Does supplemental LH changes rate and time to ovulation and embryo yield in Santa Ines ewes treated for superovulation with FSH plus eCG?

O objetivo deste trabalho foi avaliar se a suplementação com LH ao final do tratamento gonadotrófico sincroniza o tempo das ovulações e incrementa a taxa de ovulação e produção de embriões em ovelhas Santa Inês. Vinte programas de superovulação (SOV) foram realizados em delineamento cross-over (intervalo de 60 dias). No D0, um CIDR foi inserido, sendo trocado por um novo sete dias após, quando 37,5µg de d-cloprostenol foram administradas. No D12, iniciou-se o tratamento com 256mg de pFSH em 8 administrações (12/12h). No D14, o CIDR foi retirado, 200UI de eCG e 37,5µg de d-cloprostenol foram administradas. No D15, as ovelhas foram alocadas em um dos dois grupos: Controle (n=10), sem suplementação com LH, e LH (n=10), tratado com 7,5mg de LH, 24h após a remoção do CIDR. Inseminações artificiais (IA) foram realizadas 42 e 48h após a remoção do CIDR. As estruturas ovarianas foram avaliadas por laparoscopia imediatamente antes de cada IA e 5 dias após, quando os embriões foram colhidos. As ovelhas que receberam o LH tiveram maior frequência de ovulações antes de 42h (P=0,05). O tratamento com LH tendeu em incrementar a frequência de CL e diminuir a de folículos anovulatórios (P=0,08). A suplementação com LH incrementou (P=0,05) a frequência de ovelhas com alta resposta superovulatória (≥11 CL; P=0,05). em conclusão, a suplementação com LH incrementou a frequência de ovelhas com alta resposta e ovulações antes de 42h depois da remoção do CIDR, entretanto, não houve sincronia entre as ovulações. A suplementação diminuiu a frequência de folículos anovulatórios, embora a taxa de ovulação e a produção de embriões permaneceram inalteradas.

Comparison of embryo yield and pregnancy rate between in vivo and in vitro methods in the same Nelore (Bos indicus) donor cows

To investigate why the preferred means to produce bovine embryos in Brazil has changed from in vivo to in vitro, we compared these two approaches in the same Nelore cows (n = 30) and assessed total embryo production and pregnancy rates. Without a specific schedule, all cows were subjected to ultrasound-guided ovum pick up (OPU)/in vitro production (IVP) and MOET, with intervals ranging from 15 to 45 d between procedures, respectively. To produce in vivo embryos, cows were superovulated and embryos were recovered nonsurgically from 1 to 3 times (1.4 +/- 0.6). whereas OPU/IVP was repeated from 1 to 5 times (3.2 +/- 1.2) in each donor cow during a 12-mo interval. Embryos obtained from both methods were transferred to crossbred heifers. on average. 25.6 +/- 15.3 immature oocytes were collected per OPU attempt. The average number of embryos produced by OPU/IVP (9.4 +/- 5.3) was higher (P < 0.05) than the MOET method (6.7 +/- 3.7). However, pregnancy rates were lower (P < 0.05) following transfer of IVP (33.5%) versus in vivo-derived embryos (41.5%) embryos. Embryonic losses between Days 30 and 60 and fetal sex ratio were similar (P > 0.05) between in vivo and in vitro-derived embryos. We concluded that in Nelore cows, with an interval of 15 d between OPU procedures, it was possible to produce more embryos and pregnancies compared to conventional MOET. (C) 2009 Elsevier B.V. All rights reserved.

Factors affecting fertilisation and early embryo quality in single- and superovulated dairy cattle

Data on fertilisation and embryo quality in dairy cattle are presented and the main factors responsible for the low fertility of single-ovulating lactating cows and embryo yield in superovulated dairy cattle are highlighted. During the past 50 years, the fertility in high-producing lactating dairy cattle has decreased as milk production increased. Recent data show conception rates to first service to be approximately 32% in lactating cows, whereas in heifers it has remained above 50%. Fertilisation does not seem to be the principal factor responsible for the low fertility in single-ovulating cows, because it has remained above 80%. Conversely, early embryonic development is impaired in high-producing dairy cows, as observed by most embryonic losses occurring during the first week after fertilisation. However, in superovulated dairy cattle, although fertilisation failure is more pronounced, averaging approximately 45%, the percentage of fertilised embryos viable at 1 week is quite high (>70%). Among the multifactorial causes of low fertility in lactating dairy cows, high feed intake associated with low concentrations of circulating steroids may contribute substantially to reduced embryo quality. Fertilisation failure in superovulated cattle may be a consequence of inappropriate gamete transport due to hormonal imbalances.

Embryo transfer in Bos indicus cattle

In the present short review superovulation treatments commonly used for Bos taurus and/or Bos indicus will be addressed with emphasis in recent superstimulation protocols associated with pharmacological manipulation of the follicular dynamics to improve donor management and potentially embryo yield. Results obtained after superovulation treatments in which the time of LH surge is selectively delayed as an attempt to improve embryo yield are presented and discussed. (C) 2001 by Elsevier B.V.

Effect of superstimulatory treatments on the expression of genes related to ovulatory capacity, oocyte competence and embryo development in cattle.

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 AT(2) 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.

Plasma anti-mullerian hormone: an endocrine marker for in vitro embryo production from Bos taurus and Bos indicus donors

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

The effect of exogenous gonadotropins on ovarian function in goats actively immunized against inhibin

The aim of this investigation was to compare the ovarian response to superovulatory treatments in does before and after inhibin immunization, with a view to optimizing the superovulatory potential of the caprine ovary. To avoid interference by the ovarian cycle, the experiment was conducted out-of-season. At the onset of the experiment 48 does were subjected to treatment with an sc implant of the progestogen norgestomet, combined with a gonadotropin; eight does each received a single injection of 1200 IU eCG, 400 IU eCG or 2 mL physiological saline (control) or six injections (at 12 h intervals) constituting 16 or 5.4 AU pFSH. The does were mated and subjected to embryo collection 6 to 7 d later. Throughout the experiment ovarian function (by ultrasonography) and plasma levels of inhibin antibodies and progesterone were monitored. Of 40 does treated during the first part of the experiment, 48% showed estrus. The ovarian response in does treated with a high or low dose of eCG or a low dose of pFSH was barely in excess of the ovarian response in the saline-treated controls, whereas a superovulatory dose of pFSH (16 AU) gave a satisfactory response of, on average, 14.5 ovulations (yielding 8.8 flushed ova and embryos). Immediately after the does had been subjected to embryo collection they were actively immunized against inhibin by administering two injections of a recombinant α-subunit of ovine inhibin at four week intervals. All immunized does produced antibodies with the maximal titer reached two weeks after the second injection. Groups of immunized does were subjected to the same gonadotropin treatments as before (avoiding allocation of individuals to the same treatments). This time all does showed estrous symptoms. The ovulatory response to the various treatments, including the saline controls, was virtually identical, the overall average being 21.8 follicles and 9.1 ovulations. The average embryo yield per doe was 5.7. The results imply that inhibin acted as the key factor in determining the ovulatory response since no impact of any of the supplementary gonadotropins was noted in inhibin-immunized does. This finding gives rise to the notion that inhibin antibodies may act primarily by an intraovarian paracrine action rather than by reducing the suppressive action of inhibin on pituitary FSH release. Further, these findings confirm earlier reports that eCG is less suitable than FSH for inducing superovulation in goats, and indicate that active immunization against inhibin may be considered a viable alternative to using exogenous gonadotropin for inducing superovulation in goats.

Use of knowledge regarding LH receptors to improve superstimulatory treatments in cattle

Embryo transfer is a biotechnology that has been used worldwide to increase the production of offspring from female bovines. Treatments to induce multiple ovulations (superovulation) have evolved from superstimulatory protocols that depended upon detection of oestrus to treatments that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-timed AI (FTAI). The protocols associated with FTAI facilitate animal handling and produce at least as many viably embryos as conventional treatment protocols that required detection of oestrus. Recent knowledge regarding LH receptors (LHR) and follicular development can be applied to improve embryo transfer protocols. In fact, improvements in the superstimulatory treatment called the 'P-36 protocol', which include hormones that stimulate LHR, indicate that adjustments related to LHR availability may increase bovine embryo yield compared with conventional protocols based on the detection of oestrus.

Use of LH or eCG in the last day of superestimulatory treatment

In the last decades several hormonal treatments to induce multiple ovulation and embryo transfer (MOET) have been developed. Tight control of the time of ovulation allowed the use of fixed-time artificial insemination (FTAI) in embryos donors, facilitating animal management. Although, protocols that allow FTAI have evolved and yield as much embryo as conventional protocols that requires estrus detection, substantial increase in viable embryo production has not been observed in superestimulated bovine cattle. The present mini-review put emphasis on superstimulatory protocols in which the last two doses of pFSH are replaced by eCG or LH. Recent results indicate that an extra LH stimulus (using eCG or LH), on the last day of P-36 superestimulatory treatment, seems to improve transferable embryo yield in both Bos taurus and Bos indicus cattle.

Eficiência do protocolo superestimulatório P-36, associado à administração de eCG ou LH, em animais da raça Angus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Eficiência do protocolo superestimulatório P36, associado à administração de eCG ou LH, em animais da raça Nelore

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)