影响奶牛早期胚胎丢失的因素

早期胚胎丢失,即胚胎着床之前的胚胎死亡,是影响奶牛妊娠率的主要因素。奶牛妊娠早期的胚胎死亡率较高,给牧场造成了严重的经济损失。目前,对于早期胚胎丢失的研究主要集中在染色体异常、生殖激素紊乱、热应激、免疫因素、双胎妊娠、疾病、B族维生素缺乏等方面。本文综述了早期胚胎丢失发生的原因,为预防早期胚胎丢失提供依据。     

牛胚胎早期死亡的原因及预防措施

奶牛妊娠识别期(妊娠第17～18天)和妊娠黄体生成之前常发生胚胎死亡(约26%).奶牛在1次配种或授精后大约有85%以上的卵子可以受精,但是却只有55%左右的牛产犊,在整个妊娠过程中约有30%的受精卵损失,其中约25%的损失发生在受精后23天前,即胚胎死亡主要发生于配种后8～19天,另有5%的损失发生在胚胎晚期或胎儿早期.发生在奶牛中的胚胎早期死亡是使繁殖力降低和造成经济损失的主要原因.     

奶牛妊娠早期血浆中微量胎儿DNA的提取与检测

妊娠奶牛外周血的血浆中存在的胎儿游离DNA片段,为进行早期奶牛胚胎的性别诊断提供了理论依据.本试验采集妊娠奶牛静脉血液,探索从血浆中提取胎儿游离DNA的无创伤性新方法,从而为妊娠奶牛早期胚胎的性别鉴定提供新的技术途径和方法.     

奶牛胚胎死亡症的原因及综合防控

奶牛胚胎死亡症的发生是引起母牛屡配不孕的主要原因。然而长期以来,由于妊娠早期,尤其是配种后20d内的胚胎死亡后,难以准确诊断出来,致使人们对胚胎死亡症的严重性没有足够的认识。近几年来,随着超早期妊娠诊断方法的建立和不断完善,深入研究奶牛胚胎死亡症的问题显得越来越重要。因此,本文重点分析引     

影响奶牛体外性控胚胎生产效率的因素

体外受精生产性控胚胎是解决奶牛胚胎移植中胚胎来源的重要途径,卵母细胞来源和质量、分离性控精液质量、体外受精体系和早期胚胎体外培养体系是影响体外性控胚胎生产效率的主要因素。研究和探讨影响奶牛体外性控胚胎生产的重要因素,不断提高奶牛体外性控胚胎生产效率,有助于促进奶牛体外性控胚胎的商业应用。     

安胎中药方剂对奶牛胚胎着床的作用

将50头奶牛随机分为对照组、泰山磐石散组、保胎无忧散组、黄芩组及黄芩苷组5组。人工输精后7d,每天在饲料中添加不同中药散剂,连续7d。在输精后7、14、24、54d分别颈静脉采血,检测孕酮、IFN-γ和IL-4的含量。结果添加泰山磐石散、保胎无忧散的奶牛妊娠数量增加,同时显著增加血清中孕酮及IL-4水平,提高IL-4／IFN-γ比值。而补充黄芩及黄芩苷的奶牛与对照组无显著差异。推测在妊娠早期,安胎中药方剂能够调节母体内分泌及细胞因子网络,创造适宜的植入环境,利于胚胎着床。因此中药安胎方除了临床用于防治家畜流产外,还可以用于促进胚胎着床,减少早期胚胎丢失。     

奶牛早期胚胎死亡原因及分析

奶牛早期胚胎死亡是影响奶牛产奶量和繁殖率的重要因素，所以对奶牛早期胚胎死亡的原因及机理进行科学系统地分析是提高奶牛生产效率急需解决的关键问题之一。作者对引起奶牛早期胚胎死亡的内因和外因做了比较分析，从胚胎自身到母体因素及环境因素多方面进行了阐述。     

胚胎质量和发育阶段对奶牛胚胎移植妊娠率的影响

[目的]为了评估胚胎质量和发育阶段对奶牛胚胎移植妊娠率的影响。[方法]使用63头青年奶牛作为供体进行超数排卵,评估回收胚胎质量和发育阶段。选择334头青年奶牛作为受体鲜胚移植不同质量和发育阶段胚胎。对胚胎质量分布、发育阶段分布、不同质量胚胎和不同发育阶段胚胎移植30 d妊娠率进行统计分析。[结果]可用胚胎中A级胚胎比例(60.78%)显著高于B级和C级胚胎比例(36.70%和2.52%)(P<0.05);致密桑椹胚比例(54.36%)显著高于早期囊胚,囊胚和扩张囊胚比例(18.35%,25.0%和2.29%)(P<0.05)。A级和B级胚胎移植30 d妊娠率(63.55%和64.35%)显著高于C级胚胎移植30 d妊娠率(44.44%)(P<0.05);致密桑椹胚、早期囊胚、囊胚和扩张囊胚移植30 d妊娠率差异不显著(P<0.05),早期囊胚、囊胚移植30 d妊娠率高于致密桑椹胚、扩张囊胚移植30 d妊娠率(P<0.05)。[结论]选择不同发育阶段的A级和B级胚胎能够获得较高胚胎移植妊娠率,增加早期囊胚和囊胚阶段胚胎移植数量能够提高胚胎移植妊娠率。     

干扰素-τ调控牛母体妊娠识别分子机制的研究进展

奶业是农业现代化的标志性产业,在国民经济中占据重要地位。奶牛养殖是奶业发展的基础。然而,奶牛繁殖率低一直是影响奶业发展和困扰养殖户的重要问题。妊娠早期胚胎损失是导致奶牛繁殖率低的原因之一,而母体妊娠识别期是决定妊娠能否成功建立的关键环节。对妊娠识别分子机制的研究将有助于了解妊娠建立的复杂过程,可为探索减少奶牛早期胚胎损失的方法提供理论依据。本文简要综述了干扰素-τ调控牛妊娠识别分子机制的研究进展,以期为奶牛高效繁育相关科研及生产提供一定参考。     

影响胚胎移植妊娠率的因素研究

为了加快本地区农村散养奶牛品种改良的进度,本试验利用奶牛冷冻胚胎对自然发情的奶牛实施移植,研究了移植时间、解冻方法、黄体发育程度及胚胎质量等级等因素对妊娠率的影响。结果表明:胚胎移植可以像人工授精一样在奶牛自然发情状态下进行;不同移植时间、不同解冻方法、A、B级黄体及A、B级胚胎等因素对妊娠率的影响差异不显著(P>0.05)。     

Anomalous Pregnancies during Late Embryonic/Early Foetal Period in High Producing Dairy Cows

This study analyses anomalous cases of gestation ending in pregnancy loss during the early foetal period and their effect on progesterone and plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations. Data derived from a large-scale ultrasound pregnancy diagnosis programme in high producing dairy cows. Over a 3-year period (2004–2007), a very low incidence (0.5%: 15 of 3094) of anomalous pregnancies was recorded. The results revealed that the following anomalies were detected on days 35–41 of gestation in cows carrying singletons with one single corpus luteum: embryo death in eight cows (0.3%); and embryo in the uterine horn contralateral to the corpus luteum in seven cows (0.2%). All these animals suffered pregnancy loss during the early foetal period. In cows carrying dead embryos, no signs of conceptus degeneration were observed on pregnancy diagnosis. Amnion size (approximately 25 mm diameter) and uterine horn fluid contents were estimated to be similar to those of the normal pregnant cows in this period. In the contralateral gestations, live embryos were observed in all ultrasound checks before pregnancy loss. Uterine fluid contents increased in the two cows in which gestation continued for more than a week. In the cases of embryo death but not in those of contralateral gestation, a drop in PAG-1 levels was noted prior to pregnancy loss. Two cows carrying dead embryos increased with time allantoic fluid contents. The PAG-1 values increased with time in one cow bearing a dead embryo (from 2.31 to 6.79 ng/ml) and in two of the contralateral gestations (from 1.66 to 2.33 ng/ml and from 0.39 to 6.79 ng/ml, respectively). Results of this study indicate that the foetal membranes continue to undergo some activity following embryo death, and that contralateral pregnancy may determine failure of the gestation process.     

Manual Rupture Versus Transvaginal Ultrasound-guided Aspiration of Allanto-amniotic Fluid in Multiple Pregnancies: A Clinical Approach to Embryo Reduction in Dairy Cattle

To avoid the problems associated with twinning in dairy cattle, one of the embryos may be eliminated. This study compares the effect on pregnancy maintenance of two embryo reduction techniques, manual rupture (MR) and transvaginal ultrasound-guided aspiration (TUGA) of allanto-amniotic fluid, in Holstein-Friesian cows with multiple pregnancies. In the first experiment, 61 lactating cows bearing unilateral twins (n=27), bilateral twins (n=30) or triplets/quadruplets (n=4) were subjected to MR (n=45) or TUGA using a 17-G neddle (n=16) on day 28-34 of gestation. In 21 and 10 cows undergoing MR and TUGA embryo reduction, respectively, pregnancy loss occurred before day 90 (46.7 vs. 62.5%, P= 0.28). Through binary logistic regression, the type of pregnancy was identified as the only variable significantly affecting pregnancy maintenance (P=0.03). Based on the odds ratio, the risk of pregnancy loss was 4.1 times higher for unilateral twins than for bilateral twins (70.4 vs. 36.7%, respectively, P=0.01). No effect was detected on pregnancy maintenance of the technique used (P=0.17) or of the interaction technique by type of pregnancy (P=0.22). In the second experiment, a 22-G needle was used to perform TUGA on 22 lactating cows. The pregnancy loss rates were 44.4% (4/9), 18.2% (2/11) and 50% (1/2) for cows bearing unilateral twins, bilateral twins and triplets, respectively. The total pregnancy loss rate following TUGA using the 22-G needle tended to be lower than that using the 17-G needle (31.8 vs. 62.5%; P=0.06). Our results suggest that TUGA using a 22-G needle could be the method of choice to perform embryo reduction in cows carrying multiple pregnancies.     

Effect of post-insemination progesterone supplementation on pregnancy rate in dairy cows

Progesterone plays an important role in maintenance of pregnancy. It is hypothesized that insufficient progesterone early in pregnancy may result in embryonic loss, and that supplemental progesterone would decrease pregnancy loss in dairy cows. In Experiment 1, 84 cows and 16 heifers from a single dairy operation were selected randomly. Within each age category, controlled internal drug release (CIDR) devices were inserted into the vagina of every other female on Day 4 post-insemination and removed on Day 18 post-insemination. Transrectal ultrasonography was performed to determine pregnancy at 4 time periods [days 30 to 37 (week 5), days 44 to 51 (week 7), days 58 to 65 (week 9), and days 86 to 93 (week 13)]. Progesterone supplementation had no effect on pregnancy rate. In Experiment 2, there were no differences in progesterone concentrations between cows that did and did not receive a CIDR. Further, cows receiving CIDR devices did not have an increase in circulating progesterone concentrations 30 min or 1 h after CIDR insertion. It appears that progesterone supplementation does not increase circulating levels of progesterone in the early pregnant lactating dairy cow. Alterative methods to influence progesterone concentrations and/or early embryonic loss need to be investigated.     

Secondary Corpora Lutea Induced by hCG Treatment Enhanced Demi‐Embryo Survival in Lactating High‐Yielding Dairy Cows

Using a novel in vivo model considering a low developmental competence embryo (demi‐embryo) and a subnormal fertility recipient (lactating high‐yielding dairy cow), this experiment evaluated the effect of human chorionic gonadotrophin (hCG) treatment at embryo transfer (ET) on embryonic size at implantation, embryonic survival and recipient plasma progesterone (P₄) and bovine pregnancy‐specific protein B (PSPB) concentrations until day 63 of pregnancy. Embryos were bisected and each pair of demi‐embryos was bilaterally transferred to recipients (n = 61) on day 7 of the oestrous cycle. At ET recipients were randomly assigned to treatment with 1500 IU hCG or to untreated controls. Higher (p < 0.01) pregnancy rates on days 25, 42 and 63, and embryo survival rate on day 63 were observed in hCG‐treated cows with secondary CL than in hCG‐treated cows without secondary CL and in untreated cows. Pregnancy rates and embryo survival rate were similar in hCG‐treated cows without secondary CL and untreated cows. Embryonic size on day 42 was not affected by treatment with hCG, presence of secondary CL and type of pregnancy (single vs twin). Presence of secondary CL increased (p < 0.05) plasma P₄ concentrations of pregnant cows on days 14, 19 and 25 but not thereafter and of non‐pregnant cows on days 14–21. Treatment with hCG and presence of secondary CL had no effect on plasma PSPB concentrations, which were higher (p < 0.05) in twin than in single pregnancies. In conclusion, secondary CL induced by hCG treatment at ET significantly increased plasma P₄ concentrations, the survival rate of demi‐embryos and the pregnancy rate of high‐yielding lactating dairy cows. Embryos were rescued beyond maternal recognition of pregnancy, but later embryonic survival, growth until implantation and placental PSPB secretion until day 63 of pregnancy were not affected by treatment or presence of secondary CL.     

Reduced Fertility in High‐yielding Dairy Cows: Are the Oocyte and Embryo in Danger? Part I The Importance of Negative Energy Balance and Altered Corpus Luteum Function to the Reduction of Oocyte and Embryo Quality in High‐yielding Dairy Cows*

Fertility in high yielding dairy cows is declining, and there is increasing evidence to presume that oocyte and embryo quality are major factors in the complex pathogenesis of reproductive failure. In this report we present an overview of possible mechanisms linking negative energy balance (NEB) and deficiencies in oocyte and embryo developmental competence; specifically, in the high producing dairy cow. Changes in follicular growth patterns during a period of NEB can indirectly affect oocyte quality. The endocrine and biochemical changes, which are associated with NEB, are reflected in the microenvironment of the growing and maturing female gamete, and likely result in the ovulation of a developmentally incompetent oocyte. Even after an oocyte is successfully ovulated and fertilized, a full‐term pregnancy is still not guaranteed. Inadequate corpus luteum function, associated with reduced progesterone, and probably also low insulin‐like growth factor concentrations, can cause a suboptimal microenvironment in the uterus that is incapable of sustaining early embryonic life. This may partly account for the low conception rates and the high incidence of early embryonic mortality in high yielding dairy cows.     

奶牛早孕诊断技术研究进展

高繁殖率是提高奶牛场经济效益的重要保障,而早孕诊断又是提高奶牛繁殖率的重要手段之一。传统的妊娠诊断方法较难在配种后21 d内确定奶牛妊娠状态,而近年研发的新型妊娠诊断技术可以对奶牛进行有效早孕诊断。本文综述了多普勒彩超技术,中红外线光谱技术及妊娠相关蛋白、循环核苷酸、干扰素τ诱导基因检测等技术在奶牛早孕诊断中的研究进展,比较分析了这些技术的实际应用效果及优缺点,并对新的奶牛早孕诊断技术进行了展望,以期为开发和应用新型高效奶牛早孕诊断技术提供思路和借鉴。     

Pregnancy loss in dairy cattle in the Waikato region of New Zealand

AIM: To define the incidence rate of pregnancy loss and risk factors for those losses in pasture-fed dairy cattle in the Waikato region of New Zealand. METHODS: Cows (n=2,004) from 10 pasture-fed, spring-calving dairy herds in the Waikato were enrolled following confirmation of pregnancy 29-45 days after insemination, for inseminations that occurred within the first 16 days of the seasonal breeding period. Transrectal ultrasonographic examinations for pregnancy were conducted at approximately 6, 8, 10, 14 and 22 weeks gestation, and subsequent calving data were recovered. Pregnancy loss was defined as having occurred when a confirmed pregnancy was not re-diagnosed, when gross abortion was detected, or when a cow calved <265 days after the confirmed conception date. Data were analysed using reverse stepwise logistic regression and Cox's proportional hazards analysis. RESULTS: A total of 128 (6.4%) pregnancy losses were detected. The incidence rate was higher in early compared to late gestation (10.9 vs 2.8 losses/10,000 cow-days between Weeks 6-10 vs Weeks 10-14, respectively; p<0.001). Higher rates of loss were associated with the occurrence of clinical mastitis (Hazards ratio (HR)=1.57; p=0.071), being treated for anoestrus (HR=1.69; p=0.007), and in cows that had calving-to-conception intervals < or =63 days compared with those that had calving-to-conception intervals >92 days (HR=2.49; p=0.06). In addition, the rate of pregnancy loss differed between herds (p=0.05). CONCLUSIONS: The highest rate of pregnancy loss occurred in early gestation. Clinical mastitis, anoestrus and calving late in the calving season were risk factors for pregnancy loss. CLINICAL RELEVANCE: Pregnancy diagnosis using ultrasonography can be undertaken from 28 days post-insemination. However, due to the high rate of pregnancy loss at this stage of gestation, herdowners need to be warned of possible losses, and cows should be re-examined to confirm pregnancy before certification of pregnancy status is given.     

Evaluation of factors that affect embryonic loss in dairy cattle

OBJECTIVE: To identify potential risk factors for embryonic loss before 35 to 42 days of gestation in dairy cattle. DESIGN: Prospective observational study. ANIMALS: 381 cows. PROCEDURE: Body condition score was determined at the time of artificial insemination (AI; day 0) and on days 20, 23, and 27 and between days 35 and 41; serum progesterone concentration was measured on days 0; 20 or 21; and 23, 24, or 25. Cows were excluded from analyses if day 0 serum progesterone concentration was > or = 1.0 ng/mL and classified as pregnant on day 23 if serum progesterone was > 1.5 ng/mL on day 20 or 21 and day 23, 24, or 25. Cows were examined via transrectal ultrasonography on day 27 or 28 and rectally palpated for pregnancy on days 35 to 41. RESULTS: 39% of cows that were pregnant on day 23 lost their embryo by day 27, and 18% of cows that were pregnant on day 27 or 28 were not pregnant on days 35 to 41. Breeding a pregnant cow posed the greatest risk for embryonic loss at both time periods. Mean serum progesterone concentrations on day 21 or 22 and day 23, 24, or 25 were lowest for cows that lost an embryo between days 24 and 28. Cows with a linear somatic cell count score > 4.5 before AI were twice as likely to lose the embryo by 35 to 41 days, compared with cows with a score < 4.5. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that embryonic loss could be reduced by more accurate estrus detection, reducing mastitis, and strategies to improve progesterone concentration after breeding.     

Pregnancy loss in dairy cattle in the Waikato region of New Zealand

AIM: To define the incidence rate of pregnancy loss and risk factors for those losses in pasture-fed dairy cattle in the Waikato region of New Zealand.

METHODS: Cows (n=2,004) from 10 pasture-fed, spring-calving dairy herds in the Waikato were enrolled following confirmation of pregnancy 29–45 days after insemination, for inseminations that occurred within the first 16 days of the seasonal breeding period. Transrectal ultrasonographic examinations for pregnancy were conducted at approximately 6, 8, 10, 14 and 22 weeks gestation, and subsequent calving data were recovered. Pregnancy loss was defined as having occurred when a confirmed pregnancy was not rediagnosed, when gross abortion was detected, or when a cow calved <265 days after the confirmed conception date. Data were analysed using reverse stepwise logistic regression and Cox's proportional hazards analysis.

RESULTS: A total of 128 (6.4%) pregnancy losses were detected. The incidence rate was higher in early compared to late gestation (10.9 vs 2.8 losses/10,000 cow-days between Weeks 6–10 vs Weeks 10–14, respectively; p<0.001). Higher rates of loss were associated with the occurrence of clinical mastitis (Hazards ratio (HR)=1.57; p=0.071), being treated for anoestrus (HR=1.69; p=0.007), and in cows that had calving-to-conception intervals ≤63 days compared with those that had calving-to-conception intervals >92 days (HR=2.49; p=0.06). In addition, the rate of pregnancy loss differed between herds (p=0.05).

CONCLUSIONS: The highest rate of pregnancy loss occurred in early gestation. Clinical mastitis, anoestrus and calving late in the calving season were risk factors for pregnancy loss.

CLINICAL RELEVANCE: Pregnancy diagnosis using ultrasonography can be undertaken from 28 days post-insemination. However, due to the high rate of pregnancy loss at this stage of gestation, herdowners need to be warned of possible losses, and cows should be re-examined to confirm pregnancy before certification of pregnancy status is given.     

Timing of Early Foetal Loss for Single and Twin Pregnancies in Dairy Cattle

The incidence of early foetal loss is increasing under intensive management systems for dairy cattle. The aims of the present study were to determine whether there is any peak period of pregnancy loss during the early foetal period and to evaluate possible differences between single and twin pregnancies. The study population consisted of 1442 pregnant cattle from a single herd. Pregnancy was diagnosed by transrectal ultrasonography between 36 and 42 days after insemination, and then weekly until day 90 of gestation or until pregnancy loss. A total of 1310 cows (90.8%) bore single embryos and 132 (9.2%) carried twins. Pregnancy loss was registered in 139 (9.6%) cows before day 90 of pregnancy: 101 (7.7%) in single and 38 (28.8%) in twin pregnancies. The average time of pregnancy loss for all animals was 58.4 ± 12.6 days and ranged from 45 to 90 days. Seventy‐five per cent of the pregnancy losses were registered between 45 and 60 days of gestation. The average time of pregnancy loss for cows with singletons was 52.1 ± 4.1 days and ranged from 45 to 61 days and that for those with twins was 75.1 ± 12.4 days and ranged from 46 to 90 days. Seventy‐five per cent of the twin pregnancy losses were registered between 68 and 90 days of gestation. Our data show that the foetal loss in singleton pregnancies occurs earlier than in twin pregnancies. Assessment of normal development of gestation on days 60 and 90 after insemination is suggested.