.A 2 × 2 design was used to compare the ovarian response and oocyte collection characteristics in bison given a superstimulatory dose of eCG or FSH, with or without a follow-up dose of LH. Follicular wave emergence was synchronized by follicle ablation (Day −1) and bison were assigned randomly to two superstimulatory treatment groups (n = 10 per group): (i) a single intramuscular dose of 2500 IU of eCG given on Day 0, or (ii) two subcutaneous doses of 200 mg of FSH given on Days 0 and 2. On Day 4, 200 mg of LH was given intramuscularly in 5 bison in each superstimulatory treatment group. The study was done in two replicates (n = 20 per replicate) involving a crossover design so that each animal was given the opposite superstimulatory treatment (eCG or FSH) during successive replicates. Cumulus-oocyte complexes (COC) were collected by transvaginal ultrasound-guided follicle aspiration, and were classified according to morphologic attributes as compact, expanded, or denuded. Superstimulatory treatment with FSH (vs. eCG) resulted in the development of more follicles ≥5 mm (14.2 ± 1.41 vs. 8.2 ± 0.67; P < 0.05; mean ± SEM), and more follicles aspirated (12.4 ± 1.3 vs. 6.3 ± 0.6; P < 0.04). Follow-up treatment with LH (vs. no LH) resulted in a greater proportion of expanded COC (37% vs. 15%; P < 0.05), and a tendency for a higher COC collection rate (61% vs. 54%; P = 0.08). In summary, superstimulation with FSH (vs. eCG) resulted in twice as many follicles available for aspiration and nearly twice as many COC collected in bison during the anovulatory season, and follow-up treatment with LH increased the proportion of expanded COC collected.