Serum progesterone concentration and conception rate of beef cows supplemented with ground corn after a fixed-time artificial insemination protocol

Avaliaram-se os efeitos de diferentes níveis de ingestão de suplemento com milho moído finamente (MF) em vacas de corte, mantidas em pasto, após inseminação artificial em tempo fixo (IATF), sobre a concentração sérica de progesterona (P4) no dia 7, e sobre a concepção no dia 28 pós IATF. Trezentas e sessenta e quatro vacas Brangus, multíparas lactantes, tiveram as atividades folicular e luteal sincronizadas por tratamento com benzoato de estradiol (Estrogin; 2,0mg IM) e inserção de dispositivo intravaginal de P4 (CIDR) no dia -11, seguido por tratamento com PGF2 α (Lutalyse; 25mg IM) no dia - 4, retirada do CIDR e remoção temporária de bezerros no dia -2, e tratamento com GnRH (Fertagyl; 100 µ g IM), IATF e retorno dos bezerros no dia 0. No dia 0, as vacas foram aleatoriamente distribuídas para receber um dos quatro tratamentos: G1 -2kg/dia de MF do dia 0 ao dia 21; G2 -2kg/dia de MF do dia 0 ao dia 7, e 6kg/dia de MF do dia 8 ao dia 21; G3 -6kg/dia de MF do dia 0 ao dia 7, e 2kg/dia de MF do dia 8 ao dia 21; G4 -6kg/dia de MF do dia 0 ao dia 21. Amostras de sangue foram colhidas no dia 7, e o diagnóstico de gestação foi realizado por ultrassonografia no dia 28. As vacas suplementadas com 2kg/dia de MF apresentaram maior concentração sérica de P4 no dia 7 em relação às vacas suplementadas com 6kg/dia (1,58 vs. 1,28ng/mL; P<0,01, EPM=0,08). As vacas do G4 apresentaram maior taxa de concepção em relação às vacas do G1 (58,4 vs. 41,9%, respectivamente; P<0,05). O nível de consumo do suplemento energético após a IATF é negativamente associado às concentrações séricas de P4, porém positivamente associado à taxa de concepção em vacas de corte em pasto.

Ovarian response of Suffolk ewes to estrous synchronization using short-term protocol

The efficacy of estrus synchronization using short-term protocol was evaluated by ultrasound exams in Suffolk ewes during the pre-breeding season. The control Group (n = 12) was synchronized by treatment for 12 days with vaginal sponges impregnated with medroxyprogesterone acetate, and 400 IU eCG at sponge withdrawal. Experimental groups I, II and III kept the sponge in place for 4 days, and 100 µg of PGF2a was administered at sponge withdrawal. Additionally, Group I (n = 12) had 0.1 mg of estradiol benzoate (EB) administered during sponge placement and 50 µg of GnRH 48 hours after sponge removal. Group II (n = 6) had 35 mg of progesterone (P4) injected, and 0.1 mg of EB administered during sponge placement, 400 IU eCG at withdrawal and 48 hours after, 50 µg GnRH were administrated. Group III (n = 12) had 35 mg of P4 and 0.2 mg of EB administered at sponge placement, 400 IU eCG at withdrawal, and 50 µg of GnRH was administrated after 56 hours. Ovaries were monitored through ultrasound scanning. Concerning the first wave, no difference was detected between the control group and the experimental groups. However, the characteristics of ovulatory wave were significantly different between the groups. The duration of the follicular wave was shorter for Group III than for Group II. The follicle in Group I reached its maximum diameter before the Group II. The diameter of the follicle at the sponge withdrawal in the control group was larger than in Group I. After sponge withdrawal, the follicular growth rate was smaller in the control group than in Group III. The maximum diameter of the follicle in Group II was larger than in the other groups. The short-term protocol in which estrogen was used did not synchronize the emergence of the wave of follicular development.

Improvement of embryo production by the replacement of the last two doses of porcine follicle-stimulating hormone with equine chorionic gonadotropin in Sindhi donors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reuse of norgestomet implants in an eCG-based superovulation protocol administered to Nelore (Bos taurus indicus) cows

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Improvement in embryo recovery using double uterine flushing

The objective of the present study was to evaluate the effects of double uterine flushing on the recovery of embryos/ova in cattle. Two hundred and ten embryo recovery procedures were conducted using a double uterine flushing method, and the results were compared with 432 conventional single-flushing procedures. Cyclic Limousin (n = 403) and Guzera (n = 239) donor cows received an intravaginal progesterone releasing device and 2 mg of estradiol benzoate on Day 0. Between Days 5 and 9, donors received decreasing doses of FSH, which ranged from 200 to 300 IU (Bos indicus) and 300 to 500 IU (Bos taut-us). on the afternoon of Day 7, donors received an injection of 500 mu g cloprostenol and progesterone implants were removed 12 It later (morning of Day 8). Artificial insemination was performed between 14 and 26 h after first detection of behavioral estrus. Cows were randomly assigned to have embryos recovered by a double-flushing method (n = 210) or the conventional single-flushing procedure (n = 432). For the double-flushing procedure, after first flushing the whole uterus with 1 L of Dubelco's Phosphate Buffered Saline (DPBS), a Foley catheter was positioned in the uterine body to permit refilling of the uterus with fresh DPBS (80-150 mL). The catheter was closed with the plunger of a disposable 5 mL syringe, and the donors were allowed to rest in a holding area for 30 min. Thereafter, a second flush was performed to recover the solution remaining in the uterus. Animals from the control group were subjected to a single uterine flush. From 2 10 double-flushing procedures, 1409 viable embryos were recovered. In comparison, from 432 cows receiving the single-flushing procedure, 1993 embryos were recovered. Double flushing increased (P < 0.05) the number of embryos recovered per procedure compared to single flushing (6.7 +/- 0.4 versus 4.6 +/- 0.2, respectively; mean +/- S.E.M.). When double flushing was performed, average recovered embryos/ova increased (P < 0.05) from 8.3 +/- 0.4 to 12.7 +/- 0.7 in Limousin and from 7.9 to 11.5 in Guzera. Also, utilization of double flushing increased (P < 0.05) the number of viable embryos from 4.7 +/- 0.3 to 6.9 +/- 0.5 in Limousin and from 4.5 +/- 0.4 to 6.4 +/- 0.7 in Guzera. Mean total embryos/ova was similar (P > 0.05) between the control group and after the first uterine flushing in the double-flushing group; therefore, both flushings were conducted efficiently. In conclusion, double uterine flushing increased embryo recovery in cattle. (C) 2004 Elsevier B.V. All rights reserved.

Taxa de prenhez de vacas Nelore lactantes tratadas com progesterona associada à remoção temporária de bezerros ou aplicação de gonadotrofina coriônica eqüina

Avaliaram-se os efeitos da remoção temporária de bezerros (RTB) ou da aplicação de gonadotrofina coriônica eqüina (eCG) na taxa de prenhez (TP) de vacas Nelore lactantes tratadas com um dispositivo intravaginal liberador de progesterona (DILP). No experimento 1, 83 vacas Nelore e 102 mestiças (Nelore × Red Angus) foram distribuídas em três grupos. em dia aleatório do ciclo estral (D0), os animais do grupo 1 foram tratados com benzoato de estradiol (BE; 2,5 mg, IM, Estrogin®) e um DILP (1 g de progesterona, DIB®), removido no D9, quando também se administrou d-cloprostenol (150 µg, IM, Prolise®) e aproximadamente 12 horas após identificação do estro realizou-se a inseminação artificial. No grupo 2 (IATF), o tratamento foi semelhante ao aplicado no grupo 1, porém, administrou-se uma segunda dose de benzoato de estradiol (1 mg) no D10 e 30-36 horas mais tarde realizou-se a inseminação artificial em tempo fixo (IATF). No grupo 3 (IATF/RTB), os bezerros foram removidos a partir do D9 até a inseminação artificial com tempo fixo (IATF). As taxas de prenhez para as vacas Nelore e mestiças foram, respectivamente, de 7,69 e 41% (Controle); 23,52 e 59,57% (IATF); 69,44 e 55,81% (IATF/RTB). No experimento 2, 255 vacas Nelore lactantes foram distribuídas em três grupos. Os animais do grupo 4 (IATF) foram tratados com benzoato de estradiol (2 mg) e um DILP (1,9 g de progesterona, CIDR-B®) no D0 e no D8 sofreram remoção do DILP no D8 e administração de 25 mg de dinoprost (IM, Lutalyse®). No D9, foi aplicado benzoato de estradiol (1 mg) realizando-se a IATF 30-36 horas mais tarde. No grupo 5 (IATF/RTB), os bezerros foram removidos no D9 até a IATF. O grupo 6 (eCG) foi semelhante ao IATF, exceto pela aplicação de eCG no D9 (400 UI, IM, Novormon®). As taxas de prenhez foram de 50,57 (IATF), 53,57 (IATF/RTB) e 54,76% (eCG). A associação de RTB ao tratamento hormonal com DILP aumenta as taxas de prenhez, enquanto a adição de eCG ao tratamento não melhora as taxa de prenhez de vacas Nelore lactantes ciclando e em boa condição corporal.

LH and FSH concentration and follicular development in Nellore heifers submitted to fixed-time artificial insemination protocols with different progesterone concentrations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of temporary calf removal and eCG on pregnancy rates to timed-insemination in progesterone-treated postpartum Nellore cows

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Embryo recovery and pregnancy rates after the delay of ovulation and fixed time insemination in superstimulated beef cows

The aim of this study was to evaluate the effect of delaying ovulation subsequent to superstimulation of follicular growth in beef cows (Bos indicus) on embryo recovery rates and the capacity of embryos to establish pregnancies. Ovulation was delayed by three treatments using either progesterone (CIDR-B®) or a GnRH agonist (deslorelin). Multiparous Nelore cows (n = 24) received three of four superstimulation treatments in an incomplete block design (n = 18 per group). Cows in Groups CTRL, P48 and P60 were treated with a CIDR-B device plus estradiol benzoate (EB, 4 mg, i.m.) on Day-5, while cows in Group D60 were implanted with deslorelin on Day-7. Cows were superstimulated with FSH (Folltropin-V® 200 mg), from Day 0 to 3, using twice daily injections in decreasing amounts. All cows were treated with a luteolytic dose of prostaglandin on Day 2 (08:00 h). CIDR-B devices were removed as follows: Group CTRL, Day 2 (20:00 h); Group P48, Day 4 (08:00 h); Group P60, Day 4 (20:00 h). Cows in Group CTRL were inseminated at 10, 20 and 30 h after first detected estrus. Ovulation was induced for cows in Group P48 (Day 4, 08:00 h) and Groups P60 and D60 (Day 4, 20:00 h) by injection of LH (Lutropin®, 25 mg, i.m.), and these cows were inseminated 10 and 20 h after treatment with LH. Embryos were recovered on Days 11 or 12, graded and transferred to synchronized recipients. Pregnancies were determined by ultrasonography around Day 100. Data were analyzed by mixed procedure, Kruskal-Wallis and Chi-square tests. The number of ova/embryos, transferable embryos (mean ± S.E.M.) and pregnancy rates (%) were as follows, respectively: Group CTRL (10.8 ± 1.8, 6.1 ± 1.3, 51.5), P48 (12.6 ± 1.9, 7.1 ± 1.0, 52.3), P60 (10.5 ± 1.6, 5.7 ± 1.3, 40.0) and D60 (10.3 ± 1.7, 5.0 ± 1.2, 50.0). There were no significant differences among the groups (P > 0.05). It was concluded that fixed time AI in association with induced ovulation did not influence embryo recovery. Furthermore, pregnancy rates in embryos recovered from cows with delayed ovulation were similar to those in embryos obtained from cows treated with a conventional superstimulation protocol. © 2002 Elsevier B.V. All rights reserved.

The effect of type of vaginal insert and dose of pLH on embryo production, following fixed-time AI in a progestin-based superstimulatory protocol in Nelore cattle

The objective was to analyze and report field data focusing on the effect of type of progesterone-releasing vaginal insert and dose of pLH on embryo production, following a superstimulatory protocol involving fixed-time artificial insemination (FTAI) in Nelore cattle (Bos taurus indicus). Donor heifers and cows (n = 68; 136 superstimulations over 2 years) received an intravaginal, progesterone-releasing insert (CIDR® or DIB®, with 1.9 or 1.0 g progesterone, respectively) and 3-4 mg of estradiol benzoate (EB) i.m. at random stages of the estrous cycle. Five days later (designated Day 0), cattle were superstimulated with a total of 120-200 mg of pFSH (Folltropin-V®), given twice daily in decreasing doses from Days 0 to 3. All cattle received two luteolytic doses of PGF2α at 08:00 and 20:00 h on Day 2 and progesterone inserts were removed at 20:00 h on Day 3 (36 h after the first PGF2α injection). Ovulation was induced with pLH (Lutropin-V®, 12.5 or 25 mg, i.m.) at 08:00 h on Day 4 with FTAI 12, 24 and in several cases, 36 h later. Embryos were recovered on Days 11 or 12, graded and transferred to synchronous recipients. Overall, the mean (±S.E.M.) number of total ova/embryos (13.3 ± 0.8) and viable embryos (9.4 ± 0.6) and pregnancy rate (43.5%; 528/1213) did not differ among groups, but embryo viability rate (overall, 70.8%) was higher in donors with a DIB (72.3%) than a CIDR (68.3%, P = 0.007). In conclusion, the administration of pLH 12 h after progesterone removal in a progestin-based superstimulatory protocol facilitated fixed-time AI in Nelore donors, with embryo production, embryo viability and pregnancy rates after embryo transfer, comparable to published results where estrus detection and AI was done. Results suggested a possible alternative, which would eliminate the need for estrus detection in donors. © 2006 Elsevier Inc. All rights reserved.

Avaliação da liberação de lh e da concentração de progesterona em protocolos ovsynch e heatsynch em bubalinos (bubalus bubalis)

The objective of this study was to evaluate the LH surge after last hormonal injection of synchronization of ovulation protocols in buffalo. Fifteen multiparous buffaloes received 25 mg of Lecirelin in Day 0, and 150 mg of D-Cloprostenol on Day 7. On Day 8, estradiol benzoate was injected in Group 1 (0.5 mg, n = 5) and Group 2 (1.0 mg, n = 5). On Day 9, five buffaloes received 25 mg of Lecirelin (Control). Blood samples were collected for measure the LH concentrations on Day 7 and then every 3 hours until 72 hours after the PGF 2a injection. For evaluation of LH surge were compared the interval between PGF 2a injection to LH surge, duration, amplitude and area under the LH peak. The LH surge occurred 51.0 + 0.0 hours, 47.3 + 2.7 hours and 47.0 + 3.8 hours after PGF 2a injection for Control, Group 1 and Group 2, respectively (P > 0.05). The duration of LH peak in Control (7.8 + 1.5 hours) was shorter than Groups 1 and 2 (10.5 + 1.5 hours vs. 10.8 + 2.4 hours, respectively; P < 0.05). The amplitudes of LH peak were 4.5 + 0.4 ng/mL, 4.0 + 0.4 ng/mL and 4.3 + 0.8 ng/mL for Control, Group 1 and Group 2, respectively (P > 0.05). The area under LH peak for Control (4.8 ± 0.7) was smaller than the areas of the Groups 1 and 2 (8.8 ± 2.5 vs. 8.7 2.2, respectively; P < 0.05). In summary, the estradiol benzoate injection provided higher duration and area of LH peak than GnRH injection in Ovsynch protocol in buffalo.

LH surge in Nelore cows (Bos indicus), after induced estrus or after ovarian superestimulation

Considering that there is limited information about the preovulatory LH surge in Zebu cattle (Bos indicus), the purpose of the present work was to assess the LH surge in Nelore cows during the estrous cycle and after ovarian superestimulation of ovarian follicular development with FSH. This information is particularly important to improve superovulatory protocols associated with fixed-time artificial insemination. Nelore cows (n = 12) had their estrus synchronized with an intravaginal device containing progesterone (CIDR-B ®) associated with estradiol benzoate administration (EB, 2.5 mg, i.m., Day 0). Eight days later all animals were treated with PGF2α (Day 8) in the morning (8:00 h) and at night, when CIDR devices were removed (20:00 h). Starting 38 h after the first PGF2α injection, blood sampling and ovarian ultrasonography took place every 4 h, during 37 consecutive hours. Frequent handling may have resulted in a stress-induced suppression of LH secretion resulting in only 3 of 12 cows having ovulations at 46.7 ± 4.9 and 72.3 ± 3.8 h, respectively, after removal of CIDR-B. Thirty days later, the same animals received the described hormonal treatment associated with FSH (Folltropin ®, total dose = 200 mg) administered twice a day, during 4 consecutive days, starting on Day 5. Thirty-six hours after the first injection of PGF2α, to minimize stress, only seven blood samples were collected at 4 h interval each, and ultrasonography was performed every 12 h until ovulation. In 11 of 12 cows (92%) the LH surge and ovulation were observed 34.6 ± 1.6 and 59.5 ± 1.9 h, respectively, after removal of progesterone source. The maximum values for LH in those animals were 19.0 ± 2.6 ng/ml (mean ± S.E.M.). It is concluded that, in Nelore cows submitted to a ovarian superstimulation protocol, the LH surge occurs approximately 35 h after removal of intravaginal device containing progesterone, and approximately 12 h before the LH surge observed after an induced estrus without ovarian superstimulation. © 2008 Elsevier B.V. All rights reserved.

Sincronização do estro e da ovulação utilizando protocolos de curta duração durante a pré- estação reprodutiva em ovelhas Suffolk

This study was carried out with the objective of examining the effect of the short-term estrus synchronization protocol. Ewes were divided in four groups: Control Group (MAP sponges for 12 days, and eCG at withdrawal); Groups I, II and III used the sponge for four days, and 100 μg of PGF was applied at withdrawal; and additionally, Group I (0.1 mg of Estradiol benzoate - EB, in the sponge placement, and in the withdrawn 400 UI of eCG and 50 μg of GnRH 48h later); Group II (35 mg of injectable progesterone and 0.1 mg of EB in the sponge placement, and 400 UI of eCG at withdrawal, and 50 μg of GnRH 48h after); Group III (35 mg of injectable progesterone and 0.2 mg of EB in the sponge placement, and 400 UI of eCG at withdrawal, and 50 ?g of GnRH 56h after). Exams were accomplished for ultrasound and determine the plasmatic concentrations of progesterone and observations of the beginning the estrus and the ovulation. The lack of eCG in Group I caused this protocol to be less efficacious in induction and synchronization of estrus and ovulation. The Control Group had a greater synchronization of estrus and ovulation.

Neither plasma progesterone concentrations nor exogenous eCG affects rates of ovulation or pregnancy in fixed-time artificial insemination (FTAI) protocols for puberal Nellore heifers

The objective was to evaluate the effects of plasma progesterone (P4) concentrations and exogenous eCG on ovulation and pregnancy rates of pubertal Nellore heifers in fixed-time artificial insemination (FTAI) protocols. In Experiment 1 (Exp. 1), on Day 0 (7 d after ovulation), heifers (n = 15) were given 2 mg of estradiol benzoate (EB) im and randomly allocated to receive: an intravaginal progesterone-releasing device containing 0.558 g of P4 (group 0.5G, n = 4); an intravaginal device containing 1 g of P4 (group 1G, n = 4); 0.558 g of P4 and PGF2α (PGF; 150 μg d-cloprostenol, group 0.5G/PGF, n = 4); or 1 g of P4 and PGF (group 1G/PGF, n = 3). On Day 8, PGF was given to all heifers and intravaginal devices removed; 24 h later (Day 9), all heifers were given 1 mg EB im. In Exp. 2, pubertal Nellore heifers (n = 292) were treated as in Exp. 1, with FTAI on Day 10 (30 to 36 h after EB). In Exp. 3, pubertal heifers (n = 459) received the treatments described for groups 0.5G/PGF and 1G/PGF and were also given 300 IU of eCG im (groups 0.5G/PGF/eCG and 1G/PGF/eCG) at device removal (Day 8). In Exp. 1, plasma P4 concentrations were significantly higher in heifers that received 1.0 vs 0.588 g P4, and were significantly lower in heifers that received PGF on Day 0. In Exp. 2 and 3, there were no significant differences among groups in rates of ovulation (65-77%) or pregnancy (Exp. 2: 26-33%; Exp. 3: 39-43%). In Exp. 3, diameter of the dominant ovarian follicle on Day 9 was larger in heifers given 0.558 g vs 1.0 g P4 (10.3 ± 0.2 vs 9.3 ± 0.2 mm; P < 0.01). In conclusion, lesser amounts of P4 in the intravaginal device or PGF on Day 0 decreased plasma P4 from Days 1 to 8 and increased diameter of the dominant follicle on Day 9. However, neither of these nor 300 IU of eCG on Day 8 significantly increased rates of ovulation or pregnancy. © 2011.

Use of cooled bull semen as a strategy to increase the pregnancy rate in fixed-time artificial insemination programs-case report

Semen cryopreservation is still considered suboptimal due to lower fertility when compared to fresh semen. The reasons for the loss of fertility are various and related to irreversible damage caused to the cells during the freeze-thaw process. An alternative to conventional cryopreservation represents the use of chilled bull semen, preventing the damage associated with freezing, thereby guaranteeing greater sperm viability. The aim of this study was to describe the use of cooled bull semen as a strategy to increase the pregnancy for Fixed-Time Artificial Insemination (FTAI) of Nellore (Bos indicus) cows. One ejaculate of a select Nellore bull obtained by electroejaculation was used; the semen sample was fractioned into two aliquots: one diluted in Botu-Bov® extender containing 6.4% glycerol for cryopreservation (BB-F, frozen group) and one diluted in the same extender, free from cryoprotectants and used for cooling (BB-C, cooled semen group). The samples in the BB-C group were chilled to 5°C using an isothermic box and maintained for 24 h prior to use. A total of 349 lactating Nellore cows (70-90 days after birth) were synchronized by the insertion of a progesterone releasing device (1.0 g) and estradiol benzoate (2.0 mg i.m.) on a random day of the estrous cycle (Day 0); FTAI was performed 44-48 h after the removal of the device. The pregnancy rates were 45.71 and 61.49% (P<0.05), respectively, for the cryopreserved or chilled bovine semen groups. In conclusion, the use of bull semen cooled for 24 h represents an alternative to conventionally cryopreserved semen, as determined by the increase the pregnancy per artificial insemination in bovine herds. © 2012 Science Publication.