Journal articles on the topic 'Superstimulation treatment'
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Rivera, Fernando A., Luís G. D. Mendonça, Gláucio Lopes, José E. P. Santos, Rolando V. Perez, Marcel Amstalden, Abelardo Correa-Calderón, and Ricardo C. Chebel. "Reduced progesterone concentration during growth of the first follicular wave affects embryo quality but has no effect on embryo survival post transfer in lactating dairy cows." REPRODUCTION 141, no. 3 (March 2011): 333–42. http://dx.doi.org/10.1530/rep-10-0375.
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Fertility of lactating dairy cows is associated with reduced progesterone (P4) concentration compared with nonlactating animals. The objective of the current study was to determine whether P4 during growth of the first follicular wave (FFW) affects embryo quality. Lactating Holstein cows at 33±3 days post partum were allocated to one of three treatments. Cows in the FFW and FFW with P4 (FFWP) treatments started the superstimulation protocol on day 1 of the estrous cycle and second follicular wave (SFW) cows started the superstimulation protocol on estrous cycle day 7. Cows were superstimulated with 400 mg of NIH-FSH-P1 (FSH) given twice daily for 5 days, two prostaglandin F2α (PGF2α) injections given with the ninth and tenth injections of FSH, GNRH given 48 h after the first PGF2α injection, and timed insemination 12 and 24 h after the GNRH injection. Cows in the FFWP treatment received two intravaginal P4 inserts during the superstimulation. Embryos were recovered 6.5 days after artificial insemination and excellent/good and fair embryos were frozen and transferred. Blood was sampled daily from estrous cycle day 0 until insemination from donor cows. During the superstimulation protocol, P4 was (P<0.01) greatest for SFW cows followed by FFWP and FFW cows respectively. The percentage of embryos–oocytes from SFW and FFWP cows classified as excellent/good and fair embryos was (P=0.02) greater than those of FFW cows. Pregnancy per embryo transfer was not (P≥0.73) affected by embryo donor treatment. Reduced embryo quality of cows induced to ovulate the follicles from the first follicular wave is a consequence of reduced P4 during follicle growth.
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Carballo Guerrero, D., A. Tribulo, R. Tribulo, H. Tribulo, and G. A. Bo. "294 SUPERSTIMULATION IN THE FIRST FOLLICULAR WAVE, WITHOUT THE USE OF ESTRADIOL IN BONSMARA CATTLE." Reproduction, Fertility and Development 20, no. 1 (2008): 226. http://dx.doi.org/10.1071/rdv20n1ab294.
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Protocols that control follicular wave emergence and ovulation have had a great impact on the application of commercial on-farm embryo transfer because they permit the initiation of superstimulatory treatments at a self-appointed time. However, the most common approach for the synchronization of follicular wave emergence for superstimulation involves the use of estradiol or its esters that are not commercially available in many countries. Therefore, an experiment was designed to evaluate a protocol in which the superstimulation treatment began at the time of emergence of the first follicular wave without the use of estradiol. Bonsmara donors (29 cows and 41 heifers) were randomly allocated to one of two treatment groups. Donor animals in the experimental group (first wave group) received an intravaginal progesterone releasing device (Cue-Mate, Bioniche Animal Health, ON, Canada) along with PGF (0.150 mg D + cloprostenol, Bioprost-D, Biotay, Argentina) at random stages of the estrous cycle. Cue-Mates were removed 10.5 d later and a second PGF was administered at the same time, followed by GnRH (0.050 mg Lecirelina, Biosin-OV, Biotay, Argentina) 36 h later. Ovulation was expected to occur within 30 h after GnRH (day 0). On day 0 (36 h after gonadotropin-releasing hormone) donors received a new Cue-Mate, and superstimulation treatment was initiated with a total dose of 200 to 260 mg (heifers) or 320 mg (cows) NIH-FSH-P1of Folltropin-V in twice daily decreasing doses over 5 d. The PGF was administered with the last two Folltropin-V injections, and Cue-Mate devices were removed with the last Folltropin-V injection. All donors received 12.5 mg pLH (Lutropin-V, Bioniche Animal Health) 24 h after Cue-Mate removal and were AI 12 and 24 h later. Embryos were collected 7 d after pLH treatment. Donors in the Control group received a Cue-Mate and 2 mg of estradiol benzoate (EB; Bioestradiol, Biotay) and 50 mg of progesterone (Lab. Rio de Janeiro, Argentina), and superstimulation treatments were initiated 4 d later with the same dosages used in the first wave group. The PGF administration, Cue-Mate removal, AI, and embryo collections were done as those in the first wave group. Data were analyzed by ANOVA, and results are shown in Table 1. It was not possible to pass the cervix with the collection catheter in two heifers in the control group, and they were excluded from the analysis. There were no significant effects of donor category (cows v. heifers) or treatment on superovulatory response and embryo quality (P > 0.20). In conlusion, superstimulation on a synchronized first follicular wave is as efficacious as superstimulation following synchronization of follicle wave emergence with estradiol benzoate in Bonsmara cattle. Table 1. Superovulatory response (means ± SEM) in Bonsmara cows and heifers treated with Folltropin-V during the first follicular wave or 4 d after estradiol administration Bioniche Animal Health, Belleville, ON, Canada.
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Carballo Guerrero, D., A. Tríbulo, R. Tríbulo, H. Tríbulo, and G. A. Bó. "404 SUPEROVULATORY RESPONSE IN BEEF DONORS TREATED DURING THE FIRST FOLLICULAR WAVE OR FOUR DAYS AFTER PROGESTERONE AND ESTRADIOL ADMINISTRATION." Reproduction, Fertility and Development 22, no. 1 (2010): 358. http://dx.doi.org/10.1071/rdv22n1ab404.
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Although we have previously shown that ovarian superstimulation during the first follicular wave resulted in a successful response (Carballo Guerrero D et al. 2009 Reprod. Fertil. 21, 242), the current protocol needs to be optimized in order to be used in the field. Therefore, an experiment was designed to simplify this treatment and to compare it with the traditional superstimulation protocol using progesterone and estradiol. Simmental cows (n = 14) were subjected to 3 superstimulation treatments (2 first wave groups and 1 control group) in a crossover design (i.e. all cows received the 3 treatments and all treatments were represented on each collection day). Cows in Group 1 received a progesterone-releasing device (Cue-Mate®, Bioniche Animal Health, Belleville, Ontario, Canada) along with 0.150 mg of D + cloprostenol (PGF; Bioprost-D®, Biotay, Buenos Aires, Argentina) at random stages of the estrous cycle. A second PGF was injected 5 days after Cue-Mate® insertion, followed by GnRH (0.050 mg of lecirelin; Biosin-OV®, Biotay) 36 h later (i.e. 7 days after Cue-Mate® insertion). Based on previous studies, ovulation was expected to occur 30 to 36 h later. Therefore, superstimulation treatments were initiated 36 h after GnRH (Day 0), with a total dose of 400 mg NIH-FSH-P1 of Folltropin®-V (Bioniche Animal Health) in twice-daily decreasing doses over 4 days. Prostaglandin was administered with the last 2 Folltropin®-V injections and Cue-Mate® devices were removed with the last Folltropin®-V injection. Cows received 12.5 mg of porcine LH (Lutropin®-V, Bioniche Animal Health) 24 h after Cue-Mate® removal and were AI 12 and 24 h later. Ova/embryos were collected 7 days after porcine LH and evaluated following IETS recommendations. Cows in Group 2 were treated similarly to those in the Group 1, except they did not receive the second PGF injection 5 days after Cue-Mate® insertion (thus eliminating the need to handle animals on that day). Finally, cows in Group 3 [estradiol benzoate (EB)+P4 control group] received a Cue-Mate® plus 2.5 mg of EB (Bioestradiol®, Biotay) and 50 mg of progesterone (P4; Lab., Rio de Janeiro, Argentina) at random stages of their estrous cycle. Superstimulation treatments were initiated 4 days later (Day 0) following the same protocol used in Group 1. Data were transformed to square root and analyzed by ANOVA. Mean (± SEM) numbers of ova/embryos collected, fertilized ova, and transferable embryos did not differ among groups (12.9 ± 2.0, 9.8 ± 1.7, and 6.6 ± 1.2; 11.5 ± 1.7, 9.3 ± 1.5, and 7.7 ± 1.6; and 14.5 ± 2.8, 9.4 ± 2.3, and 6.8 ± 1.7 for Groups 1, 2, and 3, respectively). In conclusion, data demonstrated that superstimulation during the first follicular wave can be successfully used in groups of randomly cycling donors without the need for estrus detection or estradiol to synchronize follicular wave emergence. The protocol is easy to follow and embryo production is comparable to that of the estradiol and progesterone protocol.
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Carballo Guerrero, D., A. Tríbulo, R. Tríbulo, H. Tríbulo, and G. A. Bó. "291 SUPEROVULATORY RESPONSE IN BEEF CATTLE TREATED DURING THE FIRST FOLLICULAR WAVE FOLLOWING SYNCHRONIZATION OF OVULATION WITH A PROGESTIN DEVICE AND GnRH." Reproduction, Fertility and Development 21, no. 1 (2009): 242. http://dx.doi.org/10.1071/rdv21n1ab291.
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Although we have previously shown that superstimulation during the first follicular wave resulted in a successful response (Carballo Guerrero D et al. 2007 Reprod. Fertil. Dev. 20, 226), the protocol required many interventions that could influence its application in the field. Therefore, two studies were designed to simplify the superstimulation treatment protocol. Experiment 1 was designed to determine whether it was necessary to remove the progesterone releasing device during the superstimulation protocol. Angus cows (n = 37) were superstimulated by two treatments in a crossover design. Cows in Group 1 (control) received a progesterone releasing device (Cue-Mate, Bioniche Animal Health, Belleville, ON, Canada) along with 0.150 mg D cloprostenol (PGF, Bioprost-D, Biotay, Argentina) IM, at random stages of the estrous cycle. Five days later, a second PGF was injected and Cue-Mates were removed, followed by GnRH (0.050 mg Lecirelina, Biosin-OV, Biotay) 36 h later; ovulation was expected to occur 30 to 36 h later. On Day 0 (36 h after GnRH) donors received a new Cue-Mate and superstimulation treatment was initiated with a total dose of 400 mg NIH-FSH-P1 of Folltropin-V (FSH, Bioniche Animal Health) in twice daily decreasing doses over 5 days. PGF was injected with the last two FSH injections and Cue-Mates were removed with the last FSH injection. Cows in Group 2 were treated similarly to those in the control group, except that Cue-mate devices were not replaced and remained in place for 13 days (i.e. Cue-mates were removed with the last FSH and PGF injection). All donors received 12.5 mg pLH (Lutropin-V, Bioniche Animal Health) 24 h after Cue-Mate removal and were AI 12 and 24 h later. Embryos were collected 7 days after pLH. Means were compared between groups by Student’s t-test. Superovulatory response and embryo production did not differ between groups. Mean (± SEM) number of ova/embryos collected and transferable embryos were 8.2 ± 1.0 and 4.1 ± 0.6 v. 9.8 ± 0.9 and 5.7 ± 0.7 for Groups 1 and 2, respectively (P > 0.2). Experiment 2 was designed to evaluate the effect of giving FSH for 4 v. 5 days. Simmental (n = 18) and Angus (n = 6) cows were superstimulated by the two treatment protocols in a crossover design. Cows in both groups were treated similarly to those in Group 2 in Experiment 1 (i.e. Cue-Mates were not replaced during treatment). Cows in Group 1 (control) received FSH over 5 days (as in Group 2 of Experiment 1); while those in Group 2 received the same dosage of FSH, but given in twice daily decreasing doses over 4 days (Cue-Mates were removed with the last FSH and PGF injections). Superovulatory response and embryo production did not differ among groups. Mean (± SEM) number of ova/embryos collected and transferable embryos were 13.5 ± 2.4 and 6.6 ± 1.1 v. 12.0 ± 1.9 and 5.8 ± 1.0 for Groups 1 and 2, respectively (P > 0.6). In conclusion, superstimulation of cattle at the time of emergence of the first follicular wave after ovulation results in an acceptable superovulatory response and all treatments evaluated were user-friendly and equally efficient.
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Aboulghar, Mohamed A., Ragaa T. Mansour, Gamal I. Serour, Yehia Amin, Amal M. Abbas, and Iman M. Salah. "Ovarian superstimulation and intrauterine insemination for the treatment of unexplained infertility." Fertility and Sterility 60, no. 2 (August 1993): 303–6. http://dx.doi.org/10.1016/s0015-0282(16)56102-0.
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Dias, F. C. F., M. I. R. Khan, M. A. Sirard, G. P. Adams, and J. Singh. "Differential gene expression of granulosa cells after ovarian superstimulation in beef cattle." REPRODUCTION 146, no. 2 (August 2013): 181–91. http://dx.doi.org/10.1530/rep-13-0114.
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Microarray analysis was used to compare the gene expression of granulosa cells from dominant follicles with that of those after superstimulatory treatment. Cows were allocated randomly to two groups (superstimulation and control, n=6/group). A new follicular wave was induced by ablation of follicles ≥5 mm in diameter, and a progesterone-releasing device controlled internal drug release (CIDR) was placed in the vagina. The superstimulation group was given eight doses of 25 mg FSH at 12-h intervals starting from the day of wave emergence (day 0), whereas the control group was not given FSH treatment. Both groups were given prostaglandin F2α twice, 12 h apart, on day 3 and the CIDR was removed at the second injection; 25 mg porcine luteinizing hormone (pLH) was given 24 h after CIDR removal, and cows were ovariectomized 24 h later. Granulosa cells were collected for RNA extraction, amplification, and microarray hybridization. A total of 190 genes were downregulated and 280 genes were upregulated. To validate the microarray results, five genes were selected for real-time PCR (NTS, FOS, THBS1, FN1, and IGF2). Expression of four genes increased significantly in the three different animals tested (NTS, FOS, THBS1, and FN1). The upregulated genes are related to matrix remodeling (i.e. tissue proliferation), disturbance of angiogenesis, apoptosis, and oxidative stress response. We conclude that superstimulation treatment i) results in granulosa cells that lag behind in maturation and differentiation (most of the upregulated genes are markers of the follicular growth stage), ii) activates genes involved with the NFE2L2 oxidative stress response and endoplasmic reticulum stress response, and iii) disturbs angiogenesis.
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Barros, Ciro M., Rafael A. Satrapa, Anthony C. S. Castilho, Patrícia K. Fontes, Eduardo M. Razza, Ronaldo L. Ereno, and Marcelo F. G. Nogueira. "Effect of superstimulatory treatments on the expression of genes related to ovulatory capacity, oocyte competence and embryo development in cattle." Reproduction, Fertility and Development 25, no. 1 (2013): 17. http://dx.doi.org/10.1071/rd12271.
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Multiple ovulation (superovulation) and embryo transfer has been used extensively in cattle. In the past decade, superstimulatory treatment protocols that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-time AI (FTAI), have been developed for zebu (Bos indicus) and European (Bos taurus) breeds of cattle. There is evidence that additional stimulus with LH (through the administration of exogenous LH or equine chorionic gonadotrophin (eCG)) on the last day of the superstimulatory treatment protocol, called the ‘P-36 protocol’ for FTAI, can increase embryo yield compared with conventional protocols that are based on the detection of oestrus. However, inconsistent results with the use of hormones that stimulate LH receptors (LHR) have prompted further studies on the roles of LH and its receptors in ovulatory capacity (acquisition of LHR in granulosa cells), oocyte competence and embryo quality in superstimulated cattle. Recent experiments have shown that superstimulation with FSH increases mRNA expression of LHR and angiotensin AT2 receptors in granulosa cells of follicles >8 mm in diameter. In addition, FSH decreases mRNA expression of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in oocytes, but increases the expression of both in cumulus cells, without diminishing the capacity of cumulus–oocyte complexes to generate blastocysts. Although these results indicate that superstimulation with FSH is not detrimental to oocyte competence, supplementary studies are warranted to investigate the effects of superstimulation on embryo quality and viability. In addition, experiments comparing the cellular and/or molecular effects of adding eCG to the P-36 treatment protocol are being conducted to elucidate the effects of superstimulatory protocols on the yield of viable embryos.
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Čech, S., V. Havlíček, M. Lopatářová, M. Vyskočil, and R. Doležel. "Effects of superstimulation with fsh on follicular population and recovery rate of oocytes in the growing phase of the first and second follicular wave." Veterinární Medicína 47, No. 2 - 3 (March 30, 2012): 33–37. http://dx.doi.org/10.17221/5800-vetmed.
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Effectiveness of in vitro production of embryos (IVP) is limited among other factors by the recovery rate of oocytes. Gonadotropin superstimulation can improve the recovery rate of oocytes. The effect of FSH treatment on follicular population and recovery rate of oocytes at ovum pick-up (OPU) in the growing phase of the 1st as well as the 2nd follicular wave after superstimulation was the object of our experiment. Twelve unpregnant milking cows (15–20 kg milk per day) housed on a dairy farm were used in the experiment. The cows bearing corpus luteum were synchronized by PGF<sub>2 </sub>(day 0) and they were treated by FSH (Folicotropin inj. sicc. ad us vet., Spofa Prague, Czech Republic, single doses 80, 80, 80, 80, 40 and 40 UI) in 12 h intervals on days 12, 13 and 14. Transvaginal ultrasonographic puncture of oocytes in cows bearing a new corpus luteum was performed on day 7 (OPU 1, various phase of the follicular wave, removal of the dominant follicle) and it was repeated on days 10 (OPU 2, growing phase of the follicular wave – control), 16 (OPU 3, growing phase of the first follicular wave after superstimulation) and 20 (OPU 4, growing phase of the second follicular wave after superstimulation). All follicles > 2 mm were punctured. The ovarian follicles (ultrasonographically) and numbers and qualities of obtained oocytes (microscopically) were evaluated during and immediately after OPU. Follicular population was divided to small (FS, 2–5 mm), medium (FM, 5–9 mm) and large (FL, > 9 mm) follicles. Oocytes were classified as 1st (intact cumulus, > 3 layers of cumulus cells), 2nd (complete 1–3 layers of cumulus cells), 3rd (incomplete layers of cumulus cells, expanded cumulus mass) and 4th (absence of corona cells, degenerated oocytes) classes. Although we found the least of FS (x = 1.0) during OPU 3, significantly more FM (x = 24.7) and FL (x = 3.1) follicles were found at this procedure in comparison with others. Likewise a significantly higher number of oocytes (x = 8.1) was obtained at OPU 3 in comparison with OPU 1 and OPU 2. Significantly higher number of FM (x = 6.1) was found and non-significantly higher number of oocytes was obtained at OPU 4 in comparison with OPU 1 and 2. The results show that administration of FSH increases the number of follicles and the number of collected oocytes in the growing phase of the 1st follicular wave after superstimulation, nevertheless a higher number of follicles and a higher recovery rate of oocytes can be expected in the growing phase of the 2nd follicular wave after superstimulation as well.
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Vieira, L. M., G. A. Bó, and R. J. Mapletoft. "248 SUPERSTIMULATION STRATEGIES FOR OVUM PICKUP IN HOLSTEIN DONORS." Reproduction, Fertility and Development 28, no. 2 (2016): 256. http://dx.doi.org/10.1071/rdv28n2ab248.
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In vitro embryo production (IVP) is an important tool to enhance genetic gain in cattle. However, oocyte quality is a limiting factor for the success of IVP programs in high-producing donors. A series of studies using protocols for follicular wave synchronization and superstimulation before ovum pickup were performed to improve the efficiency of ovum pickup and in vitro production in dairy cattle. The first study evaluated superstimulation with FSH (Folltropin-V®) before ovum pickup in lactating (n = 15) and non-lactating (n = 15) Holstein donors in a crossover design. Cows underwent synchronization of follicle wave emergence (FWE) and at the expected time of FWE, the FSH group received a total dosage of 200 mg of FSH in 4 decreasing doses 12 h apart; controls received no FSH, and ovum pickup was conducted 72 h after the expected FWE in all cows. The FSH-treated cows had a higher (P < 0.01) percentage of medium-sized follicles (6 to 10 mm) at the time of ovum pickup (55.1%) than control cows (20.8%) as well as lower cumulus‐oocyte complexes (COC) recovery rates (60.0 v. 69.8%, respectively; P = 0.002). However, FSH-treated cows had a higher blastocyst production rate (34.5 v. 19.8%; P < 0.01) and more transferable embryos per ovum pickup session (3.0 ± 0.5 v. 1.8 ± 0.4; P = 0.02). Subsequent trials evaluated plasma FSH profiles in 23 heifers and in vitro production following ovum pickup in 90 non-lactating Holstein donors superstimulated with a single IM injection of FSH in 0.5% hyaluronan (HA; MAP-5®, 50 mg). Controls received no treatment, while the F200 group received 200 mg of FSH in 4 decreasing doses 12 h apart. The F200HA and F300HA groups received 200 or 300 mg of FSH in 5 or 7.5 mL, respectively, of 0.5% HA by a single IM injection. Circulating FSH area under curve (AUC) in FSH-treated animals was greater than in the control group (P = 0.02). Although the AUC for F200 group did not differ from HA groups (P = 0.56), the total period of time plasma FSH levels were elevated was greater than in the HA groups (P < 0.01). In the IVP trial, FSH-treated cows had a greater proportion of medium-sized (6–10 mm) follicles than controls (P < 0.001). Also, numbers of follicles (P = 0.01) retrieved (control: 13.1 ± 1.0; F200: 16.5 ± 1.2; F200HA: 19.5 ± 2.1; F300HA: 15.4 ± 1.4; P = 0.01) and blastocysts produced per ovum pickup session (control: 2.4 ± 0.5; F200: 3.7 ± 0.7; F200HA: 4.7 ± 0.7; F300HA: 3.1 ± 0.6; P = 0.06) were greater in cows receiving FSH, regardless of treatment. Cows in the F200HA group had a greater recovery rate (P = 0.009), number of COC cultured (P = 0.04), and blastocysts per ovum pickup session (P = 0.06) than cows in the F300HA group. In conclusion, superstimulation of Holstein donors before ovum pickup increased the efficiency of in vitro production. Additionally, a single IM dose of FSH in 0.5% HA resulted in similar plasma FSH profiles to twice-daily FSH treatment. Non-lactating donors treated with FSH produced more embryos per ovum pickup session regardless of FSH treatment. Lastly, all in vitro-produced endpoints were greater following a single dose of 200 mg of FSH in 0.5% HA than 300 mg of FSH in 0.5% HA.
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Bó, Gabriel A., and Reuben J. Mapletoft. "Superstimulation of ovarian follicles in cattle: Gonadotropin treatment protocols and FSH profiles." Theriogenology 150 (July 2020): 353–59. http://dx.doi.org/10.1016/j.theriogenology.2020.02.001.
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Aboulghar, Mohamed A., Ragaa T. Mansour, and Gamal I. Serour. "Ovarian superstimulation in the treatment of infertility due to peritubal and periovarian adhesions." Fertility and Sterility 51, no. 5 (May 1989): 834–37. http://dx.doi.org/10.1016/s0015-0282(16)60675-1.
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Menchaca, A., M. Vilariño, M. Crispo, T. de Castro, and E. Rubianes. "New approaches to superovulation and embryo transfer in small ruminants." Reproduction, Fertility and Development 22, no. 1 (2010): 113. http://dx.doi.org/10.1071/rd09222.
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The present paper reviews the current state of knowledge in multiple ovulation and embryo transfer (MOET) technology in small ruminants, focusing on recently reported information. Major new findings are related to follicular wave patterns in small ruminants, the elucidation of follicular dominance and the integration of this information into ovarian superstimulatory treatment protocols. Follicular dynamics determine steroid and gonadotrophin secretion, follicular responses to FSH, ovulatory responses and embryo yield. Protocols that control follicular dominance have been designed to allow the initiation of superstimulation at the beginning of a follicular wave. New approaches consist of (1) synchronisation of ovulation for superstimulation during Wave 1 (i.e. Day 0 protocol), (2) pretreatment with a gonadotrophin-releasing hormone (GnRH) antagonist from 10 days prior to FSH treatment to avoid follicular dominance and (3) progesterone–oestradiol cotreatment to synchronise follicle wave emergence. These protocols provide a homogeneous pool of small follicles that are gonadotrophin responsive, enhancing the superovulatory response and embryo yield with a reduction in the incidence of unovulated follicles and early regression of corpora lutea. In addition, the rate of fertilisation failure has been reduced by using an inducer of ovulation (i.e. GnRH) associated with intrauterine insemination. In summary, the application of recently acquired knowledge has resulted in relevant improvements in MOET programmes in small ruminants.
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Chumchai, Rujira, Thanaporn Ratsiri, Ruthaiporn Ratchamak, Wuttigrai Boonkum, and Vibuntita Chankitisakul. "Superovulatory responses based on ovarian sizes after superstimulation in Thai-Holstein crossbred dairy cows." Veterinary Integrative Sciences 20, no. 1 (September 6, 2021): 49–59. http://dx.doi.org/10.12982/vis.2022.005.
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This study was designed (1) to examine the relationship between ovary sides/sizes after superstimulation treatment and ovulatory responses in terms of large follicles, corpora lutea (CLs) numbers, and ovulation rate; and (2) to evaluate the coefficient of determination (R2) as a tool to predict the subsequent superovulatory responses by ovary sizes after superstimulation treatment in the Thai-Holstein crossbreed dairy cows. Data included 33 records from 12 superovulated Thai Holstein crossbreds. Cows were estrus synchronized on day 0 and superstimulated with 400 mg of FSH with decreasing doses twice daily for 4 days. After superovulatory treatment (day 9), the sizes of ovaries were measured and divided into 3 groups by quarters according to the ovarian sizes. Group A (< 816 mm2) ovaries were 25% smaller and group C (> 1449 mm2) ovaries were 25% larger than group B ovaries (816–1449 mm2). On day 9 and 16, there were no significant differences in the average ovary area (p > 0.05). The numbers of large follicles and CLs of group B and C were greater than those of group A (p < 0.05). The ovulation rate did not differ among groups (p > 0.05). The moderate R2 score between ovary size after superovulatory treatment and the numbers of dominant follicles and CLs were calculated (R2 = 0.445 and 0.370, p <0.05) while the beta coefficient (b-value) was positive for both observation parameters. In conclusion, the numbers of large follicles and CLs related to the size of ovaries after superovulation treatment. The moderate R2 score obtained in this study could be indicative of the limited possibility for using ovary size after superovulatory treatment for predicting superovulatory responses
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Somfai, T., K. Imai, M. Kaneda, S. Akagi, S. Haraguchi, S. Watanabe, Y. Inaba, M. Geshi, and T. Nagai. "138 THE EFFECT OF FOLLICLE SUPERSTIMULATION ON mRNA LEVELS IN BOVINE OOCYTES COLLECTED BY OVUM PICKUP." Reproduction, Fertility and Development 24, no. 1 (2012): 181. http://dx.doi.org/10.1071/rdv24n1ab138.
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A previous study revealed that follicle superstimulation significantly improved the developmental competence of immature bovine oocytes collected by ovum pickup (OPU; Imai et al. 2008 Reprod. Fertil. Dev. 20, 182). The aim of the present study was to investigate the effect of follicle superstimulation on the expression of developmentally important genes in bovine oocytes collected by OPU. Follicular oocytes were collected by OPU without (OPU group) or after follicle superstimulation by FSH (FSH/OPU group) by using a 7.5-MHz linear transducer with needle connected to an ultrasound scanner according to Imai et al. (2008). In the FSH/OPU group, after dominant follicle removal from Holstein dry cows by OPU, a CIDR was inserted on Day 5 (dominant follicle removal = Day 0). Cows then received 30 mg of FSH twice a day from Days 7 to 10 in decreasing doses (6, 6, 4, 4, 3, 3, 2, 2 mg) by IM injection. Cloprostenol (PGF; Clopromate C; Sumitomo Pharmaceuticals Co., Tokyo, Japan; 0.75 mg) was administered in the morning of Day 9 (third day of superstimulation). Oocyte collection by OPU was performed 48 h after PGF administration (Day 11) by the aspiration of follicles larger than 5 mm in diameter. In the OPU group, 3-to-6-mm follicles were aspirated without any previous hormone treatment. In vitro oocyte maturation (IVM) was performed according to Imai et al. (2006 J. Reprod. Dev. 52(Suppl), 19–29). Gene expression was assessed before (0 h IVM) and after IVM (22 h IVM) by RT quantitative PCR. The following genes were investigated: GAPDH, G6PDH, ACTB, H2A, CCNB1, MnSOD, OCT4, SOX2, CX43, HSP70, GLUT8, PAP, GDF9, COX1, ATP1A1, CDH1, CTNNB1, AQP3, DYNLL1, DYNC 1/1 and PMSB1. In brief, mRNA was extracted from 20 oocytes per sample using Qiagen RNeasy Micro kit (Qiagen, Valencia, CA, USA). Gene expression was analysed by a Roche Light Cycler 480 device. The relative expression of each gene was normalized to ACTB. Three replications were performed. Data were analysed by ANOVA. At 0 h IVM, PAP and DYNC 1/1 were found to be down-regulated (P < 0.05) in the FSH/OPU group compared with the OPU group, whereas the rest of the studied genes showed similar expression in the FSH/OPU and OPU groups. At 22 h IVM, PAP and DYNC 1/1 remained down-regulated in FSH/OPU oocytes. However, at this time the expression of GDF9 appeared significantly higher (P < 0.05) in FSH/OPU oocytes than in OPU oocytes. The expression of GDF9 was found to decrease during IVM in both groups; however, this decrease was less drastic in FSH/OPU oocytes. The results suggest that follicle superstimulation caused reduced expression of mRNA levels of PAP and DYNC 1/1 irrespective of maturation status and it also moderated the reduction of mRNA levels of GDF9 during IVM.
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Tríbulo, A., H. Tríbulo, R. Tríbulo, D. Carballo Guerrero, P. Tríbulo, D. Rogan, R. J. Mapletoft, and G. A. Bó. "420 SUPERSTIMULATION OF ANGUS DONORS WITH A SINGLE INTRAMUSCULAR INJECTION OF FOLLTROPIN®-V." Reproduction, Fertility and Development 22, no. 1 (2010): 367. http://dx.doi.org/10.1071/rdv22n1ab420.
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Two experiments were designed to determine the superovulatory response of Angus donors treated with a single injection of Folltropin®-V diluted in a slow release formulation (SRF; Bioniche Animal Health Inc., Belleville, Ontario, Canada). Experiment 1 was designed to compare superovulatory response in Angus donor cows (n = 29 per group) treated with a single i.m. injection of Folltropin®-V diluted in SRF with cows treated using the traditional twice-daily i.m. injection treatment schedule. On Day 0, all cows received 5 mg of estradiol-17β plus 50 mg of progesterone and a Cue-Mate® (Bioniche Animal Health). On Day 4, cows were superstimulated with 400 mg of NIH-FSH-P1 Folltropin®-V in twice-daily decreasing doses over 4 days or in a single i.m. injection in the neck. The single injection was prepared by diluting the Folltropin®-V lyophilized powder in 1 mL of saline for injection and mixed with 9 mL of the SRF in the syringe immediately before administration. In the am and pm of Day 6, all cows received PGF2 and Cue-Mates® were removed in the pm. In the am of Day 8, cows received 12.5 mg of porcine LH (Lutropin®-V; Bioniche Animal Health) and were inseminated 12 and 24 h later. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS recommendations. Means were compared between groups by t-test Mean (±SEM) number of ova/embryos and transferable embryos were 13.7 ± 2.1 and 7.1 ± 1.3 v. 12.3 ± 1.5 and 5.3 ± 0.8 for donors treated with the single v. twice-daily injections, respectively (P > 0.4). Experiment 2 was designed to confirm the results of Experiment 1 and to compare the effect of different dosages of Folltropin®-V on embryo production in Angus cows. Cows (n = 23) were superstimulated by 6 treatment protocols (2 × 3 factorial) in a crossover design (i.e. all cows received the 6 treatments and all treatments were represented on each collection day). Cows received the same treatments as cows in Experiment 1 except that the dosages of Folltropin®-V used were 200, 300, or 400 mg. Statistical analysis (ANOVA) revealed a significant effect of dosage of Folltropin®-V on embryo production, but there was no effect of treatment (i.e. single v. twice-daily injections; P > 0.2), nor was there a treatment by dosage interaction (P > 0.7). Mean (± SEM) number of ova/embryos and transferable embryos were 10.3 ± 0.9 and 5.4 ± 0.6 v. 11.3 ± 0.6 and 5.6 ± 0.5 for donors treated with the single or twice-daily injections, respectively (P > 0.2). Furthermore, the mean number of ova/embryos and transferable embryos were higher in cows treated with 400 mg (13.9 ± 1.1 and 6.5 ± 0.7) and 300 mg (12.0 ± 1.0 and 6.1 ± 0.7) of Folltropin®-V than those treated with 200 mg (6.6 ± 0.7 and 4.0 ± 0.5). In summary, superstimulation of Angus donor cows with a single i.m. injection of Folltropin®-V diluted in an SRF resulted in comparable embryo production to the traditional twice-daily i.m. administration of Folltropin®-V over 4 days. Although response did not differ between 300 and 400 mg, results suggest that 300 mg of Folltropin®-V given i.m. by a single or twice-daily injections is the most appropriate dose for Angus donor cows.
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Wock, J. M., L. M. Lyle, and M. E. Hockett. "297 EFFECT OF GONADOTROPIN-RELEASING HORMONE COMPARED WITH ESTRADIOL-17β AT THE BEGINNING OF A SUPERSTIMULATION PROTOCOL ON SUPEROVULATORY RESPONSE AND EMBRYO QUALITY." Reproduction, Fertility and Development 20, no. 1 (2008): 228. http://dx.doi.org/10.1071/rdv20n1ab297.
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Success of a superstimulation protocol is largely dependent on avoiding the effects of the dominant follicle. The objective of this study was to determine if gonadotropin-releasing hormone (GnRH) would serve as a suitable alternative to the traditional estradiol-17β in a superstimulation protocol when using a CIDR. Dairy cattle (n = 411) were superstimulated using one of two treatment protocols. The GnRH-treated animals received a CIDR on random days of the estrous cycle (day 0), and GnRH was injected i.m. (100 micrograms of Cystorelin) on day 3. Porcine FSH (Folltropin-V) was administered twice daily in decreasing doses on days 5 to 8, and on day 8, CIDR were removed and 25 mg of dinoprost (PGF2α) was administered twice. The Estradiol-treated animals received a CIDR and 4 mg estradiol-17β i.m. on random days of the estrous cycle (day 0). Porcine FSH was administered twice daily in decreasing doses on days 4 to 7, and on day 7, CIDR were removed and 25 mg of PGF2α was administered twice. All animals were artificially inseminated at standing estrus and at 6-h intervals during estrus. Animals were inseminated a minimum of twice, and those with extended estrous periods were inseminated three times. Animals not showing estrus were inseminated once 48 h after CIDR removal. Seven days following estrus, embryos were collected. For data analysis, animals were divided into lactating cows, dry cows, and heifers. Dosage of FSH for each animal varied according to previous response rates; therefore, animals were further subclassified as high FSH (≥300 mg in 15 mL) or low FSH (<300 mg in 15 mL). As there were no heifers on high FSH, data analysis was done as a 5 � 2 factorial ANOVA. Total number of ova/embryos, quality, development, and fertilization rates were recorded. Analysis of data showed no statistical differences (P > 0.05) between Estradiol- and GnRH-treated groups for number ova/embryos recovered (9.8 � 0.58 v. 9.7 � 0.65), IETS-grade 1 and 2 embryos (4.7 � 0.37 v. 4.5 � 0.41), percentage fertilized (54.1 � 2.5 v. 53.2 � 2.9), and percentage grade 1 and 2 embryos (49.2 � 2.5 v. 49.3 � 2.8), respectively. No significant interactions were found between treatment and lactation status or treatment and FSH dose. Data suggest that GnRH in combination with a CiDR is an effective means of synchronizing follicle wave emergence in a superstimulation protocol. George Seidel, Josh Walker, John Hassler, and Sam Galphin.
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Garcia Guerra, A., G. A. Bó, J. Villarreal, and G. M. Brogliatti. "295 SUPEROVULATORY RESPONSE IN COWS FOLLOWING SYNCHRONIZATION OF FOLLICLE WAVE EMERGENCE WITH ESTRADIOL AT DIFFERENT STAGES OF THE ESTROUS CYCLE." Reproduction, Fertility and Development 20, no. 1 (2008): 227. http://dx.doi.org/10.1071/rdv20n1ab295.
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Ovarian asynchrony and variability in response to superstimulation remain the most limiting factors in any embryo transfer program (Armstrong D 1993 Theriogenology 39, 7–24). Ovarian response can be increased and less variable if superstimulatory treatment is started at the time of follicular wave emergence (Bö GA et al. 1995 Theriogenology 43, 31–40). A combination of progesterone (P4) and estradiol have been used to synchronize follicular wave for superstimulation. A retrospective analysis was done to compare the ovarian response, superovulatory response and embryo production of cows in Argentina that received progesterone and estradol prior to superstimulation at different stages of the estrous cycle. This research was carried out using different breed of donors (n = 584, 88% Angus) during the last 4 years in Buenos Aires province, Argentina. Heat detection was performed twice a day. At random stages of the estrous cycle, donors received an intravaginal progesterone device (DIB; Syntex, Buenos Aires, Argentina), 2 mg of estradiol benzoate and 50 mg of progesterone (Syntex, Buenos Aires, Argentina) IM on the same day. On day 4 after DIB insertion, superestimulatory treatment was initiated on a decreasing dose regimen of FSH (Pluset; Callier, Spain, or Folltropin, Bioniche Animal Health Inc., Belleville, Ontario, Canada) as IM injections every 12 h over 4 d. On day 6, DIBs were removed, and cows received two doses of 2 mL of cloprostenol 12 h apart. At heat detection, all donors received a dose of 2 mL of GnRH (Dalmarelin; Fatro Von Franken, Buenos Aires, Argentina) by IM injection and were inseminated 12 and 24 h later. Seven days later, embryo collection was performed and ovarian response was evaluated as number of CL + unovulated follicles by transrectal ultrasound using a 7.5-MHz transducer (Pie Medical, Maastricht, the Netherlands). Ova/embryos were evaluated and classified according to the IETS manual. Donors were assigned to receive DIB and estradiol during the following stages of the cycle: group 1: between days 4 and 7 post-estrus (dominant follicle period), group 2: between days 8 and 12 post-estrus (emergence of the second follicular wave), and group 3: between days 13 and 21 post-estrus (dominant follicle of the second wave). Kruskal-Wallis test was used to compare variables among groups, and results are shown in Table 1. Ovarian response as CL + unovulated follicles and number of ovulations were significantly different among groups (P < 0.05). However, there was no significant difference in the number of fertilized ova or transferable embryos. Nevertheless, numeric differences that show that group 2 (started between days 8 and 12 post-estrus) was always superior for all variables. In conclusion, data suggest that estradiol may be more effective in synchronizing follicle wave emergence for superstimulation during the mid-part of the estrous cycle. Table 1. Superovulatory response in cows in which follicle wave emergence was synchronized with estradiol at different stages of the estrous cycle (mean ± SD) Research supported by Centro Genetico Bovino Eolia S.A.
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Dadarwal, D., M. Honparkhe, F. C. F. Dias, T. Alce, C. Lessard, and J. Singh. "Effect of superstimulation protocols on nuclear maturation and distribution of lipid droplets in bovine oocytes." Reproduction, Fertility and Development 27, no. 8 (2015): 1137. http://dx.doi.org/10.1071/rd13265.
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Our objective was to study the effect of superstimulation protocols on nuclear maturation of the oocyte and the distribution of lipid droplets in the ooplasm. Heifers (n = 4 each group) during the luteal phase were either treated with FSH for 4 days (Short FSH), FSH for 4 days followed by 84 h of gonadotropin free period (FSH Starvation) or for 7 days (Long FSH) starting from the day of wave emergence. In all groups, LH was given 24 h after induced luteolysis (penultimate day of FSH) and cumulus–oocyte complexes were collected 24 h later. Oocytes were stained for nuclear maturation (Lamin/chromatin) and lipid droplets (Nile red). The Long FSH group had a greater proportion of mature oocytes (metaphase II) compared with heifers in the Short FSH and FSH Starvation groups (59/100 vs 5/23 and 2/25, respectively; P < 0.01). On average across all groups, oocytes contained 22 pL of lipids (3.3% of ooplasm volume) distributed as 3000 droplets. Average volume of individual lipid droplets was higher in the FSH Starvation (11.5 ± 1.5 10–3 pL, P = 0.03) compared with the Short and Long FSH groups (7.2 ± 0.6 10–3 and 8.0 ± 0.8 10–3 pL, respectively). In conclusion, both FSH Starvation and Short FSH treatments yielded a lower proportion of mature oocytes compared with the Long FSH treatment. Furthermore, FSH starvation led to an accumulation of larger lipid droplets in the ooplasm, indicating atresia. Our results indicate that a longer superstimulation period in beef cattle yields higher numbers and better-quality oocytes.
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Abu-Heija, Adel, and Robert Yates. "Comparison of Controlled Ovarian Superstimulation with or Without Intrauterine Insemination for the Treatment of Unexplained Infertility." Annals of Saudi Medicine 15, no. 5 (September 1995): 464–65. http://dx.doi.org/10.5144/0256-4947.1995.464.
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Mapletoft, Reuben J., and Gabriel A. Bó. "The evolution of improved and simplified superovulation protocols in cattle." Reproduction, Fertility and Development 24, no. 1 (2012): 278. http://dx.doi.org/10.1071/rd11919.
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Superovulation protocols have improved greatly since the early days of bovine embryo transfer when purified gonadotrophins were not available, follicular wave dynamics were unknown physiological phenomena and prostaglandins were not available. Although superstimulatory protocols in cattle are normally initiated mid-cycle, elective control of follicular wave emergence and ovulation have had a great impact on the application of on-farm embryo transfer. However, the most common treatment for the synchronisation of follicular wave emergence involves the use of oestradiol, which cannot be used in many parts of the world. Therefore, the need for alternative treatments has driven recent research. An approach that has shown promise is to initiate follicle-stimulating hormone (FSH) treatments at the time of the emergence of the new follicular wave following ovulation induced by gonadotrophin-releasing hormone. Alternatively, it has been shown that it may be possible to ignore follicular wave status and, by extending the treatment protocol, induce subordinate follicles to superovulate. Finally, the short half-life of pituitary FSH necessitates twice-daily treatments, which are time-consuming, stressful and subject to error. Recent treatment protocols have permitted superstimulation with a single FSH treatment or two treatments 48 h apart, reducing the need for animal handling during gonadotrophin treatments.
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Imai, K., Y. Inaba, H. Yoshioka, Y. Aikawa, M. Ohtake, M. Suzuki, and S. Kobayashi. "206 EFFECT OF FOLLICULAR WAVE SYNCHRONIZATION AND SUPERSTIMULATION ON IN VITRO EMBRYO PRODUCTION." Reproduction, Fertility and Development 20, no. 1 (2008): 182. http://dx.doi.org/10.1071/rdv20n1ab206.
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We previously reported that follicular wave synchronization, by removal of the dominant follicle on Day 5 after ovum pickup (OPU), was effective in increasing oocyte quality in the developing follicles (Imai et al. 2006 32th Annual Conference of the IETS, poster presentation no. 277). The current study was designed to examine the effect of superstimulatory treatment to induce subsequent follicular wave synchronization on embryo production by OPU and IVM-IVF-IVC in Holstein dry cows. Cows were reared under the same feeding and environmental conditions, and 2 OPU sessions were conducted in each cow. In the first session, OPU was performed in 8 cows on arbitrary days of the estrous cycle by using a 7.5-MHz linear transducer with needle (Cova needle, Misawa Medical, Tokyo, Japan) connected to an ultrasound scanner (SSD-1200, Aloka, Tokyo, Japan). Follicles larger than 8 mm in diameter were then aspirated and a CIDR was inserted on Day 5 (the day of first OPU session = Day 0). Cows then received 30 mg of FSH (Antrin-R10; Kawasaki Mitaka Pharmaceutical Co., Tokyo, Japan) twice a day from Days 7 to 10 in decreasing doses (6, 6, 4, 4, 3, 3, 2, 2 mg) by i.m. injection. Cloprostenol (PGF; Clopromate C; Sumitomo Pharmaceuticals Co., Tokyo, Japan; 0.75 mg) was administered in the morning of Day 9 (third day of superstimulation). The second OPU session was performed 48 h after PGF administration (Day 11), and only follicles larger than 5 mm in diameter were aspirated. The CIDR was removed from the cows just before OPU. Collected oocytes were evaluated by their cumulus cell morphology, cytoplasmic color, and density. Grades 1 and 2 COC were matured, fertilized, and cultured as described by Imai et al. [2006 J. Reprod. Dev. 52(Suppl.), S19–S29]. Embryo development was assessed by the cleavage rate on Day 2 and by the blastocyst formation rate on Days 7 to 8 (the day of insemination = Day 0). Data were analyzed by Student's t-test. There were no differences in the mean (� SD) number of aspirated follicles or collected oocytes between the first (32.5 � 6.8 and 26.0 � 12.7, respectively) and second (29.3 � 10.4 and 19.0 � 9.4, respectively) OPU sessions (P > 0.1). The percentage of Grade 1 and 2 oocytes for the second OPU session (90.5 � 13.8%) was significantly higher (P < 0.01) than for the first OPU session (63.1 � 6.3%), and significant differences were found for cleavage (79.4 � 14.1, 61.8 � 25.1, P < 0.01) and blastocyst rates (68.1 � 16.7, 24.2 � 22.3, P < 0.001) between sessions. The mean numbers of blastocysts obtained per session were 4.3 � 2.9 and 12.8 � 8.7 in the first and second sessions, respectively (P < 0.01). These results indicate that superstimulatory treatment and subsequent follicular wave synchronization were effective on in vitro embryo production by increasing the oocyte quality.
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Lv, X. Q., J. H. Xue, Y. L. Zhu, H. B. Liang, and B. H. Xuan. "207 EFFECT OF REPEAT HORMONE STIMULATION ON THE YIELD AND IN VITRO DEVELOPMENT OF JUVENILE CALF OOCYTES." Reproduction, Fertility and Development 26, no. 1 (2014): 217. http://dx.doi.org/10.1071/rdv26n1ab207.
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Juvenile in vitro embryo transfer can markedly reduce animal generation intervals. The purpose of this study was to investigate the ovarian response of juvenile calves and in vitro oocyte developmental capacity after superstimulation. Experiments on calves were performed in accordance with the Animal Welfare Regulations. A total of 36 donor juvenile calves on standard nutrition and in a disease-free environment, were selected from the breeding farm of the Beijing Dairy Cattle Center. At 60 days of age, calves were randomly assigned into three groups of four calves each, replicated three times. On day 1, Group 1 received a progesterone vaginal insert (CIDR, 300 mg per device); Group 2 received a CIDR and 0.5 mg oestrogen benzoate (China); Group 3 received a CIDR, 0.5 mg oestrogen benzoate, and 50 mg progesterone (China). Then, calves were injected with FSH (Folltropin-V, Bioniche Animal Health, Belleville, ON, Canada) twice daily on days 5 (40 mg/40 mg) and 6 (30 mg/30 mg) at 12 h intervals. Cumulus–oocyte complexes (COCs) were recovered from the superstimulated calves 12 to 14 h after the final FSH treatment. COCs were considered usable unless they were damaged or had expanded cumulus layers. Usable COCs were matured in vitro for 24 h in maturation medium consisting of TCM199, 10% FBS, 10 μg mL–1 FSH, 1 μg mL–1 LH, 1 μg mL–1 E2–17β, 100 IU mL–1 penicillin, 100 μg mL–1 streptomycin, with (+Cys) or without (–Cys) 100 μM Cysteamine. Each calf oocyte was cultured in one well. The final concentration added to each fertilization drop was 5 × 106 sperm mL–1. Sperm and oocytes were co-cultured in IVF-100 medium (BO liquid+10 μg mL–1 heparin, Japan) at 38.5°C, 5% CO2 and a saturated humidity for 6 to 8 h. Blastocyst production rates were determined after 7 and 8 d of in vitro culture in CR1aa medium without the addition of cysteamine. Differences among treatments in each experiment were determined by one-way ANOVA and a multiple range test. Superstimulatory results indicated that more follicles were aspirated (63.2 per calf) and more usable oocytes were recovered (48.0 per calf) in Group 1 than in the other two groups (Group 2–45.2 and 31.8, respectively; Group 3–35.4 and 28.3, respectively; P < 0.05). No difference was observed between Groups 2 and 3. Superstimulation of calves twice at 30 day intervals in Group 2 (n = 12) did not affect the number of follicles or usable oocytes (overall, 44.2 and 28.0 per calf). Maturation rates (86.5% v. 85.0%, respectively) and cleavage rates (84.4% v. 80.0%, respectively) did not differ whether cysteamine was not (–Cys; n = 318) or was (+Cys; n = 330) added to the maturation medium. However, the blastocyst rate differed significantly (12.9% v. 35.2%, respectively; P < 0.01). This study established a protocol for the superstimulation of juvenile calves with an average of 48 oocytes obtained per calf. Superstimulation and surgical oocyte recovery twice at an interval of 30 days had no adverse effect on follicle development or oocyte recovery. The novelty of this research is that the blastocyst production rate of calf oocytes (35.2%) in maturation medium supplemented with cysteamine was similar to that reported in the cow.
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Son, Dong-Soo, Chang-Yong Choe, Sun-Ho Choi, Sang Rae-Cho, Hyun-Jong Kim, Man-Hye Han, Il-Sun Ryu, Guk-Hyun Suh, Ui-Hyung Kim, and Ill-Hwa Kim. "Effect of estradiol benzoate or GnRH treatment prior to superstimulation in CIDR-treated, Korean native cows (Bos taurus)." Animal Reproduction Science 100, no. 1-2 (July 2007): 14–21. http://dx.doi.org/10.1016/j.anireprosci.2006.06.005.
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Son, D. S., C. Y. Choe, S. R. Cho, S. H. Choi, H. J. Kim, and I. H. Kim. "390 THE EFFECT OF REDUCED FSH DOSE AND NUMBER OF TREATMENTS ON SUPEROVULATORY RESPONSE IN CIDR-TREATED KOREAN NATIVE COWS." Reproduction, Fertility and Development 19, no. 1 (2007): 310. http://dx.doi.org/10.1071/rdv19n1ab390.
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Reducing the total dose and numbers of treatments with FSH for superstimulation without decreasing embryo yield may be less stressful and more economical for bovine embryo transfer. The objective of this study was to investigate the effect of dose and the number of days of FSH treatment on superovulatory responses in CIDR-treated Korean native cows. Forty-two cows, at random stages of the estrous cycle, received a CIDR device (CIDRTM; InterAg, Hamilton, New Zealand), 1 mg estradiol benzoate (SY Esrone; Samyang, Seoul, Korea) and 50 mg progesterone (SY Ovaron; Samyang); gonadotropin treatment began 4 days later. Cows were divided into 2 groups based on the dose and numbers of days of treatment with porcine FSH (pFSH): T1 group (n = 20): a total of 28 mg pFSH (recommended dose of Antorin�; Kawasaki Pharmaceutical, Tokyo, Japan) was given in twice daily IM injections in decreasing doses over 4 days (5, 5, 4, 4, 3, 3, 2, and 2 mg); and T2 group (n = 22): a total of 24 mg pFSH given in twice daily decreasing doses over 3 days (5, 5, 4, 4, 3, and 3 mg). Otherwise, all cows received the same treatments. Twenty-five and 15 mg dinoprost (PGF2α; Lutalyse; Pharmacia & Upjohn, Puurs, Belgium) were given with the 5th and 6th injections of pFSH, respectively. CIDR devices were withdrawn with the 6th pFSH injection, and the cows received 100 �g Gonadorelin (GnRH; Fertagyl; Intervet, Boxmeer, The Netherlands) 36 h after CIDR device removal. Cows were artificially inseminated using commercial semen from 4 Korean native bulls twice, at 48 and 60 h after CIDR device removal, and embryos were recovered 6 or 7 days after the 2nd insemination. The number of CL was counted on the day of embryo recovery by transrectal ultrasonography (Sonovet 600 with 5.0 MHz linear-array transducer; Medison Co., Ltd., Seoul, Korea). The recovered embryos were evaluated according to the IETS Manual for stage of development and quality. All data between groups were compared using Student's t-test. The numbers of CL (9.7 � 1.1 vs. 9.4 � 1.3), total ova/embryos (7.2 � 1.1 vs. 6.3 � 1.4), transferable embryos (4.4 � 1.0 vs. 3.6 � 0.9), degenerate embryos (0.9 � 0.3 vs. 1.3 � 0.4), and unfertilized ova (2.0 � 0.6 vs. 1.5 � 0.5) did not differ between groups (T1 vs. T2), respectively (P > 0.05). Data indicate that the reduced dose (24 vs. 28 mg) and numbers of treatments (6 vs. 8) of pFSH for superstimulation of Korean native cows does not affect the embryo yield.
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Rogan, D., A. Tríbulo, H. Tríbulo, R. Tríbulo, D. Carballo Guerrero, P. Tríbulo, R. J. Mapletoft, and G. A. Bó. "416 DOSE TITRATION FOR SUPERSTIMULATION OF BRANGUS AND BONSMARA DONORS WITH Folltropin®-V BY A SINGLE INTRAMUSCULAR INJECTION." Reproduction, Fertility and Development 22, no. 1 (2010): 365. http://dx.doi.org/10.1071/rdv22n1ab416.
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Two experiments were designed to evaluate the superovulatory response of Brangus and Bonsmara donor cows to different dosages of Folltropin®-V (Bioniche Animal Health Inc., Belleville, Ontario, Canada) given by a single i.m. injection or twice-daily i.m. injections. In Experiment 1, Brangus cows (n = 12) were superstimulated by 6 treatments (2 × 3 factorial) in a crossover design (i.e. all cows received the 6 treatments and all treatments were represented on each day). On Day 0, cows received 5 mg of estradiol-17β plus 50 mg of progesterone and a Cue-Mate® (Bioniche Animal Health Inc.). On Day 4, cows were superstimulated with 300, 260, or 200 mg of NIH-FSH-P1 Folltropin®-V (Bioniche Animal Health Inc.) in twice-daily decreasing doses over 4 days or diluted in a slow release formulation (SRF; Bioniche Animal Health) and given in a single i.m. injection. The single injection was prepared by diluting the Folltropin®-V lyophilized powder in 1 mL of saline followed by mixing with 9 mL of the SRF in the syringe immediately before administration. In the am and pm of Day 6, all cows received PGF2, and Cue-Mates® were removed in the pm. Cows received 12.5 mg of porcine LH (Lutropin®-V; Bioniche Animal Health Inc.) in the am of Day 8 and were inseminated 12 and 24 h later. Ova/embryos were collected on Day 15 and data were analyzed by ANOVA. There was no effect of treatment (i.e. single v. twice-daily injections; P > 0.2) nor a treatment by dosage interaction (P < 0.6) on the mean (± SEM) number of total ova/embryos or transferable embryos (13.1 ± 1.9 and 7.5 ± 1.2 v. 15.5 ± 1.7 and 7.6 ± 1.0 for single v. twice-daily injections, respectively). The total number of ova/embryos did not differ among Folltropin®-V dosages (15.0 ± 2.3, 15.7 ± 2.0, and 12.1 ± 2.5 for 300, 260, and 200 mg, respectively; P > 0.4). However, the number of transferable embryos tended (P < 0.09) to be higher in donors receiving 260 mg (9.5 ± 1.6) than 200 mg (5.2 ± 0.8), with 300 mg (7.9 ± 1.5) intermediate. In Experiment 2, Bonsmara cows (n = 16) were superstimulated by 4 treatments (2 × 2 factorial) in a crossover design similarly to Experiment 1, except that 2 dosages of Folltropin®-V (200 and 300 mg) were evaluated. There were no significant effects of dosage of Folltropin®-V (P > 0.9), treatment (P > 0.3), or interaction (P < 0.4) on embryo production. The total number of ova/embryos and transferable embryos were 11.9 ± 2.0 and 7.2 ± 1.1 v. 11.1 ± 1.1 and 7.6 ± 0.7 for single and twice-daily injections, respectively, and 11.9 ± 1.9 and 7.6 ± 1.0 v. 11.1 ± 1.3 and 7.2 ± 0.8 for 300 and 200 mg of Folltropin®-V, respectively. Superstimulation of Brangus and Bonsmara cows with a single i.m. injection of Folltropin®-V diluted in a SRF resulted in comparable embryo production to twice-daily administration of Folltropin®-V over 4 days. While 260 mg seems to be the most appropriate dosage for Brangus donors, 200 mg seems to be adequate for Bonsmara donors.
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Foster, B. A., E. J. Gutierrez, and K. R. Bondioli. "15 The effect of different bovine oocyte recovery methods on oocyte ultrastructure pre- and post-in vitro maturation." Reproduction, Fertility and Development 31, no. 1 (2019): 133. http://dx.doi.org/10.1071/rdv31n1ab15.
Full textAbstract:
In current in vitro embryo production practices, method of oocyte recovery tends to be based primarily on the personal preference of the operators and availability of resources rather than on any practical understanding of the best practices for retrieving oocytes of the greatest competence for embryo production. There is little consensus on the effects of either postmortem follicular atresia or exogenous hormone-stimulated follicle development on the quality of oocytes, but it is expected that both interventions have effects. Cumulus-oocyte complexes were collected from beef cattle using 3 methods: unstimulated ovum pickup (OPU), OPU following superstimulation (FSH), and oocyte aspiration from postmortem-recovered ovaries (PM). Each collection method was repeated 3 times, with 25 individuals used in each. There were no interventions in place before oocyte recovery in either OPU or PM. In FSH, cows underwent dominant follicle removal and were treated with 200mg of FSH in 6 doses before OPU. The majority of oocytes (66 OPU, 80 FSH, and 160 PM) were fertilized and cultured, with a portion being reserved for transmission electron microscopy (2 per treatment at each of germinal vesicle and metaphase II) and a portion being reserved for a complementary study. However, oocytes collected by OPU following FSH had notable ultrastructure differences from those collected by other methods, suggesting a potential effect of follicle stimulation on both the development and cytoplasmic maturation of oocytes. Although oocytes from all recovery methods and at both maturation levels exhibited mitochondria primarily in the hooded conformation, both OPU and FSH oocytes, at germinal vesicle and metaphase II, had several anomalous mitochondria exhibiting a lobulated, budding-type shape. This is potentially indicative of these oocytes having failed to complete the mitochondrial division phase before recovery. An average of 11.6, 10.6, and 11.9 oocytes per individual were recovered from PM, FSH, and OPU, respectively, resulting in oocyte recovery of 55, 61, and 57% of aspirated follicles, respectively (P>0.05). Fewer oocytes from OPU developed to morulas compared with other recovery methods (chi-square; P<0.01; 24% PM, 21% FSH, and 3% OPU). Cleaved embryos from OPU had decreased blastocyst production rates at 14% compared with 59% (PM) and 55% (FSH; chi-square; P<0.01). Overall results suggest that although superstimulation may affect oocyte ultrastructure, this does not have a negative effect on oocyte competence, though further investigations are required. Oocyte recovery rates were not affected by recovery method, but oocytes recovered by unstimulated OPU had decreased developmental competence compared with those recovered either postmortem or following superstimulation.
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Palomino, J. M., R. B. McCorkell, M. R. Woodbury, M. P. Cervantes, B. M. Toosi, and G. P. Adams. "321 OVARIAN SUPERSTIMULATION AND OOCYTE COLLECTION IN WOOD BISON, A THREATENED CANADIAN SPECIES." Reproduction, Fertility and Development 23, no. 1 (2011): 257. http://dx.doi.org/10.1071/rdv23n1ab321.
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The wood bison (Bison bison athabascae) is a threatened species, and recovery is constrained by endemic tuberculosis and brucellosis in wild herds in and around Wood Buffalo National Park, Alberta, Canada. The Committee on the Status of Endangered Wildlife in Canada has recommended the eradication of affected herds and repopulation with healthy bison obtained through the use of reproductive technologies. As part of the conservation effort, the specific objective of this study was to develop an effective ovarian superstimulatory protocol in bison that will permit collection of a large number of oocytes for the ultimate purpose of in vitro embryo production. Ovarian function was synchronized among female wood bison during the ovulatory season (December) by giving a luteolytic dose of prostaglandin followed 8 days later by transvaginal ultrasound-guided follicular ablation. On the day after follicular ablation (expected day of follicular wave emergence, Day 0), bison were assigned randomly to 2 groups (n = 11/group) and given either a single intramuscular dose of 2500 IU of eCG or subcutaneous doses of 200 mg of FSH given on Days 0 and 2. A luteolytic dose of prostaglandin was given on Day 2, and 25 mg of LH was given intramuscularly on Day 4. Cumulus–oocyte complexes (COC) were collected 24 h after LH treatment by transvaginal ultrasound-guided follicle aspiration using an 18 G × 2’ aspiration needle attached to a flexible tubing and a regulated vacuum pump (flow rate, 22 mL min–1). The COC were classified morphologically from 1 (excellent) to 4 (poor) based on attributes of the ooplasm and the number of surrounding cumulus cells. The number of COC in grade 1 to 3 was combined (i.e. grades acceptable for IVF) for purposes of statistical interpretation. Ovarian response and oocyte quality were compared between groups by t-tests, and COC collection rate was compared by chi-square analysis. The number (mean ± SEM) of follicles ≥5 mm was greater (P < 0.05) in bison treated with FSH than in those treated with eCG (12.2 ± 1.7 v. 5.8 ± 0.5), resulting in a greater number of follicles aspirated (11.2 ± 1.8 v. 5.6 ± 0.5; P < 0.05), more COC collected (7.2 ± 1.4 v. 3.4 ± 0.6; P < 0.05), and more COC of acceptable quality (6.1 ± 1.4 v. 2.5 ± 0.5). The COC collection rate (i.e. number of COC collected per number of follicles aspirated), however, did not differ between FSH- and eCG-treated groups [79/123 (64%) v. 37/62 (60%)]. The results document the successful collection of viable oocytes from superstimulated wood bison via tranvaginal ultrasound-guided follicle aspiration. The number of large follicles, follicles aspirated, and COC collected were more than twice as high in bison treated with FSH than in those treated with eCG. This work was supported by grants from Advancing Canadian Agriculture and Agri-Food Fund and Agri-Food Innovation Fund. The authors thank Bioniche Animal Health Canada for providing Folltropin, Lutropin, and Pregnecol.
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Patino, Cristian, Eduardo Arroyo, Michela Ciccarelli, Jacobo Rodriguez, Alan Conley, and Ahmed Tibary. "Serum anti-Müllerian hormone concentrations in female alpacas: variations during the reproductive cycle and correlation with ovarian superstimulation response." Clinical Theriogenology 14, no. 2 (June 1, 2022): 91–97. http://dx.doi.org/10.58292/ct.v14.9139.
Full textAbstract:
Earlier, we validated an anti-Müllerian hormone (AMH) enzyme-linked immunosorbent assay kit for male alpacas. First, wecompared the validation data with another kit. There was a high correlation (R2 = 0.94) between these 2 kits. Second, we used thelatter kit to determine serum AMH concentrations during follicular and luteal phases of the reproductive cycle in female alpacas.There were no differences (p = 0.39) in serum AMH concentrations in alpacas (n = 11) between peak follicular and luteal phases(mean ± SEM, 1.33 ± 0.35 versus 1.18 ± 0.34 ng/ml, respectively). Third, we treated female alpacas (n = 13; 5 - 11 years) after 14-day treatment with decreasing doses of porcine follicle-stimulating hormone. There was no effect (p > 0.05) of day of treatmenton serum AMH concentrations. Number of follicles (7 - 10 mm; mean ± SD [as determined via transrectal ultrasonography]) atend of treatment (12.69 ± 5.25; range: 6 - 24) was positively correlated (R2 = 0.7; p < 0.01) with serum AMH concentrations. Toconclude, the kit tested is usable for female alpacas; serum AMH concentrations were not affected by the cycle stage nor by ovariansuperstimulation treatment. Furthermore, a significant correlation between serum AMH serum concentrations and response to superstimulationsuggested that estimation of serum AMH concentrations may be valuable in determining ovarian follicular reserve.
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Zwiefelhofer, E. M., A. R. T. Krause, L. B. Araujo, R. J. Mapletoft, and G. P. Adams. "198 Superovulatory response in cows undergoing aromatase inhibitor treatment." Reproduction, Fertility and Development 31, no. 1 (2019): 224. http://dx.doi.org/10.1071/rdv31n1ab198.
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The aromatase inhibitor letrozole has been used in the treatment of infertility in women by inducing mild ovarian superstimulation or augmenting the ovarian response to FSH treatment. The effect has been attributed to an apparent up-regulation of FSH receptors on granulosa cells as a result of increased androgens (Weil et al. 1999 J. Clin. Endocrinol. Metab. 84, 2951-2956). The objective of this study was to determine whether letrozole will augment the superstimulatory response in FSH-treated cattle. Mature, non-lactating Holstein cows (n=30) were given 2 luteolytic doses of PGF2α 12h apart and scanned daily by transrectal ultrasonography to detect ovulation. Transvaginal ultrasound-guided ablation of follicles ≥5mm was performed in all cows as a group, at 5 to 8 days after ovulation. On the day of ablation, cows were assigned randomly to 2 groups (n=15 per group) and given either an intravaginal letrozole-releasing device or a blank device for 5 days. Coinciding with expected wave emergence (Day 0), cows in both groups were given 8 doses of 50mg of pFSH (Folltropin; Vetoquinol, Lure, France) IM at 12-h intervals, and PGF2α on Days 3 and 3.5. At the time of the second PGF2α, vaginal devices were removed and paint was applied to the tailhead to facilitate detection of oestrus. On Day 5, cows were given gonadotropin-releasing hormone (100mg gonadorelin) IM and artificially inseminated 12 and 24h later. Ova/embryos were collected by transcervical uterine flush on Day 12. The ovaries were examined by ultrasonography on Day 0, 3.5, 5, 6.5, and 12 to record the follicular and luteal response. Nominal data were compared between groups by t-test and by ANOVA for repeated-measures and are expressed as mean±s.e.m. Binomial data were compared by chi-squared test. The number of follicles at wave emergence did not differ between letrozole and control groups (24.6±3.1 and 26.5±3.3 respectively; P=0.68). There was no treatment effect or treatment×day interaction in the number of follicles >8mm on Day 3.5, 5, and 6.5. Letrozole-treated cows exhibited oestrus later than controls (50.3±1.1h v. 40.7±2.0h after first PGF2α; P<0.001) and had less variance in interval to oestrus (residuals, 3.1±0.48h v. 6.7±0.87 h; P<0.01). The number of CL on Day 6.5 was lower in the letrozole group than in the control group (9.1±1.1v. 12.3±1.1; P=0.05). The proportion of ovulations (number of CL on Day 12 over number of follicles ≥3mm on Day 0) was lower in the letrozole group than in the control group (0.65±0.05v. 0.82±0.08; P=0.02). The total number of ova/embryos collected per cow did not differ between letrozole and control groups (5.0±1.9v. 5.4±1.8; P=0.75), nor did the number of transferable embryos differ (3.0±1.2v. 4.3±1.5; P=0.56). In conclusion, treatment with a letrozole-releasing device during ovarian superstimulation did not improve the superovulatory response or embryo collection rate in mature Holstein cows, but letrozole treatment resulted in more synchronous oestrus, which may be used in the design of a fixed-time AI protocol following superstimulatory treatment in cattle. This research was supported by the Alberta Livestock and Meat Agency.
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Imai, K., T. Somfai, M. Ohtake, Y. Inaba, Y. Aikawa, S. Sugimura, H. Yoshioka, S. Kobayashi, and K. Konishi. "278 EFFECT OF FOLLICULAR WAVE SYNCHRONIZATION AND SUPERSTIMULATION ON THE NORMALITY OF BOVINE EMBRYOS PRODUCED IN VITRO." Reproduction, Fertility and Development 22, no. 1 (2010): 296. http://dx.doi.org/10.1071/rdv22n1ab278.
Full textAbstract:
We previously reported that follicular wave synchronization by dominant follicle removal on Day 5 and the start of a superstimulatory treatment on Day 7 after ovum pick-up (OPU) was effective to increase oocyte quality (Imai et al. 2008 Reprod. Fertil. Dev. 20, 182). The present study was designed to examine the effect of superstimulatory treatment-induced follicular wave synchronization on quality of embryos obtained by OPU and in vitro production. Japanese Black cows were reared under the same feeding and environmental conditions and 2 OPU sessions were conducted in each cow. The first OPU session was performed in 7 cows at arbitrary days of estrous cycle using a 7.5-MHz linear transducer with needle connected to an ultrasound scanner. Then, follicles larger than 8 mm in diameter were aspirated and CIDR was inserted on Day 5 (the day of first OPU session = Day 0). The cows then received 30 mg of FSH twice a day from Days 7 to 10 in decreasing doses (4, 4, 3, 3, 2, 2, 1, 1 mg per shot) by i.m. injections. Cloprostenol (PGF; 0.75 mg) was administered in the morning of Day 9. The second OPU session was performed 48 h after PGF administration (Day 11) and only follicles larger than 5 mm in diameter were aspirated. The CIDR was removed from the cows just before OPU. Grade 1 and 2 cumulus oocyte complexes were in vitro matured, fertilized (IVF), and cultured as described by Imai et al. (2006 J. Reprod. Dev. 52, Suppl. S19-29). Some zygotes were fixed and stained to check their sperm penetration. Embryo development was monitored by time-lapse cinematography for 168 h after IVF. Cleavage pattern of embryos was classified morphologically into normal and abnormal (including those with multiple fragments, protrusions, 3 to 4 blastomeres, and uneven cell division) groups at their first cleavage. Normal penetration rate of second OPU session was significantly (P < 0.05) higher than that of the first OPU session. There were no differences in the mean percentage of total blastocyst and grade 1 blastocyst rates between the first (45.2 and 46.9%, respectively) and second (47.5 and 41.8%, respectively) OPU sessions. However, the rates of blastocysts developing from embryos that were beyond the 4-cell stage at 48 h after IVF was significantly (P < 0.05) higher after the second OPU session (81.2%) than after the first OPU session (67.4%). Furthermore, a significant difference (P < 0.05) was found in the rates of normal cleavage at the first cell division in embryos that developed to the blastocyst stage between the first and second OPU sessions (53.3% and 73.9%, respectively). These results indicate that superstimulatory treatment-induced follicular wave synchronization improved the normality of fertilization and development of cattle oocytes obtained by OPU. This work was supported by the Research and Development Program for New Bio-industry Initiatives.
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Brogliatti, G., J. Villarreal, L. Cutaia, A. Albrech, A. Garcia Guerra, and G. A. Bó. "290 OPTIMAL OF DOSE OF OVINE PITUITARY GLAND EXTRACT OVAGEN® FOR SUPERSTIMULATION OF BEEF COW DONORS IN ARGENTINA." Reproduction, Fertility and Development 21, no. 1 (2009): 242. http://dx.doi.org/10.1071/rdv21n1ab290.
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The success of an embryo transfer program is measured by the number of calves born alive by female donor in a given period. The success is influenced by several factors related to the number of ovulations, fertilization rate, and embryo viability (Armstrong D 1993 Theriogenology 39, 7–24). One of the main inconveniences of embryo transfer programs is the variability of superovulatory response to treatments. An experiment was designed to determine the optimal dose of ovine pituitary gland extract Ovagen® (ICPbio, Ltd., Auckland, New Zealand) for inducing superovulation in Aberdeen Angus donor cows. Sixty cycling multiparous donors with a condition score between 3.5 and 4.5 (Scale 1 to 5) were used. All donors received an intravaginal progesterone device DIB (1 g of Progesterone, Syntex, Buenos Aires, Argentina), along with 2 mg of EB (Estradiol Benzoate, Syntex, Buenos Aires, Argentina) and 50 mg of progesterone (Laboratorio Rio de Janeiro, Argentina) on Day 0. Superestimulatory treatment began on Day 4 and donors were randomly assigned to 3 treatment groups according to the total dose of NIADDK-oFSH-Z (Ovagen®) as follows: Group 1, cows received the total dose recommended by the manufacturer (100%* Group), 17.6 mg; Group 2, cows received 75% of the total dose recommended by manufacturer (75% Group), 13.2 mg and Group 3, cows received 50% of the total dose recommended by manufacturer (50% Group), 8.8 mg. All cows received two 150 μg of D+ cloprostenol IM (Ciclase, Syntex) injections on Day 6 given at 12 h interval. DIB was removed on Day 7 a.m. On Day 8 a.m., cows received 0.05 mg of GnRH IM (Gonasyn, Syntex). Fixed time AI was done on Day 8 p.m. and Day 9 a.m. with high quality frozen–thawed semen. On Day 15 embryo collection was performed by non-surgically method and evaluated by developmental stage and quality. The efficiency of superestimulatory response was evaluated by total amount of collected ova-embryos, fertilized ova and embryos Grade 1, 2, and 3, (according to IETS manual). One way AOV test was used to compare variables among groups and results are shown in Table 1. A significant increase in total CL was observed for the groups receiving 75% or 100% of the recommended dose with respect to the group receiving 50% of that dose. There was also a significant greater number of grade 1 embryos for the groups receiving 75% or 100% of the dose than in the group receiving 50% of the recommended dose. There were no differences for any of the evaluated parameters between 75% and 100% dose groups. These results suggest that acceptable superstimulatory responses can be obtained using reduced doses of Ovagen® in Aberdeen Angus donor cows. (*) the percentage makes reference to the total dose recommended by the laboratory of origin. Table 1.Embryo collection results of superstimulated donors with different dose* of NIADDK-oFSH-Z (Ovagen) Research supported by Syntex S.A. and Eolia S.A.
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Sala, R. V., L. C. Carrenho-Sala, M. Fosado, E. Peralta, D. C. Pereira, D. Moreno, J. F. Moreno, and A. Garcia-Guerra. "135 Use and dose of porcine follicle-stimulating hormone for ovarian superstimulation prior to ovum pickup and in vitro embryo production in pregnant Holstein heifers." Reproduction, Fertility and Development 31, no. 1 (2019): 192. http://dx.doi.org/10.1071/rdv31n1ab135.
Full textAbstract:
The benefit of superstimulation with exogenous FSH before ovum pickup for in vitro embryo production has been the subject of significant controversy. In addition, there is limited information on different dose regimens. Thus, the objective of the present study was to evaluate the effect of dose of porcine (p)-FSH during superstimulation before ovum pickup (OPU) on in vitro embryo production in pregnant heifers. Pregnant Holstein heifers (n=36) were assigned to a complete 3×3 crossover design. Three treatment groups were evaluated as follows: p-FSH 0mg (FSH0), p-FSH 160mg (FSH160) and p-FSH 300mg (FSH300). Three sessions of OPU were performed on each animal at 48, 62 and 76 days of gestation, with a washout interval between sessions of 14 days. Follicular wave emergence was synchronized by dominant follicle removal. Heifers in the FSH0 group received no further treatment, whereas the remaining groups received a total of 4 injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3 and 32.0mg) or FSH300 (90.0, 80.0, 70.0 and 60.0mg), beginning 36h after dominant follicle removal. Ovum pickup was performed in all heifers 40h after the last p-FSH injection. Heifers were subjected to OPU for oocyte recovery, and number of follicles was determined. Recovered oocytes were processed and in vitro embryo production performed. Differences between treatment groups were evaluated by generalized linear mixed models. Data are presented (Table 1) as mean±standard error of the mean. There was no effect of days in gestation for any of the outcomes evaluated (P>0.05). Follicle numbers at the time of oocyte recovery were different (P<0.01) between groups. Heifers in the FSH300 group had a greater (P<0.05) number of medium, large and total follicles than heifers in the FSH0 group, whereas heifers in the FSH160 were intermediate. Total number of recovered, viable and cleaved oocytes were greater (P<0.01) in FSH300- than in FSH160- and FSH0-treated heifers. Cleavage rate and blastocyst development rate were not different (P>0.10) between groups. The number of grade 1 and 2 blastocysts was greater in FSH300- than in FSH160- and FSH0-treated heifers (P<0.03). In summary, the use of 300mg of p-FSH before OPU in pregnant heifers increases the number of follicles, oocytes and blastocysts produced per heifer with no detrimental effect on oocyte competence. Table 1.Ovum pickup and in vitro embryo production in pregnant heifers treated with different doses of porcine FSH
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Carrenho-Sala, L., M. Fosado, R. Sala, E. Peralta, D. Pereira, D. Moreno, J. Moreno, and A. Garcia-Guerra. "176 Synchronisation of follicle wave emergence prior to superstimulation with purified FSH for ovum pickup affects blastocyst rate in pregnant Holstein heifers." Reproduction, Fertility and Development 32, no. 2 (2020): 215. http://dx.doi.org/10.1071/rdv32n2ab176.
Full textAbstract:
The timing of initiation of superstimulatory treatments relative to follicle wave emergence has been shown to affect ovulatory response and invivo embryo production. The significant increase of invitro embryo production (IVP) and the possibility of using pregnant animals as oocyte donors has created the need to optimise superstimulatory treatments for IVP in pregnant cattle. Thus, the objective of the present study was to evaluate the effect of synchronisation of follicle wave emergence before superstimulation for ovum pickup (OPU) and IVP in pregnant heifers. Pregnant (47-62 days of gestation) Holstein heifers (n=28) 19.5±0.3 months of age were assigned in a completely randomised design to one of two groups: synchronisation of follicular wave emergence by dominant follicle removal (DFR; all follicles >6mm) or untreated control (no DFR). Superstimulatory treatments were initiated 36h after DFR or at random stages of the follicular wave in the no-DFR group and consisted of the administration of 160mg of purified FSH (Folltropin-V, Vetoquinol) over four injections 12h apart as follows: 48.0, 42.7, 37.3, and 32.0mg. Ovum pickup was performed in all heifers 40h after the last purified FSH injection. Heifers were subjected to OPU for oocyte recovery, and the number of follicles was determined. Recovered oocytes were processed in groups by treatment, and IVP was performed. Differences between treatment groups were evaluated using generalised linear mixed models. Results are presented in Table 1 and are expressed as means±s.e.m. for data collected at the time of OPU or as proportions for embryo production results. The number of small follicles (<6mm) at the time of OPU was greater in the no-DFR group than in the DFR group (P=0.04). Conversely, there were no differences between treatments in the number of medium follicles (6-10 mm; P=0.17), large follicles (>10 mm; P=0.11), total follicles (P=0.93), total number of recovered oocytes (P=0.4), or number of viable oocytes (P=0.53). The mean oocyte percentage recovery rate was not different between heifers in the DFR (53.6±4.7%) and no-DFR (56.5±4.7%) groups (P=0.52). Both cleavage and blastocyst rate were greater (P<0.008) in the DFR group than in the no-DFR group; as a result, the number of transferable embryos per animal was 5.6 in the DFR group and 2.8 in the no-DFR group. In summary, initiation of superstimulatory treatments at the time of follicle wave emergence improves cleavage and blastocyst rates, thus leading to greater embryo production. Table 1.Ovarian response and embryo production in pregnant heifers superstimulated with or without synchronisation of follicle wave emergence Variable DFR No DFR Small follicles, n 8.1±1.2A 12.1±1.8B Medium follicles, n 18.3±1.3 13.7±2.0 Large follicles, n 2.4±0.6 1.4±0.4 Total follicles, n 28.8±1.4 27.2±2.2 Total oocytes, n 15.4±1.5 16.0±1.9 Viable oocytes, n 13.7±1.5 13.4±1.8 Cleavage rate,% (n) 77.1 (192)A 64.4 (188)B Blastocyst rate,% (n) 40.6 (192)A 20.7 (188)B A,BMeans within a row with different superscripts differ (P<0.05).
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Konrad, J., J. A. Berdugo Gutierrez, R. Yuponi, N. Vallejos, W. Cardona-Maya, G. Clérico, G. Crudeli, and M. Sansinena. "224 Superstimulation effect on invitro embryo production and relationship with anti-Müllerian hormone in buffaloes (Bubalus bubalis) aspirated 7 or 14 days after follicular ablation." Reproduction, Fertility and Development 32, no. 2 (2020): 240. http://dx.doi.org/10.1071/rdv32n2ab224.
Full textAbstract:
The success of buffalo invitro embryo production (IVEP) depends, in part, on the number and competence of oocytes. Superstimulation with FSH has been linked to oocyte quality and embryo yield, and anti-Müllerian hormone (AMH) has been proposed as an indicator of ovarian reserve. The objective was to evaluate FSH stimulation in one ovum pick-up (OPU) cycle conducted 7 or 14 days after follicular ablation; we additionally analysed residual FSH treatment effect carried over to a subsequent aspiration cycle. Finally, we evaluated the relationship between plasma AMH and FSH response. Nine multiparous, cycling Murrah donors (603±7.0kg, body condition score=3.5) were subjected to complete follicular ablation followed by OPU 1 week later (T0 Control), and plasma AMH was determined at this point. They were then randomly allocated to FSH stimulation (3 days b.i.d. in descending doses, total 300mg of pFSH, Vetpharm, with the last injection 48h before OPU) with aspiration conducted 7 (T7 FSH, n=5) or 14 days (T14 FSH, n=4) post-follicular ablation. The residual effect (T Residual, n=5) was evaluated in one subsequent, nonstimulated OPU cycle conducted 1 week later (9 days after last FSH administration). For plasma AMH, concentration was measured using ELISA kit (cat. no. AL114, AnshLabs; sensitivity 1.0 pgmL−1, interassay variation <5%, additional validation conducted with serial dilutions of buffalo plasma, r=0.98 against standard curve). Continuous data were analysed with ANOVA and Tukey's post hoc comparisons; categorical data were analysed using Chi-squared test. Pearson coefficient was used to evaluate correlation between AMH, follicular reserve, and invitro performance. Superovulatory treatment resulted in a higher number of follicles and good quality oocytes (grades 1 and 2) per donor, irrespective of OPU interval (T7 FSH: 3.0±2.55 and T14 FSH: 1.5±0.5 vs. T0 Control: 0.45±0.38 and T Residual: 0.4±0.4; P<0.05). There were more cleaved embryos (T7 FSH: 26%, T14 FSH; 30% vs. T0 Control: 7% and T Residual: 20%) and blastocysts (T7 FSH: 21%, T14 FSH; 30% vs. T0 Control: 5% and T Residual: 0; P<0.05) from FSH-treated donors. There was a strong correlation (r=0.9439) between follicle count and plasma AMH, and weaker relationships for oocytes recovered (r=0.3975), cleaved embryos (r=0.2317), or blastocysts (r=0.0106). Results indicate that FSH treatment with a 48-h coasting period results in significant improvements in follicular population, oocyte quality, and embryonic development; these effects are observed independently of the interval between initial follicular ablation and OPU. No residual effects of FSH treatment were observed, indicating that the beneficial effects of superstimulation are not carried over to the following OPU cycle. In conclusion, a strong correlation coefficient was established between initial plasma AMH level and follicular response to FSH treatment, indicating that it may be used as a selection criterion for animals likely to exhibit a good response to superstimulatory treatment.
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Abouhefnawy, H. A., and N. A. Wani. "76 Effect of season on the superstimulation response, embryo quality, and pregnancy establishment in dromedary camel (Camelus dromedarius)." Reproduction, Fertility and Development 33, no. 2 (2021): 146. http://dx.doi.org/10.1071/rdv33n2ab76.
Full textAbstract:
The present study was conducted to study the effect of season on the superstimulation response, embryo quality and pregnancy establishment in the dromedary camels. Two donors each were superstimulated in the month of June 2019 (non-breeding season) and January 2019 (breeding season) by a combination of 2500IU of equine chorionic gonadotrophin (eCG, Folligon; Intervet), given as a single intramuscular injection on Day 1 of the treatment protocol, and 400mg of pFSH (Folltropin; Bioniche) injected twice daily in declining doses of 2×80mg, 2×60mg, 2×40mg, and 2×20mg over 4 days, also beginning on Day 1. They were scanned by rectal ultrasonography and follicles counted before mating with a fertile bull on Day 11 after the start of treatment. An injection of 20µg of gonadotrophhin-releasing hormone (Receptal, Intervet) was given immediately after mating to induce ovulation. The number of corpora lutea (CL) were counted, and the embryos collected by the non-surgical method on Day 7 after ovulation. All embryos were washed and graded before they were transferred individually into the left uterine horn of recipient animals, synchronized to be on Day 6 after ovulation. All recipients were screened by ultrasonography on Day 22, Day 60, and Day 90 for pregnancies. The data were analysed using a two-sample t-test (Minitab statistical software, Minitab Ltd.). No difference was observed in the mean number of follicles developed (20.5 vs. 18.5) between the donors in breeding and non-breeding season, respectively. The percent mean±s.e.m. of ovulations as observed by counting the CL in nonbreeding season (80.7±3.4) were not different from those in breeding season (85.4±3.7). A total number of 16 embryos collected during non-breeding season and 20 embryos in breeding season were transferred to recipients. The percent mean±s.e.m. of pregnancies established on Day 22 (56.6±3.4 vs. 67.5±7.5) and on Day 90 (47.2±2.8 vs. 60±0.0) were similar in both groups. These observations indicate that embryo transfer technology can be applied to camels during the summer months, which is considered the non-breeding season, and pregnancies can be established at par with breeding season. In conclusion, to the best of our knowledge, this is the first study in camels wherein we have demonstrated that donor camels can be superstimulated, embryos can be collected, recipient animals can be synchronized, and pregnancies can be established in the summer (non-breeding) months. The embryos collected from elite donors can be transferred to synchronized recipients to establish pregnancies or preserved for future use. Some owners do not want calves to be born in summer months, so protocols need to be established for storing these embryos for a few months before their transfer to recipients, to have the calves delivered in the desired winter months.
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Abouhefnawy, H. A., and N. A. Wani. "76 Effect of season on the superstimulation response, embryo quality, and pregnancy establishment in dromedary camel (Camelus dromedarius)." Reproduction, Fertility and Development 33, no. 2 (2021): 146. http://dx.doi.org/10.1071/rdv33n2ab76.
Full textAbstract:
The present study was conducted to study the effect of season on the superstimulation response, embryo quality and pregnancy establishment in the dromedary camels. Two donors each were superstimulated in the month of June 2019 (non-breeding season) and January 2019 (breeding season) by a combination of 2500IU of equine chorionic gonadotrophin (eCG, Folligon; Intervet), given as a single intramuscular injection on Day 1 of the treatment protocol, and 400mg of pFSH (Folltropin; Bioniche) injected twice daily in declining doses of 2×80mg, 2×60mg, 2×40mg, and 2×20mg over 4 days, also beginning on Day 1. They were scanned by rectal ultrasonography and follicles counted before mating with a fertile bull on Day 11 after the start of treatment. An injection of 20µg of gonadotrophhin-releasing hormone (Receptal, Intervet) was given immediately after mating to induce ovulation. The number of corpora lutea (CL) were counted, and the embryos collected by the non-surgical method on Day 7 after ovulation. All embryos were washed and graded before they were transferred individually into the left uterine horn of recipient animals, synchronized to be on Day 6 after ovulation. All recipients were screened by ultrasonography on Day 22, Day 60, and Day 90 for pregnancies. The data were analysed using a two-sample t-test (Minitab statistical software, Minitab Ltd.). No difference was observed in the mean number of follicles developed (20.5 vs. 18.5) between the donors in breeding and non-breeding season, respectively. The percent mean±s.e.m. of ovulations as observed by counting the CL in nonbreeding season (80.7±3.4) were not different from those in breeding season (85.4±3.7). A total number of 16 embryos collected during non-breeding season and 20 embryos in breeding season were transferred to recipients. The percent mean±s.e.m. of pregnancies established on Day 22 (56.6±3.4 vs. 67.5±7.5) and on Day 90 (47.2±2.8 vs. 60±0.0) were similar in both groups. These observations indicate that embryo transfer technology can be applied to camels during the summer months, which is considered the non-breeding season, and pregnancies can be established at par with breeding season. In conclusion, to the best of our knowledge, this is the first study in camels wherein we have demonstrated that donor camels can be superstimulated, embryos can be collected, recipient animals can be synchronized, and pregnancies can be established in the summer (non-breeding) months. The embryos collected from elite donors can be transferred to synchronized recipients to establish pregnancies or preserved for future use. Some owners do not want calves to be born in summer months, so protocols need to be established for storing these embryos for a few months before their transfer to recipients, to have the calves delivered in the desired winter months.
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Satrapa, R. A., E. M. Razza, A. G. Pupulim, A. C. S. Castilho, B. Loureiro, J. S. Ticianelli, M. Machado, et al. "214 EFFECT OF OVARIAN SUPERSTIMULATION ON EXPRESSION OF GENES ASSOCIATED WITH THE OOCYTE DEVELOPMENTAL COMPETENCE OF NELORE COWS." Reproduction, Fertility and Development 25, no. 1 (2013): 255. http://dx.doi.org/10.1071/rdv25n1ab214.
Full textAbstract:
The P36 protocol has contributed to the genetic improvement of Brazilian herd through its successful use in embryo transfer programs. We aimed to investigate the effect of P36 protocol on embryo yield and mRNA expression of genes correlated with the competence of cumulus–oocyte complex (COC): receptors of FSH (FSHR), EGF (EGFR), and pentraxin 3 (PTX3) in cumulus cells; receptors of LH (LHR) and angiotensin 2 (AT2) in granulosa cells; and GDF9, BMP15, and histone H2A (H2A) in oocytes. Multiparous Nelore cows were allocated in control and P36 groups. Control group (non-superovulated, n = 15) received a progesterone intravaginal device (P4, 1.0 g, Primer®, Tecnopec, Sao Paulo, Brazil) and 2.5 mg of oestradiol benzoate (EB, IM, BER-BE®, Syntex, Buenos Aires, Argentina) at a random day of the oestrous cycle (Day 0). A PGF2α analogue (150 mg d-cloprostenol, IM, Prolise®, RARS SRL) was administered (Day 8) and Primer® was removed. The P36 group (n = 10) received a Primer® and 2.0 mg of EB (Day 0). The FSH treatment (160 mg Folltropin®, Bioniche Animal Health, Ontario, Canada) was initiated at decreasing doses: 40, 30, 20, and 10% of the total dose twice daily for 4 days (Day 5). The PGF2α analogue was administered (Day 8) and after 36 h primer was removed. Animal slaughter to ovary collection was performed 12 h after Primer® removal (Day 9). Some of the oocytes were matured (TCM199), fertilized with Nelore semen (n = 6), and cultured (SOF-synthetic oviduct fluid) to the blastocyst stage. Embryos were removed from culture (Day 6), allocated in 5 pools with 5 embryos in each group, and subjected to RNA extraction. Remaining oocytes were denuded from cumulus and zona pellucida (vortex and Protease®, Sigma-Aldrich, St. Louis, MO, USA). Pools of 20 oocytes and of their respective cumulus cells (n = 6 pools; control group and n = 4 pools, P36 group) were subjected to RNA extraction (RNeasy kit, Qiagen, Valencia, CA, USA). Gene expression was performed by real-time RT-PCR using oligo-dT in reverse transcription and bovine-specific primers. Expression of cyclophilin A was used as endogenous control. Change to developmental rates to the blastocyst stage and transcript abundance were compared by t-test and significance was considered when P < 0.05. Blastocyst rates were also similar (P > 0.05) in groups P36 (40/99; 40%) and control (16/43; 37%). Expression of H2A, EGFR, FSHR, and PTX3 in cumulus cells did not differ (P > 0.05) among treatment groups. The expression of GDF9 and BMP15 in cumulus cells was higher (P < 0.05) in the P36 group, but in oocytes these transcripts were more expressed in the control group (P < 0.05). Although important genes (GDF9 and BMP15) were less expressed in oocytes from superstimulated cows, the maintenance of H2A in oocytes, as well as PTX3, EGFR, and FSHR, and the increases in GDF9 and BMP15 expression in cumulus cells do not seem to affect oocyte competence due to the similar embryo yield of both groups. Supported by FAPESP.
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Dias, F. C. F., D. Dadarwal, M. Honparkhe, G. P. Adams, R. J. Mapletoft, and J. Singh. "313 EFFECT OF DURATION OF THE GROWING PHASE OF OVULATORY FOLLICLES IN SUPERSTIMULATED HEIFERS ON OOCYTE COMPETENCE AFTER IN VITRO FERTILIZATION." Reproduction, Fertility and Development 25, no. 1 (2013): 303. http://dx.doi.org/10.1071/rdv25n1ab313.
Full textAbstract:
We tested the hypotheses that extending the duration of follicular growth by superstimulation increases oocyte competence, and that FSH starvation at the end of superstimulatory treatment decreases oocyte competence. Heifers were allocated randomly to short FSH duration (n = 8), FSH starvation (n = 8), or long FSH duration (n = 8) groups. Five to 8 days after ovulation, transvaginal ultrasound-guided follicle ablation was done to synchronize follicle wave emergence, and a progesterone-releasing device (CIDR; Pfizer Animal Health, New York, NY, USA) was placed intravaginally. Short FSH and FSH starvation groups were given 8 doses of FSH (Folltropin-V; Bioniche Animal Health Inc., Belleville, ON, Canada) IM, whereas the long FSH group was given 14 doses of FSH at 12-h intervals, starting from the day of wave emergence (Day 0). Prostaglandin F2α (PGF) was administered twice, 12 h apart, on Day 3 in the short FSH group and on Day 6 in the other 2 groups. In all heifers, the CIDR was removed at the time of the second PGF treatment; pLH (Lutropin-V; Bioniche Animal Health Inc.) was given IM 24 h after CIDR removal, and cumulus–oocyte complexes (COC) were collected 24 h after pLH treatment. The COC were matured in vitro (6 h) and fertilized (IVF), and the embryos were cultured for 10 days. At 12 h after pLH, the long FSH group had a greater number of ≥9 mm follicles than the FSH starvation and short FSH groups (25.4 ± 5.3 v. 11.0 ± 2.1 and 10.6 ± 2.3, respectively; P < 0.03). The long FSH group also had more expanded COC than the FSH starvation group (P < 0.001), but did not differ from the short FSH group (93, 54, and 74%, respectively). The FSH starvation group had a greater proportion (P < 0.0001) of partially expanded COC (32%) and poor quality oocytes (70%) than did the long (1 and 33%) and short (4 and 45%) FSH groups; oocyte quality did not differ between long and short FSH groups. At 48 h after IVF, the cleavage rate was lower in the FSH starvation group compared with the short and long FSH groups (35, 54, and 56%, respectively; P = 0.003). After 9 days in culture, embryo development (morula + blastocyst) in the FSH starvation group was lower than that in the long FSH group, (18 v. 37%; P = 0.04), but did not differ from that in the short FSH group (25%). After removal of the data of one heifer in the FSH starvation group that produced 52% of total embryos in that group (outlier), the Day 9 blastocyst rate was lower in the FSH starvation group than in the short and long FSH groups (2% v. 14 and 21%, respectively; P = 0.02). In conclusion, extending the standard superstimulation protocol by 3 days enhanced ovarian response to FSH treatment, but did not improve oocyte competence, whereas a period of FSH starvation after FSH treatment compromised oocyte quality and embryo development.
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Bülbül, B., M. Kırbaş, Ş. Dursun, and M. Köse. "Superovulation in cows synchronized with two different progesterone+oestradiol protocols." Archives Animal Breeding 56, no. 1 (October 10, 2013): 160–68. http://dx.doi.org/10.7482/0003-9438-56-015.
Full textAbstract:
Abstract. A total of 26 Brown Swiss cows were used to compare the synchronization and superovulatory response of follicle stimulating hormone treated cows that were synchronized with progesterone+oestradiol valerate or benzoate. Control cows (n = 8) were superstimulated with follicle stimulating hormone using twice daily injections with decreasing doses from day 10–13 after determined reference oestrus. Cows in treatment groups were received either ear implant (n = 9) containing norgestomet+oestradiol valerate or progesterone releasing intravaginal device (n = 9) containing progesterone+oestradiol benzoate, at random stage of the oestrus cycle, for 9 days. Seven days after the implant and progesterone releasing intravaginal device insertion, follicle stimulating hormone was injected as described in the control group. There was no significant difference between the groups for superovulation responses. In conclusion, both protocols synchronized the oestrus cycle in follicle stimulating hormone treated cows and, any of the protocols evaluated in this study can be used as a pretreatment for superstimulation started on the seventh day of the implant or progesterone releasing intravaginal device insertion in Brown Swiss cows.
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Berland, M. A., A. von Baer, V. Parraguez, P. Morales, G. P. Adams, M. E. Silva, J. Ruiz, and M. H. Ratto. "263 IN VITRO FERTILIZATION AND EMBRYO DEVELOPMENT OF CUMULUS - OOCYTE COMPLEXES COLLECTED BY ULTRASOUND-GUIDED FOLLICULAR ASPIRATION IN LLAMAS TREATED WITH GONADOTROPIN." Reproduction, Fertility and Development 22, no. 1 (2010): 288. http://dx.doi.org/10.1071/rdv22n1ab263.
Full textAbstract:
We have previously documented that both FSH and eCG are equally effective in inducing ovarian superstimulation in llamas, resulting in the recovery of a high number of expanded COC suitable for in vitro fertilization (Ratto et al. 2005 Theriogenology 63, 2445-2457). The objective of the study was to evaluate the ovarian response, morphology, and competence of COC collected by ultrasound-guided follicular aspiration in llamas treated with FSH or eCG. Llamas were assigned randomly into 2 groups (n = 16 per group) and treated for 48 h after follicle ablation with (1)25 mg of FSH (Folltropin, Bioniche Animal Health Canada Inc., Belleville, Canada) i.m. twice daily for 4 d; or (2) 1000IU of eCG (Novormon, Bioniche Animal Health Canada) as a single i.m. dose. The starting of gonadotropin treatment was considered Day 0. Both groups were given an i.m. dose of 5 mg of Armour Standard LH (Lutropin, Bioniche Animal Health Canada) on Day 6, and COC were collected by transvaginal ultrasound follicle aspiration of all follicles ≥7 mm on Day 7. The ovarian response was assessed by transrectal ultrasonography using a 7.5-MHz linear-array transducer (Aloka SSD-500, Clinics, Santiago, Chile) immediately before oocyte collection at 24 to 26 h after LH treatment in both groups. The COC were classified as expanded, compact, denuded, or degenerated. Expanded COC collected from FSH- (n = 147) and eCG-treated llamas (n = 141) were fertilized in vitro using epididymal sperm as previously described (Ratto et al. 2006 Anim. Reprod. Sci. 97, 246-257). Gametes were co-incubated at 38.5°C in air with 5% CO2 and high humidity for 18 h. After in vitro fertilization, presumptive zygotes were co-culture in SOF medium supplemented with 0.6% of BSA with llama granulosa cells at 39°C, 5% CO2, 5% O2, and 90% N2 for 7 days. Embryo development was evaluated on Days 2, 5, and 7 of in vitro culture (Day 0 = IVF). Data were analyzed by Student’s t-test or Fisher’s exact test and presented as mean ± SEM. The FSH and eCG treatment groups did not differ with respect to the number of follicles ≥7 mm at the time of COC collection (16.0 ± 2.7 v. 14.0 ± 1.9; P = 0.5), the number of COC collected (11.5 ± 1.9 v. 9.7 ± 1.2; P = 0.4), or the collection rate per follicle aspirated (77.0 v. 71.5%; P = 0.2). No difference was detected between FSH and eCG-treated llamas in the number of expanded COCs (9.8 ± 1.4 v. 9.4 ± 1.2; P = 0.8). The percentage of presumptive zygotes to develop into 2 to 8 cells on Day 2 (65.3 v. 63.1), morulas on Day 5 (46.2 v. 42.5), and blastocyst stage on Day 7 (23.1 v. 20.5) did not differ (P > 0.05) between FSH and eCG-treated llamas, respectively. In conclusion, FSH and eCG treatments were equally effective for ovarian superstimulation and oocyte collection. The recovery of a high number of expanded COC can be used directly for in vitro fertilization and their competence is not affected by gonadotropin treatment. The study was supported by Convenio Desempeño en Investigacion (2007-DGI-CDA-04), Universidad Catolica de Temuco.
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Hayden, C., R. V. Sala, D. C. Pereira, L. C. Carrenho-Sala, M. Fosado, D. Moreno, J. F. Moreno, and A. Garcia-Guerra. "71 Effect of follicle-stimulating hormone dose and circulating progesterone before ovum pickup and invitro embryo production in pregnant Holstein heifers." Reproduction, Fertility and Development 33, no. 2 (2021): 143. http://dx.doi.org/10.1071/rdv33n2ab71.
Full textAbstract:
Superstimulation with exogenous FSH before ovum pickup for invitro embryo production (IVP) improves embryo production. However, there is limited information on different FSH dose regimens and the effect of progesterone (P4) concentrations on IVP. Thus, the objective of the present study was to evaluate the effect of FSH dose and circulating P4 on ovarian superstimulation responses, before ovum pickup (OPU) on IVP in pregnant heifers. Pregnant Holstein heifers (n=47) 19.5±0.3 months of age with moderate body condition score (3.3±0.03; scale 1 to 5) were randomly assigned to 2 treatment groups, p-FSH (Folltropin-V, Vetoquinol) 160mg (FSH160) or p-FSH 300mg (FSH300), in a crossover design. Two sessions of OPU were performed on each animal at 48 and 62 days of gestation, with a washout interval between sessions of 14 days. Dominant follicle removal (DFR) was utilised to synchronize follicular wave emergence, and FSH treatments were initiated 36h later. Heifers in both groups received a total of 4 FSH injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3, and 32.0mg) or FSH300 (90.0, 80.0, 70.0, and 60.0mg). All heifers underwent OPU 40h after the last FSH treatment. Blood samples were collected for P4 quantitation at OPU and number of small (<6mm), medium (6–10mm), and large (>10mm) follicles were determined before OPU. Cumulus–oocyte complexes (COCs) recovered were processed and invitro embryo production was performed using standard procedures. Differences between treatment groups were evaluated using generalized linear mixed models, and data are presented as mean±s.e.m. (Table 1). Heifers treated with 300mg of p-FSH had a greater (P<0.05) number of medium-size follicles, total number of follicles, total COCs recovered, number of viable COCs, and number of blastocysts produced. In addition, heifers treated with 300mg of p-FSH had a greater cleavage rate (P=0.03). Circulating P4 was on average 10.1±0.4ng mL−1 and did not differ between gestation days (P=0.12). Furthermore, circulating P4 did not affect (P>0.10) any of the outcomes evaluated, nor was there an FSH dose by circulating P4 interaction (P>0.05). In conclusion, increasing the dose of p-FSH before OPU from 160 to 300mg resulted in an increase in the total number of follicles, number of oocytes, and number of blastocysts produced in pregnant donors without changes in blastocyst rate, regardless of progesterone concentrations at the time of OPU. Table 1. Ovum pickup and invitro embryo production in pregnant heifers treated with different doses of p-FSH Item FSH160 FSH300 P-value Small follicles (n) 5.0±0.6 3.5±0.4 0.02 Medium follicles (n) 14.4±0.8 18.3±0.9 0.0003 Large follicles (n) 2.9±0.3 2.5±0.3 0.28 Total follicles (n) 22.3±0.7 24.3±0.9 0.04 Total oocytes (n) 12.8±0.7 14.7±1.0 0.02 Recovery rate (%) 56.2±2.1 59.7±2.7 0.20 Viable oocytes (n) 11.4±0.7 13.3±1.0 0.02 Viable oocytes (%) 89.9±1.7 88.0±2.4 0.22 Cleavage rate (%) 63.8±4.3 71.5±3.7 0.03 Cleaved oocytes (n) 7.2±0.7 9.3±0.9 0.14 Blastocysts (n) 2.7±0.4 3.6±0.7 0.02 Blastocyst rate (%) 22.0±3.3 22.4±3.6 0.74
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Hayden, C., R. V. Sala, D. C. Pereira, L. C. Carrenho-Sala, M. Fosado, D. Moreno, J. F. Moreno, and A. Garcia-Guerra. "71 Effect of follicle-stimulating hormone dose and circulating progesterone before ovum pickup and invitro embryo production in pregnant Holstein heifers." Reproduction, Fertility and Development 33, no. 2 (2021): 143. http://dx.doi.org/10.1071/rdv33n2ab71.
Full textAbstract:
Superstimulation with exogenous FSH before ovum pickup for invitro embryo production (IVP) improves embryo production. However, there is limited information on different FSH dose regimens and the effect of progesterone (P4) concentrations on IVP. Thus, the objective of the present study was to evaluate the effect of FSH dose and circulating P4 on ovarian superstimulation responses, before ovum pickup (OPU) on IVP in pregnant heifers. Pregnant Holstein heifers (n=47) 19.5±0.3 months of age with moderate body condition score (3.3±0.03; scale 1 to 5) were randomly assigned to 2 treatment groups, p-FSH (Folltropin-V, Vetoquinol) 160mg (FSH160) or p-FSH 300mg (FSH300), in a crossover design. Two sessions of OPU were performed on each animal at 48 and 62 days of gestation, with a washout interval between sessions of 14 days. Dominant follicle removal (DFR) was utilised to synchronize follicular wave emergence, and FSH treatments were initiated 36h later. Heifers in both groups received a total of 4 FSH injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3, and 32.0mg) or FSH300 (90.0, 80.0, 70.0, and 60.0mg). All heifers underwent OPU 40h after the last FSH treatment. Blood samples were collected for P4 quantitation at OPU and number of small (<6mm), medium (6–10mm), and large (>10mm) follicles were determined before OPU. Cumulus–oocyte complexes (COCs) recovered were processed and invitro embryo production was performed using standard procedures. Differences between treatment groups were evaluated using generalized linear mixed models, and data are presented as mean±s.e.m. (Table 1). Heifers treated with 300mg of p-FSH had a greater (P<0.05) number of medium-size follicles, total number of follicles, total COCs recovered, number of viable COCs, and number of blastocysts produced. In addition, heifers treated with 300mg of p-FSH had a greater cleavage rate (P=0.03). Circulating P4 was on average 10.1±0.4ng mL−1 and did not differ between gestation days (P=0.12). Furthermore, circulating P4 did not affect (P>0.10) any of the outcomes evaluated, nor was there an FSH dose by circulating P4 interaction (P>0.05). In conclusion, increasing the dose of p-FSH before OPU from 160 to 300mg resulted in an increase in the total number of follicles, number of oocytes, and number of blastocysts produced in pregnant donors without changes in blastocyst rate, regardless of progesterone concentrations at the time of OPU. Table 1. Ovum pickup and invitro embryo production in pregnant heifers treated with different doses of p-FSH Item FSH160 FSH300 P-value Small follicles (n) 5.0±0.6 3.5±0.4 0.02 Medium follicles (n) 14.4±0.8 18.3±0.9 0.0003 Large follicles (n) 2.9±0.3 2.5±0.3 0.28 Total follicles (n) 22.3±0.7 24.3±0.9 0.04 Total oocytes (n) 12.8±0.7 14.7±1.0 0.02 Recovery rate (%) 56.2±2.1 59.7±2.7 0.20 Viable oocytes (n) 11.4±0.7 13.3±1.0 0.02 Viable oocytes (%) 89.9±1.7 88.0±2.4 0.22 Cleavage rate (%) 63.8±4.3 71.5±3.7 0.03 Cleaved oocytes (n) 7.2±0.7 9.3±0.9 0.14 Blastocysts (n) 2.7±0.4 3.6±0.7 0.02 Blastocyst rate (%) 22.0±3.3 22.4±3.6 0.74
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Hinshaw, R. H., M. L. Switzer, R. J. Mapletoft, and G. A. Bó. "351 A COMPARISON OF 2 APPROACHES FOR THE USE OF GnRH TO SYNCHRONIZE FOLLICLE WAVE EMERGENCE FOR SUPEROVULATION." Reproduction, Fertility and Development 27, no. 1 (2015): 263. http://dx.doi.org/10.1071/rdv27n1ab351.
Full textAbstract:
Although oestradiol has been used successfully to synchronize follicle wave emergence for superovulation, it cannot be used in many countries. Attention has turned to alternatives, including the use of GnRH to induce ovulation of a dominant follicle, which will be followed by emergence of a new follicular wave in 1 to 2 days. However, GnRH synchronizes follicular wave emergence only when it induces ovulation and administration of GnRH at random stages of the oestrous cycle results in ovulation in less than 60% of animals. The objective of the study was to compare superovulatory response and ova/embryo production following synchronization of follicle wave emergence for superovulation with GnRH administered 2 days after insertion of a progestin device, with a protocol in which GnRH is administered 7 days after administration of prostaglandin F2α and a progestin device. Beef donors of various breeds were placed at random into 1 of 2 groups and superstimulated by replicate so that one cow in each group had ova/embryos collected on the same day. Sixty-six superstimulations were performed in this study; 26 were performed in 13 donors that were superstimulated twice in a crossover design, and 40 donors were superstimulated once (i.e. 20 donors in each treatment group). Cows in group 1 received CIDR devices (Zoetis Animal Health, USA) on Day –2 and 100 μg of GnRH (Cystorelin, Merial USA) on Day 0; FSH treatments were initiated on Day 2 with 288 mg of Folltropin-V (Vetoquinol, Canada) given in twice-daily decreasing doses for 4 days. Prostaglandin F2α (PGF; 35 mg dinoprost, Lutalyse, Zoetis) was given with the last 2 injections of Folltropin-V and CIDR were removed with the last Folltropin-V administration (i.e. Day 6). Donors received a second GnRH at the onset of oestrus and were AI 8 and 20 h later. Donors that were still in standing oestrus at the second AI were AI again at 30 h. Ova/embryo collections were done on Day 14 and embryos were classified according to the IETS manual. Cows in group 2 received an injection of PGF and a CIDR on Day –7 and 100 μg of GnRH on Day 0; FSH treatments were initiated on Day 2 and the remainder of the treatment protocol was as in group 1. Data (total ova/embryos collected and transferable embryos) were analysed by ANOVA for mixed models, using treatment as a fixed variable and cow (i.d.) as a random variable. The group 1 cows produced a mean (± s.e.m.) of 18.6 ± 1.9 total ova/embryos of which 12.7 ± 1.5 were of transferable quality (7.2 ± 1.3 grade 1). Cows in group 2 produced a mean (± s.e.m.) of 19.5 ± 1.7 total ova/embryos, of which 14.8 ± 1.5 were of transferable quality (8.9 ± 1.2 grade 1). Although 2 more transferable embryos were obtained in group 2, differences were not significant (P > 0.3). At the same time as this experiment was done, 214 other cows were superstimulated in this practice, yielding an average of 7.9 transferable embryos per donor. Results suggest that both approaches are efficacious for the superstimulation of beef cows.Special thanks to Vetoquinol/Bioniche Animal Health, USA for support.
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Ratchamak, Ruthaiporn, Thanaporn Ratsiri, Rujira Chumchai, Wuttigrai Boonkum, and Vibuntita Chankitisakul. "Relationship of the Temperature-Humidity Index (THI) with Ovarian Responses and Embryo Production in Superovulated Thai-Holstein Crossbreds under Tropical Climate Conditions." Veterinary Sciences 8, no. 11 (November 8, 2021): 270. http://dx.doi.org/10.3390/vetsci8110270.
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Heat stress strongly negatively affects reproductive traits in dairy cattle. The purpose of the present study was to investigate the effect of heat stress in superstimulated Thai-Holstein crossbreds under tropical climate conditions. Data included 75 records from 12 superovulated Thai-Holstein crossbreds between 2018 and 2020. Cows were superstimulated with conventional treatment. The mean temperature-humidity index (THI) was evaluated for three data collection periods: during the 9, 21 and 42-day periods before the insemination day to determine the period in which THI mostly affected superstimulation responses. The THI levels/thresholds were determined and interpreted together with the superovulatory response. A significantly negative correlation was obtained for the THI during the period 9 days before insemination. Negative effects on the number of large follicles and corpora lutea began at a THI of 72 and were apparently severe after a THI of 77, similar to the ovulation rate, fertilized ova and transferable embryos (p < 0.05). Meanwhile, more degenerated embryos were found with THI values (p < 0.05). The superovulatory response in Thai-Holstein crossbreds under tropical climate conditions is highly affected by heat stress starting at a THI of 72 and becomes more severe at a THI higher than 77.
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Caunce, S. L., A. R. T. Krause, F. C. F. Dias, G. P. Adams, and J. Singh. "4 Relationship Between Ovarian Vascularity, Cumulus–Oocyte Morphology and Luteal Development in Four-Month-Old Calves After FSH Stimulation." Reproduction, Fertility and Development 30, no. 1 (2018): 141. http://dx.doi.org/10.1071/rdv30n1ab4.
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The objectives of this study were to determine the effect of LH on the blood flow to the ovaries of 4-month-old calves after 2 FSH stimulation protocols, and to examine the relationship between ovarian vascularity after superstimulation to the morphology of the cumulus–oocyte complexes (COC) and luteal function. We hypothesise that ovarian vascularity (detected by 3-dimensional (3D) analysis of Doppler ultrasound cineloops) will increase in response to LH, and the magnitude of change in vascularity would be predictive of (1) a greater proportion of expanded COC, (2) greater development of luteal tissue volume and vascularity at 3 and 7 days after follicle aspiration, and (3) higher levels of plasma progesterone. Ovarian superstimulation was initiated at the beginning of an induced follicular wave in 4-month-old beef calves (n = 16), and beef cattle >16 months of age (control group, adults; n = 8) using either a traditional 4-day or an extended 7-day FSH protocol (n = 8 calves and n = 4 controls per group). Power Doppler ultrasound cineloops were recorded immediately before (i.e. 12 h after the last FSH treatment) and 24 h after LH treatment (before ultrasound-guided follicular aspiration for oocyte collection) to assess ovarian vascularity, and 3 and 7 days after follicular aspiration to assess luteal tissue volume and vascularity. Video segments were analysed in Fiji and Imaris software to obtain the 3D ovarian vascularity index (ratio of blood flow volume to tissue volume). The ovarian vascularity index tended to increase >1.7-fold in response to exogenous LH in both prepubertal calves (pre-LH 1.5 ± 0.4% v. post-LH 2.6 ± 0.7%; P = 0.08) and adult cattle (pre-LH 2.2 ± 0.6% v. post-LH 4.7 ± 0.9%; P = 0.07). Calves with a recovery of >75% of expanded COC had a higher ovarian vascularity index (10.7 ± 2.6% v. 4.8 ± 1.6%; P = 0.06) and luteal vascularity index (15.7 ± 4.5% v. 5.7 ± 2.1%; P < 0.05) 7 days after aspiration than those with <75% expanded COC. Calves in the 7-day FSH protocol had >10-fold higher concentration of plasma progesterone on Day 3 (12.7 ± 7.3 ng mL−1 v. 1.2 ± 0.4 ng mL−1; P < 0.05) and Day 5 (50.6 ± 28.0 ng mL−1 v. 4.5 ± 1.0 ng mL−1; P < 0.05), and ~2-fold higher luteal vascularity index at 7 days after follicle aspiration (13.7 ± 4.6% v. 7.7 ± 2.8%; P < 0.05) than calves in the 4-day FSH protocol, whereas no difference (P > 0.05) was found in control (adult) animals. In conclusion, there was an increase in ovarian vascularity resulting from LH treatment in prepubertal calves and adult cattle. A greater proportion of expansion of COC at 24 h after LH treatment (an indicator of follicular maturation) was related to higher ovarian and luteal vascularity on Day 7 after collection in prepubertal calves, but not in adults. Luteal vascularity on Day 3 was reflective of plasma progesterone concentration, and prepubertal calves in the 7-day FSH protocol had greater plasma progesterone than calves in the 4-day FSH protocol. The use of FSH in calves allows a greater number of follicles for oocyte collection as it does in adult cattle. Research was supported by an NSERC grant.
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Hegedüsová, Z., R. Holásek, M. Slezáková, A. Dufek, and J. Kubica. "141 EFFECT OF ENVIRONMENTAL TEMPERATURE ON EMBRYO PRODUCTION AND CONCEPTION RATES IN BEEF AND DAIRY CATTLE." Reproduction, Fertility and Development 21, no. 1 (2009): 170. http://dx.doi.org/10.1071/rdv21n1ab141.
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The aim of this study was to evaluate the effect of environmental temperature at the time of embryo collection and transfer on embryo quality and conception rates after transfer of fresh and frozen embryos. Purebred dairy (Holstein, Czech spotted cattle, n = 174) and beef (Charolais, Blonde d’Aquitaine, Piedmontese, Hereford, Limousin and Simmental; n = 72) donors were collected during 2005 to 2007. Donor cows received one application of PGF2α and superstimulation was initiated 9 to 11 days following estrus with pFSH (Folltropin-V; Bioniche Animal Health, Belleville, ON, Canada) given twice daily for 4 days. Prostaglandin2α was given on Day 3 of the superstimulation treatment. Donors were inseminated 3 times, with 1 unit of semen on Days 5 and 6 of treatment. Embryo recovery was carried out on Day 7 after insemination. The recipients were synchronized with PGF2α and embryos were transferred on Day 7 after estrus. The effect of temperature at the time of 246 embryo recoveries and 1338 transfers (fresh and frozen embryos) was analyzed using the GLM, assuming quasi-poisson and quasi-binomial error distribution by R software (www.r-project.org). We evaluated the total number of ova/embryos collected, embryo quality, and conception rates after transfer at the environmental temperatures shown below. Temperatures were divided into the following ranges: A) from –5 to 5°C; B) from 6 to 15°C; C) from 16 to 20°C; and D) over 20°C. The average temperature values were obtained from the Czech hydrometeorological institute and corresponded to the following seasons: winter (range A and B); spring (range B and C); summer (range C and D); fall (range C and B). Dairy cows were managed intensively in barns whereas beef cows were managed in barns during the winter and on pasture from April until October. In beef donors the mean number of total ova/embryos collected and transferable embryos were 9.1 ± 7.9 and 4.2 ± 4.5, respectively and no significant effect of temperature was detected (P > 0.05). In dairy donors the mean number of total ova/embryos collected and transferable embryos were 7.9 ± 6.0 and 4.9 ± 4.5, respectively and no significant effect of temperature was detected (P = 0.27). Conception rate was greatest (P < 0.001) in temperature range D (63.64%) compared with the other temperature ranges (A = 27.66%; B = 43.65%; C = 43.21%;) in beef cows and in the range C (65.31%) than in the other temperature ranges (A = 34.7%; B = 52.67%; D = 56.25%) in dairy cows. It was concluded that temperature at the time of embryo collection did not affect embryo production and quality. However, environmental temperature did have a significant effect on conception rates in both beef and dairy recipients. Nutritional status may also have contributed to the increased conception in beef cattle during warmer temperatures. However, the optimal period for conception in dairy cows was in temperature range from 16 to 20°C, because of the potential adverse effect of heat stress in high-yielding cows. Supported by MEYS CR MSM 2678846201, LA 330; NAZV 1B44034.
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Maclellan, L. J., T. R. Whyte, A. Murray, L. A. Fitzpatrick, C. R. Earl, W. J. Aspden, J. E. Kinder, et al. "Superstimulation of ovarian follicular growth with fsh, oocyte recovery, and embryo production from zebu (bos indicus) calves: Effects of treatment with a gnrh agonist or antagonist." Theriogenology 49, no. 7 (May 1998): 1317–29. http://dx.doi.org/10.1016/s0093-691x(98)00079-x.
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Kohram, H., V. Vahedi, S. Nasrollahi, and A. Farahavar. "Superovulation following follicular synchronization with GnRH at random stages of the oestrous cycle in heifers." Czech Journal of Animal Science 56, No. 1 (January 20, 2011): 7–14. http://dx.doi.org/10.17221/325/2009-cjas.
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The objective of this study was to develop a superovulatory program based on synchronization of follicular waves with GnRH which could be applied regardless of the stage of the oestrous cycle. 36 heifers were subjected to this experiment and GnRH (Cystorelin, 200 µg) was applied between Days 0 and 7 (n = 15), 8 and 12 (n = 8) or 13 and 20 (n = 13) of the oestrous cycle. Four days after GnRH treatment, all follicles ≥ 6 mm of heifers (n) were either punctured (n = 21) or left intact (n = 15). All heifers were superstimulated from Day 6 to Day 10 after GnRH treatment with 320 mg Folltropin-V. In parallel, 21 heifers were superstimulated in a conventional manner (Days 8 to 12) and were used as controls. The homogeneity of follicular inventories among Stage-groups occurred within 4 days of GnRH treatment for follicles ≥ 7 mm but only 2 days after follicular puncture for follicles 4 to 6 mm. In response to the follicular puncture, the mean number of follicles 4 to 6 mm increased in heifers of the punctured group (P < 0.01). Following the superstimulation, the follicular (P < 0.01) and ovulatory (P < 0.01) responses were higher in the punctured group than in the nonpunctured group. The in vivo production of transferable embryos in the punctured group was similar to that of the nonpunctured group but it was lower (P < 0.01) than in heifers of the control group. In conclusion, results from the present study indicate that regardless of the stage of the oestrous cycle, the homogeneity of follicular inventories following the follicular synchronization is obtained using GnRH treatment and follicular puncture. The in vivo production of embryos was severely compromised in the present study with heifers. Causes of such reduction in the in vivo production of embryos are still unknown.
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Chinarov, R. Y., G. N. Singina, V. Havlicek, N. P. Taradajnic, T. E. Taradajnic, and S. V. Pozyabin. "123 Efficiency of embryo production using ovum pickup oocytes recovered from stimulated and nonstimulated cows." Reproduction, Fertility and Development 33, no. 2 (2021): 169. http://dx.doi.org/10.1071/rdv33n2ab123.
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Recovery of oocytes from live animals through ultrasound-guided transvaginal follicular aspiration (ovum pickup, OPU) is an essential element of modern livestock development. To increase the number of OPU oocytes retrieved, hormonal stimulation is broadly applied; however, the results are ambiguous. The aim of the present study was to compare recovery rate, quality of oocytes, and efficiency of invitro production of cattle embryos using OPU oocytes derived from hormonal treated and untreated cows. The study was performed in Simmental heifers at the age of 17 to 23 months. The heifers in the first group (n=7) were previously synchronized using prostaglandin F2α (Estrumate, MSD Animal Health) and gonadotrophin-releasing hormone (GnRH; Receptal, MSD Animal Health) and then underwent superstimulation with Stimofol (Reprobiol SPRL, Belgium) in a total amount of 0.25mg of FSH and 0.05mg of LH in a treatment of 6 injections on 3 consecutive days starting on Day −4 before OPU. The heifers of the second group (n=2) underwent OPU once a week for 5 weeks (10 OPU sessions). Groups of recovered cumulus–oocyte complexes (COCs) were matured in TCM-199 containing 10% fetal calf serum, 1mM Na-pyruvate, 50μg mL−1 gentamycin, 10μg mL−1 FSH, and 10μg mL−1 LH for 24h, fertilized in Fert-TALP (Tyrode’s-albumin-lactate-pyruvate) for 18 to 20h, and cultured in CR1aa medium for 7 days. All steps of IVP were performed at 38.5°C with 5% CO2 and 90% humidity. The cleavage and blastocysts rates were assessed on Day 2 and 7, respectively. In addition, Day-7 blastocysts were fixed, and the total cell number was determined using 4′,6-diamidino-2-phenylindole (DAPI) staining. The data were analysed by ANOVA using SigmaStat software package. As expected, the number of aspirated follicles per session in the first group of cows was higher than that in the second group: 14.6±1.2 vs. 6.8±0.5 (P<0.05). In total, 54 and 42 COCs were collected, which corresponds to recovery rates of 54.5±7.7 and 61.3±4.9% for stimulated and nonstimulated cows, respectively. After the quality evaluation, 36 COCs of SS heifers (67.2±1.9%) and 31 COCs of non-SS heifers (75.3±5.0%) were selected for IVP. We did not observe a significant difference in cleavage rate between two groups: 77.8% in the SS group vs. 71.0% in the non-SS group, whereas the rate of blastocyst production was higher (P<0.05) for SS heifers (25.0%) compared with non-SS animals (16.1%). The total number of cells in Day 7 blastocysts was significantly (P<0.01) higher in the first group (89.8±1.3) compared with the second group (70.6±3.2). Thus, superstimulation of heifers before OPU increases the efficiency of oocyte retrieval by ultrasound-guided transvaginal follicular aspiration as well as the developmental competence of OPU-derived bovine oocytes invitro. These studies were performed under financial support of the Russian Science Foundation (project No. 19-16-00115) and the Ministry of Science and Higher Education of Russia.
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Barfield, J. P., and G. E. Seidel. "311 SUPEROVULATION OF NORTH AMERICAN BISON WITH TWO INJECTIONS OF FOLLICLE-STIMULATING HORMONE." Reproduction, Fertility and Development 25, no. 1 (2013): 302. http://dx.doi.org/10.1071/rdv25n1ab311.
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Few studies have examined superovulation of North American Bison. In cattle, ovarian superstimulation is usually achieved with 6 to 8 injections of FSH at half-day intervals. However, handling bison repeatedly stresses the animals, which could adversely affect their reproductive potential, as well as pose a risk of injury to the bison and handlers. To limit the number of times the bison were handled, we tested a two-injection superovulation scheme using sodium hyaluronate (MAP-5, Bioniche Animal Health Inc., Belleville, Ontario, Canada) in the FSH diluent, which serves to slow the absorption of FSH when given IM. We hypothesised that the two-injection superstimulation protocol would result in recovery of more embryos on average than a single-embryo recovery protocol. Although a traditional superovulation scheme with 6 to 8 injections of FSH would have been a better comparison, our goal was to handle the bison minimally. Eight female bison ranging in age from 5 to 11 years were used as embryo donors. For superovulation, females with a corpus luteum (CL) were given 25 mg of prostaglandin F2α (PGF, Lutalyse, Pfizer Animal Health, New York, NY, USA) IM followed by 266 mg of FSH (Folltropin V in MAP-5 diluent, i.m., Bioniche Animal Health Inc.) 12 days later (or 9 days after presumed oestrus 3 days post-PGF). Forty-eight hours after the first FSH injection, 134 mg of FSH IM and 25 mg of PGF IM were given. Two days later females were put in a pen with a bison bull for natural breeding. Seven days after assumed oestrus, embryos were recovered nonsurgically. Although the situation is not clear in bison, there is evidence in cattle that superovulated cycles influence embryo collections in subsequent cycles. Consequently, females were randomly assigned to a superovulation or single-embryo recovery treatment for each cycle; however, consecutive superovulation protocols were never conducted without a short oestrous cycle in between. Superovulated females were given PGF at embryo recovery after superovulation, followed by PGF 12 to 14 days later, and bred off the assumed oestrus of the second PGF injection. Embryos were collected from females 4 times (2 superovulation and 2 single embryo cycles, except one bison that was superovulated once). Data were analysed using a one-tailed t-test. Superovulation resulted in greater mean numbers of palpable CL (3.7; P < 0.001), embryos collected (1.8; P < 0.05), and transferable quality embryos (0.8; P < 0.05) compared with the single-embryo recovery protocol (mean palpable CL, 1.0; embryos collected, 0.5; transferable embryos, 0.2). Notably, the bison breeding season is July to September and occasionally animals breed in October; this experiment was conducted in October and November. Thirteen transferable embryos were nonsurgically transferred to recipients; 6 pregnancies were established, but 5 were resorbing by 2 months of gestation; 1 healthy calf was carried to term. Two injections of FSH with a long-acting diluent can be used to increase the number of embryos recovered from bison compared with a single-embryo recovery scheme.