杜泊羊冷冻胚胎移植的影响因素

下载PDF阅读器为进一步提高杜泊羊的冻胚移植成功率,于春秋两季同期处理湖羊和小尾寒羊受体共997只(次),同期发情782只(次),发情率达78.4%,其中湖羊春秋两季平均发情率为84.6%,显著高于小尾寒羊(73.9%)(P＜0.01),秋季两品种平均发情率83.3%,显著高于春季75.8%(P＜0.01);对431只发情受体羊进行移植,总妊娠率为58.5%(252/431),产羔率为58.0%(250/431).其中湖羊为受体的春秋两季平均妊娠率62.5%,高于小尾寒羊(54.7%);秋季两品种的平均妊娠率为64.8%,显著高于春季(52.5%).囊胚移植后的平均妊娠率为61.5%,显著高于桑椹胚(41.5%),其中A级囊胚为67.4%,显著高于B级囊胚(P＜0.05)和所有桑椹胚(P＜0.01);B级囊胚显著高于B级桑椹胚(P＜0.05),B级囊胚与A级桑椹胚间及A、B级桑椹胚间均无显著差异;C级囊胚和桑椹胚均未见妊娠.此外,受体羊卵巢黄体发育状况及移植技术熟练程度对妊娠率也有显著影响.

受体品种、体重及移植季节对杜泊羊胚胎妊娠率和新生羔羊体重的影响

胚胎移植可以提高家畜良种扩大的速度, 但移植成功率会受到很多因素的影响。用615～715 日龄 杜泊羊冷冻胚胎作为移植胚胎, 研究了季节和受体状况对杜泊羊胚胎移植妊娠率以及出生羔羊生长发育的影 响。在比较春秋两季236 枚冷冻胚胎的移植效果发现: 秋季移植的妊娠率(6816 %) 显著高于春季(5815 %) ; 春季和秋季移植后出生羔羊的体重, 初生时无显著差异; 但在30 日和60 日(断乳) 时, 春季移植的羔羊显著 高于秋季。比较不同品种的受体发现, 昭通绵羊受体的胚胎移植妊娠率最高, 湖羊次之, 小尾寒羊最低, 但受 体羊3 种品种间无显著差异; 较大受体羊( ≥40 kg) 生产的羔羊出生和30 d 的体重显著高于较小受体( < 40 kg) ; 但60 d 羔羊体重差异不显著, 说明受体体重会影响羔羊的出生体重, 但是对哺乳期的生长发育影响不大。 春季移植出生的羔羊, 其早期生长发育情况明显优于秋季。

影响杜泊羊胚胎移植妊娠率和新生个体生长发育的因素：受体和移植季节

胚胎移植具有加速家畜良种推广的潜力。胚胎移植能直接引入纯种动物，并且能避免直接进口动物时可能会带来一些较大传染病（例如口蹄疫）的风险，因此具有成本低、安全性高、直接引入良种的特点。但是胚胎移植的成功率可能会受到很多因素的影响。这些因素可归纳为三个方面：胚胎因素；胚胎和受体的相互作用；受体因素。另一方面，胚胎移植目的是为了获得优秀生产性能的良种个体，但是出生的后代的生长发育情况也会和很多因素有关，例如出生大小、当地气候条件、饲养条件和胚胎移植季节等。因此，研究影响胚胎移植妊娠率以及出生后代生长发育的因素对胚胎移植在生产中的应用有重要意义。杜泊羊作为一种肉用绵羊，品种优良，经济价值很高，被认为应予以积极推广。用胚胎工程对杜泊羊进行推广可能是一个较好的办法。绵羊胚胎工程近年来已经取得了较大进展，例如超数排卵、体外胚胎生产、胚胎冷冻和胚胎移植的研究都取得了一系列的成果。在影响绵羊胚胎移植效果的具体因素中，已有一些关于不同胚胎移植数目、胚龄、冻胚和鲜胚、胚胎移植方法等报道，但是受体品种对杜泊羊的胚胎移植的研究还未见报道，季节对杜泊羊胚胎移植的影响的研究结果也不一致。因此本实验应用质量稳定的胚胎，采用单胚移植，以移植的妊娠率为指标，研究了受体品种和季节因素对杜泊羊冷冻胚胎移植成功率影响，结果发现在三个不同品种的受体中，胚胎移植妊娠率无显著差异（P＞0．05）；但是季节对胚胎移植成功率有显著影响，秋季移植妊娠率（68.6％）显著高于春季（58.5％，P＜0.05）。在山羊等其他动物的研究中表明，受体情况不影响胚胎移植出生的后代的生长发育。本实验以羔羊的出生重、1个月体重和断乳时（2个月时）体重为指标，分析了受体体重和季节对胚胎移植出生杜泊羊羔羊的早期生长发育的影响，结果表明，不同体重的受体会影响羔羊的出生重（P＜0.05），即体重较大的受体出生的羔羊的体重也较大，但经过两个月的哺乳期生长后，羔羊的生长发育情况已无明显差异（P＞0.05）。有研究表明羔羊早期的生长发育情况和外界条件有关。不同季节进行胚胎移植，羔羊出生时间也不同。因此，实验研究了不同季节对胚胎移植出生羔羊早期生长发育的影响。结果发现，季节显著影响了羔羊的生长发育，春季和秋季移植出生的羔羊在出生重方面差异不显著（P＞0.05），但春季移植出生的羔羊的后来的生长情况要明显好于秋季（P＜0.05）。

Impact of various endocrine and paracrine factors on in vitro culture of preantral follicles in rodents

Folliculogenesis is a complex process regulated by various paracrine and autocrine factors. In vitro growth systems of primordial and preantral follicles have been developed for future use of immature oocytes, as sources of fertilizable oocytes and for studying follicular growth and oocyte maturation mechanisms. Rodents were often chosen for in vitro follicular culture research and a lot of factors implicated in folliculogenesis have been identified using this model. To date, the mouse is the only species in which the whole process of follicular growth, oocyte maturation, fertilization and embryo transfer into recipient females was successfully performed. However, the efficiency of in vitro culture systems must still be considerably improved. Within the follicle, numerous events affect cell proliferation and the acquisition of oocyte developmental competency in vitro, including interactions between the follicular cells and the oocyte, and the composition of the culture medium. Effects of the acting factors depend on the stage of follicle development, the culture system used and the species. This paper reviews the action of endocrine, paracrine factors and other components of culture medium on in vitro growth of preantral follicles in rodents.

Morpho-physical Recording of Bovine Conceptus (Bos indicus) and Placenta from Days 20 to 70 of Pregnancy

The study is based on 141 pregnant Bos indicus cows, from days 20 to 70 post-insemination. First, special attention was given to the macroscopically observable phenomena of attachment of the conceptus to the uterus, i.e. the implantation, from about days 20 to 30 post-insemination up to day 70, and placentome development by growth, vascularization and increase in the number of cotyledons opposite to the endometrial caruncles. Secondly, as for the conceptuses, semiquantitative, statistical analyses were performed of the lengths of chorio-allantois, amnion and yolk sac; and the different parts of the centre and two extremes of the yolk sacs were also analysed. Thirdly, the embryos/foetuses corresponding to their membranes were measured by their greatest length and by weight, and described by the appearance of external developmental phenomena during the investigated period like neurulation, somites, branchial arcs, brain vesicles, limb buds, C-form, pigmented eye and facial grooves. In conclusion, all the data collected in this study from days 20 to 70 of bovine pregnancy were compared extensively with corresponding data of the literature. This resulted in an `embryo/foetal age-scale`, which has extended the data in the literature by covering the first 8 to 70 days of pregnancy. This age-scale of early bovine intrauterine development provides model for studies, even when using slaughtered cows without distinct knowledge of insemination or fertilization time, through macroscopic techniques. This distinctly facilitates research into the cow, which is now being widely used as `an experimental animal` for testing new techniques of reproduction like in vitro fertilization, embryo transfer and cloning.

Simulação e análise econômica da produção in vivo e in vitro de embriões em bovinos

The objective of this work was to evaluate the effect of the variables number of recipients, synchronization protocol, reproductive efficiency indicators and pregnancy cost, in the economic effectiveness of in vivo and in vitro bovine embryo production. A simulation application was elaborated to allow the user to insert the input variable parameters. A basic scenario, from the efficiency traditional rates of in vivo (ET) and in vitro production (IVP) techniques of bovine embryos, was introduced in the software as a criterion to compare the results. This software was able to reproduce both ET and IVP scenarios. The embryo production was simulated through stochastic simulation. The optimal number of recipients using sensitivity analysis was determined. The net present value and cost per pregnancy were used as a decision parameter. The synchronization for fixed-time embryo transfer decreased the recipient idleness and, consequently, the final cost of pregnancy, in comparison to the traditional methodology. Foetal sexing must be associated to IVP of bovine embryos. In addition, the optimal recipient number per donor is variable and depends on data inserted in the system.

Correlação entre os métodos de concepção, ocorrência e formas de tratamento das onfalopatias em bovinos: estudo retrospectivo

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

Zona thinning with a noncontact diode laser in ICSI embryos from women of advanced age

Purpose: the objective of this study was to determine if the zona thinning (ZT) technique improved the rates of implantation and clinical pregnancy for patients aged, greater than or equal to38 years submitted to an ICSI program.Methods: A total of 100 patients submitted to ICSI and aged, greater than or equal to38 years were divided in a prospective and randomized manner into two groups: Group I - patients submitted to ZT (n = 50); a laser diode with 1.48 mum wavelength (Fertilaser) was used for the ZT procedure with 1-2 irradiations of 10 ms applied to four different positions on the zona pellucida (ZP) of each embryo to thin 60-90% of the ZP (each point with a 15-20 mum length of ZT). Group II - patients with no ZT (n = 50). In both groups, embryo transfer was performed on the second or third day.Results: the age of Group I patients (39.8 +/- 1.3) did not differ (p = 0.67) from that of Group II patients (40 +/- 1.9). The number of oocytes retrieved at metaphase II from Group I (6.4 +/- 4.2) and Group II (6.8 +/- 5) was similar (p = 0.94). Normal fertilization rates and cleavage rates were similar (p = 0.78 and p = 0.63, respectively) for Group I (71.5 +/- 22% and 96.7 +/- 11%) and Group II (73.5 +/- 19.7% and 96 +/- 11%, respectively). The number of embryos transferred was similar (p = 0.53) for the two groups (Group I = 3.1 +/- 1.3; Group II = 2.9 +/- 1.1). The thickness of the ZP of Group I embryos (16.9 +/- 2.4 mum) did not differ (p = 0.97) from that of Group II embryos (16.9 +/- 2.3 mum). The rates of embryo implantation and clinical pregnancy per embryo transfer were similar (p = 0.67, p = 0.61) for Group I (7 and 16%, respectively) and for Group II (8.2 and 22%, respectively).Conclusions: These results suggest that ZT in the population aged, 38 years may have no impact on ICSI success rates. However, this conclusion is limited to a situation in which length of the laser ZT was less than or equal to 20 mum and the laser was applied to four different positions.

Are poor responders patients at higher risk for producing aneuploid embryos in vitro?

Purpose To test the hypothesis that aged women with poor ovarian response express an increase on embryo chromosomal alterations when compared to aged women who presented normal response.Methods Couples undergoing intracytoplasmic sperm injection cycles with preimplantation genetic screening, were subdivided into two groups: Poor Responder group (n = 34), patients who produced a parts per thousand currency sign4 oocytes; and Normoresponder group (n = 50), patients who produced a parts per thousand yen5 oocytes. Groups were compared regarding cycles' outcomes and aneuploidy frequency.Results There were no significant differences between and groups regarding the fertilization rate (p = 0.6861), clinical pregnancy (p = 0.9208), implantation (p = 0.6863), miscarriage (p = 0.6788) and the percentage of aneuploid embryos (p = 0.270). Embryo transfer rate was significantly lower on poor responder group (p = 0.0128) and logistic regression confirmed the influence of poor response on the chance of embryo transfer (p = 0.016).Conclusions Aged females responding poorly to gonadotrophins are not at a higher risk for producing aneuploid embryos in vitro.

Factors potentially affecting fertility of lactating dairy cow recipients

Objectives of this study were to evaluate factors that could affect pregnancy rate after embryo transfer (ET) in lactating dairy cow recipients. The trial was conducted at a dairy farm located in Descalvado, SP, Brazil from October 2003 to September 2004. From 1037 cows with CL that were treated with an injection of PGF(2 alpha), 43.3% were detected in heat; 263 were previously assigned at day of PGF(2 alpha) injection for AI and 186 for ET. Ovulation rate was 85.7% (385/449). Pregnancy rate for cows with CL for AI and embryo transfer recipients were 36.5% (84/230) and 58.7% (91/155) at day 25 and 33.0% (76/230) and 45.8% (71/155) at day 46, respectively. Embryonic loss were 9.5% (8/84) for the AI group and 21.9% (20/91) for the ET group. Average milk production was 31.4 L/day/cow. Average daily milk production from 7 days before PGFZ injection to 7 days after ET tended (P < 0.10) to influence pregnancy rate on days 25 and 46. Average daily milk production from the day of embryo transfer to 7 days after influenced embryonic loss (P < 0.05). Cows with higher milk production had lower probability of pregnancy and higher probability of embryonic loss. Cows with higher days in milk had higher probability of pregnancy. Cows with higher rectal body temperature had lower probability of pregnancy and higher probability of embryonic loss. The influence of high milk yield and body temperature on fertility in lactating dairy cow recipients suggests that these effects can occur also after embryo reaches the blastocyst stage. (c) 2005 Elsevier B.V. All rights reserved.

Associations among milk production and rectal temperature on pregnancy maintenance in lactating recipient dairy cows

The objective of this study was to evaluate the associations among milk production, rectal temperature, and pregnancy maintenance in lactating recipient dairy cows. Data were collected during an 11-mo period from 463 Holstein cows (203 primiparous and 260 multiparous) assigned to a fixed-time embryo transfer (ET) protocol. Only cows detected with a visible corpus luteum immediately prior to ET were used. Rectal temperatures were collected from all cows on the same day of ET. Milk production at ET was calculated by averaging individual daily milk production during the 7 d preceding ET. Pregnancy diagnosis was performed by transrectal ultrasonography 21 d after ET. Cows were ranked and assigned to groups according to median milk production (median = 35 kg/d; HPROD = above median; LPROD = below median) and rectal temperature (<= 39.0 degrees C = LTEMP; >39.0 degrees C = HTEMP). A milk production x temperature group interaction was detected (P = 0.04) for pregnancy analysis because HTEMP cows ranked as LPROD were 3.1 time more likely to maintain pregnancy compared with HTEMP cows ranked as HPROD (P = 0.03). Milk production did not affect (P = 0.55) odds of pregnancy maintenance within LTEMP cows, however, and no differences in odds of pregnancy maintenance were detected between HTEMP and LTEMP within milk production groups (P > 0.11). Within HTEMP cows, increased milk production decreased the probability of pregnancy maintenance linearly, whereas within LTEMP cows, increased milk production increased the probability of pregnancy maintenance linearly. Within HPROD, increased rectal temperature decreased the probability of pregnancy maintenance linearly, whereas within LPROD cows, no associations between rectal temperatures and probability of cows to maintain pregnancy were detected. In summary, high-producing dairy cows with rectal temperatures below 39.0 degrees C did not experience reduced pregnancy maintenance to ET compared to cohorts with reduced milk production. (C) 2011 Elsevier B.V. All rights reserved.

Effects of rumen-protected polyunsaturated fatty acid supplementation on reproductive performance of Bos indicus beef cows

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

Effects of calcium salts of polyunsaturated fatty acids on productive and reproductive parameters of lactating Holstein cows

Two experiments evaluated milk production, serum progesterone and insulin, and reproductive performance of lactating Holstein cows receiving or not receiving Ca salts of polyunsaturated fatty acids (PUFA), or receiving Ca salts of PUFA at different daily frequencies. In experiment 1, 1,125 cows randomly distributed in 10 freestall barns were enrolled. Barns were assigned randomly to receive a high-concentrate diet containing (PI?) or not containing (control, CON) 1.1% (dry matter basis) Ca salts of PUPA. Diets were offered 6 times daily, whereas the Ca salts of PUFA were included in the PF treatment in the first feeding of the day. In experiment 2, 1,572 cows were randomly distributed in 10 freestall barns, which were assigned randomly to receive a diet similar to PF, but with Ca salts of PUFA included only in the first feeding of the day (PF1X), or equally distributed across all 6 feedings (PF6X). During both experiments, cows were artificially inseminated 12 h after the onset of estrus. Once per month, cows that did not conceive to artificial insemination were assigned to a fixed-time embryo transfer protocol. Pregnancy was determined via transrectal ultrasonography 28 and 60 d after expected ovulation. Pregnancy loss was considered in cows that were pregnant on d 28 but nonpregnant on d 60. During both experiments, feed intake, milk yield, and milk protein and fat content were recorded weekly. Blood samples were collected concurrently with embryo transfer. During experiment 1, feed intake was similar between treatments. Compared with CON, PF cows had greater milk yield (37.8 vs. 35.3 kg/d), and reduced milk fat content (3.41 vs. 3.55%). However, PF cows had reduced pregnancy losses per service compared with CON (12.6 vs. 18.3%). Serum progesterone was greater and serum insulin tended to be greater in primiparous cows receiving PF compared with CON cohorts (4.50 vs. 3.67 ng of progesterone/mL, and 10.4 vs. 7.5 mu UI of insulin/mL). During experiment 2, no treatment effects were detected for feed intake, milk yield, or milk fat, whereas PF1X cows tended to have reduced pregnancy losses per service compared with PF6X (14.4 vs. 18.4%). In summary, feeding Ca salts of PUFA to dairy cows increased milk production, did not alter feed intake, and reduced pregnancy losses per service. Further, the total daily amount of Ca salts of RITA should be fed during the first feeding of the day to optimize its benefits on pregnancy maintenance of dairy cows.