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1.
本研究的目的是探讨水牛分离精子与不同来源(活体采卵或屠宰场卵巢采卵)卵母细胞体外受精的效果。活体采卵是选用20头空怀河流型母水牛(其中摩拉母牛12头,尼里-拉菲母水牛8头)每间隔3 d采卵1次,连续采卵5~6周,活体采集卵母细胞;屠宰场卵巢采卵是收集屠宰场水牛卵巢,用10 mL注射器连接18 G针头吸取水牛卵巢上可视的卵泡来收集卵母细胞。将收集的AB级水牛卵母细胞在相同的条件下进行体外成熟、然后用分离或未分离精子进行体外受精以及体外培养至囊胚。结果发现:活体采卵组和屠宰场收集的水牛卵母细胞组用分离精子受精分裂率和囊胚率没有差异(P>0.05);分离精子和未分离精子的体外受精分裂率和囊胚率也没有差异(P>0.05)。由此说明,水牛分离精子可以用于体外生产性控胚胎。  相似文献   

2.
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3.
Oocytes from abattoir-sourced ovine ovaries were aspirated from 2- to 4-mm follicles using 25, 50 or 100 mmHg pressure and an aspiration pump, or a needle (20-G) and syringe (2.5 ml) and subjected to in vitro maturation, fertilization and culture to determine the effect of aspiration pressure on the number and quality of oocytes recovered, and early embryonic development. Oocyte recovery rate was similar between groups (range: 57.1-73.1%; p > 0.05). The number and percentage of grade I and II oocytes recovered was reduced for 100 mm (24.5 +/- 3.6 and 31.1 +/- 6.1%) compared with 25 mm (51.4 +/- 7.0 and 60.2 +/- 7.8%) and 50 mm pressure (40.8 +/- 5.6 and 50.3 +/- 4.4%) and a syringe (40.3 +/- 12.0 and 45.2 +/- 2.1%; p < 0.05). Oocyte cleavage was similar for all groups at 24 (range: 30.9-49.6%) and 48 h post-insemination (49.7-65.5%), but blastocyst formation (% cleaved oocytes) was lower for oocytes aspirated with 25 mm (37.8%) than 50 (69.2%) or 100 mm (67.2%) pressure, and a syringe (72.0%; p < 0.05). Embryo production efficiency (% of oocytes cultured developing to the blastocyst stage) was higher for oocytes aspirated with 50 mm (45.4%) and 100 mm pressure (43.8%) and a syringe (45.0%) than 25 mm pressure (18.8%; p < 0.05). These results demonstrate that the aspiration of ovine oocytes with an aspiration pressure of 50 mm, or aspiration with a needle and syringe are equally efficacious for the in vitro production of embryos.  相似文献   

4.
This study was designed with the final goal of improving in vitro embryo production in the Thai swamp buffalo (Bubalus bubalis carabensis). Oocytes were collected by ovum pick-up (OPU) from six non-lactating multiparous swamp buffalo twice per week for 10 consecutive sessions followed by once-weekly collection for 10 consecutive sessions without hormone stimulation. In addition, oocytes were collected from slaughterhouse ovaries that were classified as follows: ovaries from non-pregnant cows with a visible corpus luteum (NPCL); pregnant cows with a corpus luteum (P); and non-pregnant cows without a corpus luteum (NP). Follicles in each group of ovaries were categorized as small (2-4 mm), medium-sized (5-8 mm) or large follicles (≥ 9 mm). The quality of the oocytes was assessed by their capacity to undergo in vitro maturation. The total number of observed follicles per session (all sizes combined) was larger in the once-weekly OPU group compared with the twice-weekly OPU group. In particular, the numbers of small and large follicles were higher in the once-weekly OPU group (5.2 ± 0.7 and 0.9 ± 0.2, respectively) than in the twice-weekly OPU group (3.9 ± 0.5 and 0.5 ± 0.1). The number of medium-sized follicles did not differ between the groups. The percentages of oocytes with an abnormal spindle morphology were not different between oocytes from the twice-weekly (30.0%) and the once-weekly (28.6%) OPU groups. A higher percentage of oocytes obtained in vitro (49.5%) exhibited nuclear abnormalities compared with those obtained in vivo (≤34.8%) after in vitro maturation. In conclusion, oocytes can be successfully collected by OPU in the swamp buffalo, without hormonal pretreatment, and per week more good-quality oocytes can be collected by twice-weekly OPU. In addition, oocytes collected from slaughterhouse ovaries can be used with the reproductive status of the cow having no influence on the maturation competence of oocytes.  相似文献   

5.
The objective of this study was to evaluate the efficiency of Ovum Pick Up (OPU) in cycling (n = 5) and lactating, postpartum, swamp buffaloes (n = 6) with and without gonadotropin stimulation. The OPU was performed every two weeks in all groups of animals, for a total of six sessions. Thirty collections were performed in five cycling buffaloes and 36 collections in six lactating postpartum buffaloes. Buffaloes that received hormonal stimulation were given a total of 400 mg, follicle stimulating hormone (FSH), administered twice daily over 3 days in decreasing doses, together with 100 microg of GnRH, 24 h after the last FSH injection. Following a resting period of 1 month, the two groups of buffaloes, were subjected to the same OPU regimen, but without any hormonal treatment for an additional six OPU sessions. The number of aspirated follicles recorded from the hormonal stimulated, cycling animals and lactating, postpartum buffaloes was not significantly different, 7.2 +/- 3.7 and 9.0 +/- 3.2, respectively (p > 0.05). Recovered oocytes collected from the two groups of hormonally stimulated animals were also not statistically different: 3.7 +/- 2.7 in the cycling and 5.9 +/- 3.5 in the lactating postpartum group (p > 0.05). In the two groups of buffaloes not receiving hormonal stimulation, the number of aspirated follicles was not significantly different: 2.1 +/- 1.4 and 1.4 +/- 0.7 in cycling and lactating postpartum buffaloes respectively (p > 0.05). Recovered oocytes in the non-treated groups were also similar: 1.4 +/- 1.3 vs 0.7 +/- 0.8 in cycling and lactating buffaloes (p > 0.05). Among stimulated buffaloes, most aspirated follicles were small in size (< or =5 mm), whereas they were mostly medium and large sizes in the non-treated buffaloes. The oocyte recovery rate in both the groups, cycling and lactating postpartum, were 51.6% and 69.5% in stimulated groups and 55.0% and 53.1% in non-stimulated groups (p > 0.05). The majority of recovered oocytes were single- and multi-layered, and the number was greater in the cycling than in the lactating, postpartum buffaloes. The number and quality of recovered oocytes was similar in all groups of buffaloes whether they were received or did not receive hormonal stimulation. Moreover no difference was found in multi- and single-layered oocytes between cycling and lactating, postpartum buffaloes. In conclusion, OPU can be performed successfully in swamp buffalo in different reproductive status and FSH administration was shown to increase the number of aspirated oocytes in both cycling and lactating, postpartum buffaloes.  相似文献   

6.
In this study, we examined the effects of superstimulation using follicle‐stimulating hormone (FSH) followed by gonadotropin‐releasing hormone (GnRH) on buffalo embryo production by ultrasound‐guided ovum pick‐up (OPU) and in vitro fertilization (IVF). Nine Murrah buffaloes were subjected to OPU‐IVF without superstimulation (control). The morphologies of the oocytes collected were evaluated, and oocytes were then submitted to in vitro maturation (IVM). Two days after OPU, same nine buffaloes were treated with twice‐daily injections of FSH for 3 days for superstimulation followed by a GnRH injection. Oocytes were collected by OPU 23–24 hr after the GnRH injection and submitted to IVM (the superstimulated group). The total number of follicles, number of follicles with a diameter > 8 mm, and number of oocytes surrounded by multi‐layered cumulus cells were higher in the superstimulated group than in the control group (p ≤ 0.05). After IVF, the percentages of cleavage and development to blastocysts were higher in the superstimulated group than in the control group (p < 0.05). In conclusion, superstimulation improved the quality of oocytes and the embryo productivity of OPU‐IVF in river buffaloes.  相似文献   

7.
This work was designed to evaluate the ovarian follicular development, oocytes morphology, methods of oocytes reterival, and the effect of different in vitro maturation (IVM) media on cumulus cell expansion and nuclear maturation of Jennies oocytes. Experiment 1, the number of small (<6 mm), medium (6 to 9 mm) and large size (>10 mm) ovarian follicles was recorded. Cumulus-oocyte-complexes (COCs) were reterived and classified into 4 Grades based on their cumulus-cells investment and the homogenous of the ooplasm. In Experiment 2, COCs were recovered by using 18-G, 20-G needle or slicing and scraping of ovarian follicles to determine the number and morphology of the recovered COCs. In Experiment 3, Grade A and B COCs were IVM in DMEM-HG, DMEM-LG, DMEM-F12, TCM199, TCM199-F12 or CR1aa media supplemented with 10 % FCS?+?10 μg FSH/mL?+?10 IU hCG/mL?+?50 μg/mL gentamicin. Maturation was performed for 36 h at 38.5 °C under 5 % CO2 in humidified air. After IVM, cumulus cell expansion and oocytes nuclear canfiguration were determined. An average of 6.40?±?0.26 follicles was recorded per Jenny ovary, representing 3.37?±?0.46, 1.89?±?0.14 and 1.14?±?0.16, for the small, medium and large size follicles, respectively. Oocyte recovery was higher (P?P?P?P?P?P?P?P?Conclusion: Slicing and scraping or aspiration of follicles using 18-G needle increased the number and percentage of Grade A Jennies oocytes. TCM199-F12, CR1aa and TCM199 medi are more suitable for IVM of Jenny oocytes by promoting cumulus cells expansion and nuclear maturation to M II stage.  相似文献   

8.
Ovum pick-up (OPU) was performed three times on adult ewes after synchronization with (n = 4) or without (n = 4) FSH treatment to investigate the effects of FSH treatment on the number of ovarian follicles, oocytes recovered, oocyte quality and development in vitro. FSH treatment increased the number of ovarian follicles (85 vs 162) and oocytes recovered (33 vs 91), although recovery rate was similar for ewes with and without FSH (91/162, 56.2% and 33/85, 38.8% respectively). Of the oocytes recovered, those classified as grades I and II were similar between ewes with (78/91, 85.7%) and without FSH treatment (27/33, 81.8%). The number of ovarian follicles was similar after repeated OPU for ewes treated with FSH, but for ewes not treated with FSH the number of ovarian follicles decreased with repeated OPU. The number of oocytes recovered decreased for FSH-treated ewes only, while the oocyte recovery rate and proportion of oocytes classified as grades I and II were not affected by repeated OPU. Oocyte cleavage (46/78, 58.9% and 19/24, 79.2%) and blastocyst formation (35/46, 76.1% and 12/19, 63.2% respectively) were similar for ewes with and without FSH treatment. The number of ovarian follicles varied between ewes (p < 0.05) although the number of oocytes recovered and oocyte development in vitro were similar between ewes.  相似文献   

9.
高产奶牛连续活体采卵及卵母细胞体外受精   总被引：8，自引：1，他引：7  
在超声波扫描仪的指导下,用双孔型采卵针以 １４．７ ｋ Ｐａ 抽吸压经阴道对 ５ 头高产奶牛分别连续实施 ７ 次活体采卵,每 ７ ｄ １ 次,共采集卵子 ２１４ 个,占可见卵泡数的 ５３．９％ ,每次头均采集卵子 ６．１ 个。卵子经过体外培养、体外受精、体外受精胚体外发育培养,于体外受精后 ４８ ｈ 、１６８ ｈ 统计的卵裂率、囊胚发育率分别为 ７５．２％ 和 ２９．７％ 。研究结果表明,在超声波扫描仪指导下对奶牛连续进行活体采卵是可行的,所得卵子应用体外成熟、体外受精、体外培养技术,可生产用于冷冻或移植的胚胎。  相似文献   

10.
Embryo production in donkeys is inefficient compared with that in other livestock. Obtaining a sufficient number of MII oocytes is the first step to solving this problem. In this study, the number, morphology and maturation rates of cumulus-oocyte complexes (COCs) obtained from abattoir-derived ovaries or live jennies were compared. The diameter of follicles from abattoir-derived ovaries was measured and divided into group 1 (2–6 mm), group 2 (6–10 mm), group 3 (10–20 mm), group 4 (20–28 mm) and group 5 (>28 mm). The results showed that the number of follicles per ovary in group 2 (3.6 ± 0.28) and 3 (4.2 ± 0.90) was higher than that in the other groups (p < .05). The recovery rate in group 3 was higher than group 1 (48.8% vs. 26.8%, p = .00), but lower than group 5 (48.8% vs. 76.5%, p = .025). The percentage of grade A COCs in group 3 was higher than group 2 (59.3% vs. 39.5%, p = .00) and group 1 (59.3% vs. 26.7%, p = .00). Moreover, the percentage of grade A COCs in group 4 (55.0%, p = .710) and group 5 (46.2%, p = .351) was reduced compared with that in group 3. From the above results, the developing follicles (group ovum pick-up [OPU], 10–20 mm) and preovulation follicles (group OPU-Preov, >35 mm) were aspirated from live jennies using OPU. Although there was no difference in the recovery rates of COCs between group 3 and OPU (48.8% vs. 43.0%, p = .184), the percentage of grades A COCs in group OPU was higher than group 3 (72.5% vs. 59.3%, p = .036). There was no difference in the maturation rate between group 3 and OPU (60.3% vs. 69.3%, p = .171) after the COCs matured in vitro. The rates of recovery (72.2%) and maturation (92.3%) in group OPU-Preov were higher than those in other groups (p < .05). Moreover, the effects of maturation time and serum type on maturation rates were evaluated in groups B44 (44 h, FBS), B36 (36 h, FBS) and D44 (44 h, foetal donkey serum, FDS). These results indicated that the maturation rate in group B36 was lower than group B44 (13.1% vs. 47.0%, p = .00) and group D44 (13.1% vs. 53.3%, p = .00). In conclusion, the quality of donkey COCs from OPU was higher than that from abattoir-derived ovaries, the suitable time of donkey in vitro maturation (IVM) was 44 h, and FBS could be replaced with FDS in donkey IVM medium.  相似文献   

11.
The effects of twisting and type (single- or double-lumen) of aspiration needle on the efficiency of transvaginal ultrasound-guided ovum pick-up (US-guided OPU) were investigated in cattle. The first study using slaughterhouse ovaries revealed that twisting of the needle during follicle aspiration improved the oocyte recovery rate without deleterious effects on the attachment of cumulus layers. Vacuum pressure affected the oocyte recovery and cumulus attachment, regardless of the needle type. The needle type did not affect the oocyte recovery or cumulus attachment with an optimized vacuum pressure. In the second study, US-guided OPU was performed in live cows using two types of needles with a vacuum pressure of 75 mmHg. The needle type did not affect the oocyte recovery or cumulus attachment of the recovered oocytes. The results revealed that twisting of the needle is effective in follicle aspiration, and suggested that a single-lumen needle is as useful as a double-lumen needle for US-guided OPU in cattle.  相似文献   

12.
【目的】探索不同来源卵母细胞对体细胞核移植(SCNT)重构胚的发育能力及发育潜能关键蛋白表达水平的影响。【方法】试验分为活体采卵(OPU)和屠宰场(SLH)卵巢2组,OPU组用超声波活体采卵仪穿刺抽吸10头非泌乳期经产水牛卵巢的卵泡采卵,SLH组从屠宰场卵巢抽吸卵泡采卵。获得的卵母细胞分别进行体外成熟,体外成熟22～24 h后,吹打去除卵丘细胞,挑选具有第一极体的卵母细胞,去核后与水牛耳部成纤维细胞进行SCNT,分别统计SCNT重构胚的融合率、分裂率和囊胚率,用免疫荧光检测2种SCNT重构胚的E-钙黏蛋白(E-cadherin)和转录因子Sox2蛋白的表达水平。【结果】OPU组卵母细胞成熟率及其SCNT重构胚的囊胚率均显著高于SLH组(P<0.05),但2组SCNT重构胚的融合率和分裂率均无显著差异(P>0.05);免疫荧光结果显示,E-cadherin蛋白定位于细胞膜上,Sox2蛋白分布在细胞核膜及细胞质中,OPU组SCNT重构胚中E-cadherin和Sox2的表达水平均显著高于SLH组(P<0.05)。【结论】活体采集的水牛卵母细胞更适合用于SCNT重构胚的构建...  相似文献   

13.
There are indications in the literature that delaying the period between ovarian superestimulation and ovum pick up (OPU) would induce follicles to a condition of initial atresia, which could be beneficial to oocyte development. In this work, we compared three protocols for OPU and in vitro production (IVP) of embryos, in Nellore cattle. Nellore cows (n = 18) were randomly allocated in three groups: Group 1 (OPU), Group 2 [Follicle stimulating hormone (FSH) and OPU] and Group 3 (FSH deprivation and OPU). Three OPUs were performed, and the animals were switched to a different group each time (crossover), in such a way that at the end of the experiment all cows received the 3 protocols. At random stage of the oestrous cycle (D‐2), all follicles ≥ 6 mm were aspirated to induce a new follicular wave 2 days afterwards (D0). In Group 1, OPU was performed on D2 and oocytes were processed to IVP. In Group 2, starting on D0, cows were superstimulated (FSH, Folltropin®, 30 mg administered daily, i.m., during three consecutive days, total dose = 180 mg), and 6 h after the last FSH dose, they received exogenous luteinizing hormone (LH) (12.5 mg, i.m., Lutropin®, D3). The OPU was performed 6 h after LH administration, i.e. 12 h after the last dose of FSH. Animals in Group 3 received the same treatment as those in Group 2, except that LH was administered 42 h after the last dose of FSH, and OPU occurred 6 h later. Therefore, in this group, follicles were deprived of FSH at 48 h. Both cleavage and blastocyst rates were similar (p > 0.05, anova ) among oocytes from Groups 1, 2 and 3, respectively: 77.4% (144/185) and 42.70% (79/185); 75.54% (105/139) and 31.65% (44/139); 63.52% (101/159) and 33.33% (53/159). However, hatched blastocyst rate was higher (p < 0.01) in Group 1 (30.27%, 56/185) when compared with Group 2 (11.51%, 16/139) or 3 (15.72%, 25/159). It is concluded that, contrary to previous work on European breeds (Bos taurus), ovarian superstimulation associated with deprivation of FSH and OPU (Group 3) did not increase IVP of Nellore embryos (Bos indicus). On the contrary, the highest hatched blastocyst rates were observed in oocytes from non‐superstimulated cows.  相似文献   

14.
The aim of this retrospective study was to compare the number of follicles, cumulus oocyte complexes (COCs) and cultured In Vitro Produced (IVP) embryos obtained from 1396 non‐stimulated Ovum Pick‐up (OPU) sessions on 81 donor animals in a twice weekly OPU scheme. Results were obtained from 640 sessions following FSH‐LH superstimulation, on 112 donors subjected to OPU once every 2 weeks. The stimulation protocol started with the insertion of an ear implant containing 3 mg norgestomet (Crestar, Intervet, Belgium) 8 days before puncture (day ?8). The dominant follicle was ablated by ultrasound‐guided follicle puncture on day ?6. On day ?3 and day ?2, cows were injected with FSH (Ovagen, ICP) twice daily (8 am to 8 pm ), i.e. a total dose of 160 μg FSH and 40 μg LG per donor per stimulation cycle. Animals were punctured 48 h after the last FSH injection (day 0). Progesterone implants were removed the next day. Stimulated donor cows were treated with this protocol at 14‐day intervals. Follicles were visualized with a Dynamic Imaging ultrasound scanner, equipped with a 6.5 MHz sectorial probe. Follicles were punctured with 55 cm long, 18 gauge needles at an aspiration pressure corresponding to a flow rate of 15 ml/min. Cumulus oocyte complexes were recovered and processed in a routine IVF set‐up. Results demonstrate that, expressed per session, FSH stimulation prior to OPU increases production efficiency with significantly more follicles punctured and oocytes retrieved. However, when overall results during comparable 2‐week periods are considered (four non‐stimulated sessions vs one stimulated), more follicles are punctured and more oocytes are retrieved using the non‐stimulated protocol. No significant differences in the number of cultured embryos could be detected, indicating that FSH/LH stimulation prior to OPU might have a positive effect on in vitro oocyte developmental competence as more embryos are cultured with less, presumably better‐quality, oocytes.  相似文献   

15.
Two experiments were conducted to evaluate oocyte collection rates and in vitro nuclear maturation rates of equine oocytes obtained during diestrus and pregnancy, and to compare these rates with maturation rates in oocytes derived from preovulatory follicles. In Experiment I, transvaginal ultrasound-guided aspiration of follicle was performed during estrus and diestrus in 14 mares over four consecutive cycles. Follicular aspirations during estrus were performed 24 to 27 hours after injection of 2500 IU of hCG given when the largest follicle reached 35 mm in diameter. Oocyte recovery rate from preovulatory follicles was 51% (33/65) in 49 aspiration sessions. Cumulus-oocyte complexes from preovulatory follicles were cultured for 12-15 hours in TCM199 + 10% (NCS) at 38.5°C in 5% CO2 in air, and 22/33 (67%) were in metaphase II. During diestrus, mares were treated (Group I) or not treated (Group II) daily with equine pituitary extract (EPE) during alternate cycles from days 1 to 14 after the preovulatory aspiration. Diestrous follicles were aspirated when four or more follicles greater than 12 mm in diameter were present. EPE had no effect on the number of follicles that developed during estrus or diestrus (p>0.05). Oocytes were recovered from 119 of 383 diestrous follicles (31 %)in 75 sessions. There was no difference in recovery rates between Groups I and II (p>0.05). Maturation rates for oocytes collected during diestrus, after 42 hours of culture in TCM 199 + 1 μg/ml of Estradiol and lnl/ml of EPE, were not significantly different (p>0.05) between Groups I and II (49% vs. 53%). In Experiment II, mares between 50 and 85 days of pregnancy were used as oocyte donors. The oocyte recovery rate was 53% (66/125). After in vitro maturation for 40 hours (compact COC) or 15 hours (expanded COC), 22% (7/32) and 22% (7/32), respectively, of the oocytes were in metaphase II. It was concluded that: 1) Preovulatory follicles yield a higher percentage of oocytes with a higher rate of maturation to metaphase II than follicles of diestrus and pregnant mares. 2) Diestrous follicles yielded fewer oocytes than follicles of pregnant mares but with a higher percentage of oocyte maturation. Further studies are necessary to determine if oocytes recovered from diestrous follicles and matured in vitro can be fertilized successfully.  相似文献   

16.
Experiments were conducted to determine the effects of lamb age, frequency of follicular aspirations, and hormone stimulation by fixed or variable FSH dose, on the number of collected oocytes and their maturational competence. In trial 1, the characteristics of follicular population (number and diameter of follicles) were studied in 40 lambs which were slaughtered at the age of 30 days (S1), 42 days (S2), 60 days (S3) and 5–6 months (S4), each n = 10. In trial 2, 27 lambs were divided into four groups. group MF lambs (n = 6) had follicular aspiration (OPU) in four monthly intervals commencing from the age of 8–9 weeks (sessions MF1, MF2, MF3 and MF4). In groups SF2, SF3 and SF4 (each n = 6), OPU was conducted once during the 12–13, 16–17 and 20–21 week of age, respectively. Ovarian stimulation was conducted with fixed FSH dose (3.52 mg/animal). In trial 3, 10 lambs (group MV) were treated as those of group MF apart from the FSH dose, which was administered according to the body weight in a dose of 0.27 mg/kg. The number and the size of follicles, the number and the quality of collected oocytes and the maturational competence of the oocytes were compared between and within groups. In trial 1, the total number and the number of small follicles were greater in groups S1 and S2 compared with those of S3 and S4 (p < 0.01). Similarly, the follicular population was greater in group MF1 than in group SF3 (p < 0.01). In sessions MF2, MF3, MV2, MV3 and MV4, more oocytes were collected in comparison with those from the respective once‐aspirated age mates (groups SF2, SF3 and SF4). In total, more (p = 0.02) oocytes per donor were collected from group MV (15.2 ± 5.5) than from group MF (9.0 ± 3.2). An absolute maturational failure was observed in oocytes collected from groups SF2 and SF3. Maturational competence varied between 16.7% and 58.3% (p = 0.017) among sessions of group MF, but it was more uniform among sessions of group MV (range 12.5–42.9%, p > 0.05). Our results indicate that firstly, the number and the quality of harvested oocytes from juvenile lambs can be much improved if follicular stimulation regime is adjusted to the body weight. Secondly, in terms of follicular population and oocyte quality, 3 and 4‐month‐old lambs are naturally bad oocyte donors, but this characteristic can be reversed by a previous follicular ablation.  相似文献   

17.
将8头荷期坦供体奶牛随机分为4组,每组2头,分别按优势卵泡+注射促卵泡素(FSH)、优势卵泡+不注射FSH、去优势卵泡+注射FSH、去优势卵泡+不注射FSH处理供体牛,处理后第4天应用B型超声波导引活体采卵,试验重复10次。结果显示:注射FSH前去优势卵泡对其可采卵泡数、回收卵母细胞数和可用卵母细胞数均有显著提高(P0.05),但不注射FSH时去优势卵泡对提高采卵效率无显著效果;活体采卵对供体牛卵巢虽造成一定损伤,但停止采卵2~3个月后均能恢复正常发情、配种后能正常妊娠,对供体牛繁殖性能无显著影响。  相似文献   

18.
Six oocytes were aspirated from preovulatory follicles of 18 mares via surgical exposure of the ovary (5/13) or percutaneous needle aspiration (1/5). Surgical transfer of the oocytes into the oviduct of mares previously inseminated with 500 × 106 progressively motile spermatozoa resulted in no embryos recovered by uterine lavage 8 days afteroocyte transfer. Corpora luteal function and formation from aspirated follicles appeared normal, based on progesterone analysis, ultrasound morphological assessment and cyclic behavioral changes.  相似文献   

19.
In order to provide a consistent system for laboratory production of embryos, the characteristics of immature camel oocyte must first be described. The objective of this study was to define ultrastructural features of immature camel oocyte. Ovaries were obtained from camels at a local abattoir, and then transported to the laboratory within 2 h. Camelus cumulus oocyte complexes (COCs) were aspirated from 2-6 mm follicles using a 22-gauge needle. Excellent and good quality COCs were selected and prepared for transmission electron microscopy study using a cavity slide. The fine structure of camel oocyte is morphologically similar to that of other mammalian oocytes. However, some minor differences exist between COC of camel and other mammalian species. Different size and shape of membrane-bound vesicles, lipid droplet, mitochondria and cortical granules were distributed throughout the ooplasm. Discrete or in association with endoplasmic reticulum, Golgi complexes were observed in the periphery of the oocytes. The majority of the oocytes were in the germinal vesicle stage.  相似文献   

20.
This study examined the effect of treating mares with equine pituitary extract (EPE) in combination with human chorionic gonadotropin (hCG; EPE/hCG) on the recovery rate of immature oocytes by ovum pick-up (OPU) and on oocyte morphology. Ten mares were subjected to each of two treatments in a random sequence: superstimulated with EPE (25 mg, twice daily) and treated with hCG (2,500 IU) or control (no exogenous treatment). The cytoplasmic morphology of oocytes recovered was evaluated through transmission electron microscopy. Follicular fluid was collected at aspiration for progesterone analysis, which was performed by radioimmunoassay. The EPE/hCG did not increase the oocyte recovery rate from immature follicles when compared with the controls (15.5% and 16.7%, respectively). A significantly higher oocyte recovery rate per mare was observed (70% versus 50%). However, precocious granulosa cell expansion was observed with EPE/hCG treatment in contrast to the control (64.4% and 33% of follicles with expanded cumulus, respectively), and increased intrafollicular progesterone concentration was also seen (158.80 ng/mL versus 82.05 ng/mL). The ultrastructural analysis of oocytes from both groups showed morphologic features related to immaturity. Numerous vesicles containing cortical granules were found, distributed in clusters into the cytoplasm, and junctional complexes were still seen between oocyte and granulosa cells. In conclusion, EPE/hCG treatment induced some follicular modifications, but the recovery rate was not increased. All oocytes examined presented signs of immaturity.  相似文献   

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