Effects of progestagens on follicular growth and oocyte developmental competence in FSH-treated ewes

Previous research has reported evidence for negative effects of progestagens on follicular growth and oocyte competence. In the present study, negative effects of progestagens on follicular growth and oocyte developmental competence were assessed. During the breeding season, 20 Sarda ewes were treated with two doses of cloprostenol, 10 days apart, to assure the presence of a corpus luteum (CL). On day 5 after the second cloprostenol dose, 10 ewes were treated with a progestagen sponge while 10 females remained untreated. Starting on day 7 after the second cloprostenol dose, all the ewes were treated with 6 equal doses of 24 I.U. of FSH (Ovagen™, ICP, NZ), every 12 h. The number of follicles ≥2 mm in diameter increased (P < 0.0005) in all the ewes from 24 h before to 60 h after the first FSH dose (from 12.8 ± 1.1 to 23.4 ± 1.3 in treated and from 12 ± 0.6 to 22 ± 1.2 in untreated ewes, n.s.). There were no significant differences in follicle dynamics between groups, but concentrations of estradiol in control ewes were higher than in the progestagen group (P < 0.05). Twelve hours after the last FSH dose, oocytes were collected by ovum pick-up. Recovery rates were lower for progestagen-treated ewes (71.1 versus 83%; P < 0.001). After IVP procedure, cleavage rate was also lower in the progestagen group (39.1 versus 82.6%; P < 0.001). Furthermore, blastocysts output revealed that oocyte developmental competence was lower in progestagen group (17.3 versus 30.4%; P = 0.245), although differences were not significant. These results suggest deleterious effects from progestagen on oocyte developmental competence and set the basis for new protocols for in vitro embryo production.     

Effect of feline interferon-omega on the survival time and quality of life of cats with feline infectious peritonitis

BACKGROUND: There is no therapy with proven efficacy to treat cats with feline infectious peritonitis (FIP). HYPOTHESIS: Feline interferon-omega (FeIFN-omega) prolongs survival time and increases quality of life in cats with FIP. ANIMALS: Thirty-seven privately owned cats were subjects of this study. METHODS: The study was performed as a placebo-controlled double-blind trial. Feline infectious peritonitis was confirmed by histology or immunostaining of feline coronavirus (FCoV) antigen in effusion or tissue macrophages or both. The cats were randomly selected for treatment with either FeIFN-omega or a placebo. All cats received adjunctive treatment with glucocorticoids and antibiotics and passive immunization with Feliserin. RESULTS: There was no statistically significant difference in the survival time of cats treated with FeIFN-omega versus placebo or in any other variable evaluated (with the exception of the lymphocyte count). The cats survived between 3 and 200 days (median, 9 days). There was only 1 long-term survivor (> 3 months), and the cat was in the FeIFN-omega group. CONCLUSION AND CLINICAL RELEVANCE: No effect of FeIFN-omega on survival time or quality of life could be demonstrated in this study.     

Effect of open pulled straw (OPS) vitrification on the fertilisation rate and developmental competence of porcine oocytes

Freezing technologies are very important to preserve gametes and embryos of animals with a good pedigree or those having high genetic value. The aim of this work was to compare immature and in vitro matured porcine oocytes regarding their morphology and ability to be fertilised after vitrification by the open pulled straw (OPS) method. In four experiments 830 oocytes were examined. To investigate the effect of cumulus cells on oocyte survival after OPS vitrification, both denuded and cumulus-enclosed oocytes were vitrified at the germinal vesicle (GV) stage, then after vitrification they were matured in vitro. Besides, in vitro matured oocytes surrounded with a cumulus and those without a cumulus were also vitrified. The survival of oocytes was evaluated by their morphology. After in vitro fertilisation the rates of oocytes penetrated by spermatozoa were compared. Our results suggest that the vitrification/warming procedure is the most effective in cumulus-enclosed oocytes (22.35 +/- 1.75%). There was no difference between the order of maturation and vitrification in cumulus-enclosed oocytes, which suggests the importance of cumulus cells in protecting the viability of oocytes during cryopreservation.     

Effect of trans-10 cis-12 conjugated linoleic acid on bovine oocyte competence and fatty acid composition

The reproductive performance of dairy cows may be improved by feeding conjugated linoleic acid (CLA) supplements during early lactation. The mechanism of action of t10,c12 CLA is not clearly known. Our objective was to investigate the effect of t10,c12 CLA on oocyte maturation and lipid composition of cumulus oocyte complexes (COC). The developmental potential of oocytes incubated in in vitro maturation (IVM) medium supplemented with t10,c12 CLA to the blastocyst stage and embryo quality were also assessed. In experiment 1, abattoir-derived oocytes were matured in TCM199 + 10% serum supplemented with 100 μM t10,c12 CLA (t10,c12 CLA n = 672) or without it (control n = 672). Mature oocytes were either stained for chromatin configuration or inseminated and cultured for embryo development assessment. In experiment 2, COC and IVM culture media were subjected to fatty acid (FA) analysis prior and after maturation with t10,c12 CLA or without it (control). Total lipids and FA profiles in oocytes, cumulus cells and culture media were determined by gas chromatography. t10,c12 CLA supplementation to IVM medium improved (p = 0.05) embryo quality evaluated morphologically. This effect was associated with t10,c12 CLA presence (3.1 ± 0.7%, p = 0.04) and lower levels of arachidonic acid in FA profile of t10,c12 CLA mature oocytes (immature oocytes = 4.4 ± 1.9%, t10,c12 CLA mature oocytes = 1.0 ± 0.7%, p = 0.05). Differences in myristic and eicotrienoic acids, saturated and unsaturated FA concentrations between oocytes and cumulus cells were detected (p ≤ 0.05). In conclusion, the presence of t10,c12 CLA during maturation interfered on lipid metabolism improving bovine oocyte competence to develop into higher quality embryos.     

牛卵母细胞体外受精技术

胚胎移植技术作为种公牛的引进、高产奶牛和优质肉牛扩繁的技术手段,在生产中发挥了良好作用.但是,胚胎移植技术在应用过程中均存在胚胎资源短缺、胚胎价格过高的问题.     

Chronological changes of bovine follicular oocyte maturation in vitro

Chronological changes of bovine follicular cumulus-oocyte-complexesi were studied after in vitro maturation over a period of 48 h. According to their thickness and compactness of cumulus investments they were classified into 4 groups and cultured in enriched Ham’s F-10 medium with or without human chorionic gonadotrophin (hCG) and estradiolbenzoate (EB) for 0, 6, 12, 18, 21, 24, 27, 30 and 48 h. Representative samples were taken at each time interval for evaluation of nuclear maturation stages, ooplasm quality and size of the peri vitelline space (PVS). The results showed that oocyte nuclear breakdown (ONBD) required 6 to 12 h culture, and the peak of the first polar abstriction occurred at 24 h. The culture period required for ONBD and abstraction of the first polar body were related to the thickness and compactness of cumulus investments with and approximately 6 h delay in heavily compacted complexes. Ooplasm quality evaluation failed to show a clear trend, but the PVS increased in size from 0 h to 30 h and then, retracted again from 30 to 48 h. The overall maturation rate in the presence of hCG and EB was 79.1 %, and a substantial proportion (68.8 %) of nude or partially covered oocytes reached metaphase II stage. In the presence of hCG and EB no block at either metaphase I or at anaphase-telophase I was observed. In the absence of hCG and EB the percentage of oocytes reaching metaphase II was much lower (48.6%) in comparison with oocytes matured in the presence of these hormones (79.1 %). It was concluded a very high proportion of slaughterhouse oocytes could be matured in vitro and that the cumulus investments and addition of certain hormones affected the maturation rate.     

Effect of different mechanical isolation techniques on developmental competence and survival of buffalo ovarian preantral follicles

The present study aimed to analyze different methods of mechanical isolation of buffalo preantral follicles (Experiment I), in vitro culture of isolated follicles in different groups of culture medium over collagen gel matrix (Experiment II) and subsequent in vitro development and survival of isolated preantral follicles (PFs) (Experiment III). In Experiment I, ovarian cortical pieces were separated and PFs isolated by different mechanical methods. In Experiment II, isolated follicles were divided into three groups and cultured in TCM-199 having 10% FBS, 1% ITS, and 20 ng/ml EGF (Group A, control), addition of 0.5 μg/ml FSH (Group B) or FSH + 100 ng/ml IGF-I (Group C). Follicles were incubated at 38.5 °C in 5% CO₂ with maximum humidity. In Experiment III, based on the outcome of Experiment I and II, PFs were cultured from those isolation method and treatment group, showing better growth and developmental pattern to analyze the impact of growth factors on in vitro growth of follicles in long term culture. It was found that micro-dissected PFs showed higher survival rate and growth after 15 days of culture compared to PFs isolated by other methods. Follicles cultured with FSH + IGF-1 on collagen gel matrix, showed significantly (P < 0.05) higher survival rate and mean diameter of follicles on day 15 of culture compared to control. In summary, it has been shown that isolation of follicles by micro-dissection has advantages over other methods, being relatively simple, inexpensive and less harmful to follicles. Micro-dissected buffalo PFs maintained the architecture, showed antrum formation in presence of FSH and IGF-I over the collagen gel matrix.     

能量对家畜卵泡发育的影响及调控

家畜卵泡发育是被众多因素精确调控的生理过程。日粮能量可能通过不同的机制影响卵泡发育。胰岛素、瘦素和IGF-Ⅰ可能作为代谢信号而介导能量对卵泡发育的影响,即通过下丘脑-垂体-卵巢轴调节促性腺激素释放或直接作用于卵巢。各种代谢信号之间的相互作用也是其介导作用发挥的一个重要方面。     

玻璃化冷冻预处理方法对牛卵巢颗粒细胞的影响

为了研究不同玻璃化冷冻预处理对牛卵巢颗粒细胞培养的影响,试验采用4种不同的玻璃化冷冻预处理方法(①将冷冻保存管直接投入液氮罐中;②将冷冻保存管先置于4℃冰箱15 min,然后投入液氮罐中;③将冷冻保存管先置于-20℃冰箱10 min,然后投入液氮罐中;④将冷冻保存管先置于4℃冰箱15 min,再置于-20℃冰箱10 min,然后投入液氮罐中)处理牛卵巢颗粒细胞,解冻后检测细胞活力和传代时间。结果表明:解冻后4种方法的细胞活力分别为72%、74%、79%和86%;传代时间分别为76 h、73 h、67 h和62 h;方法④的细胞活力和传代时间与其他组比较差异显著(P<0.05)。说明玻璃化冷冻预处理方法有利于复苏后的牛卵巢颗粒细胞的进一步培养。     

Effect of a single injection of progesterone on ovarian follicular cysts in lactating dairy cows

The objective of this experiment was to evaluate the effect of a single injection of progesterone on the lifespan of ovarian follicular cysts and to examine the fate of follicles that mature following treatment. Lactating Holstein and Jersey cows with ovarian follicular cysts were identified by rectal palpation. The ovaries of cystic cows were then examined by transrectal ultrasonography three times weekly to monitor formation of new follicular cysts. Cows with newly formed follicular cysts were treated either with a single injection of progesterone (200 mg, IM, n = 11) or corn oil vehicle (n = 7). Venous blood samples were collected daily for quantification of progesterone. Blood sampling and ultrasonography continued until ovulation or a new follicular cyst formed. Treatment reduced the lifespan of the cyst by 12 days, from 29.8 +/- 2.3 days in control cows to 17.2 +/- 1.8 days in progesterone-treated cows (P = 0.01). Progesterone treatment also tended to alter the frequency of subsequent follicular events. Ovulation occurred in 4/11 cows that were treated with progesterone whereas none of the vehicle treated cows ovulated (P = 0.07). In conclusion, a single injection of 200mg of progesterone, administered early in the life of an ovarian follicular cyst, shortened its lifespan and in some cases was followed by ovulation of a new follicle.     

玻璃化冷冻前平衡及解冻后恢复处理对卵母细胞质量的影响

以小鼠为动物模型，研究了玻璃化冷冻前的平衡和解冻后的恢复处理对卵母细胞质量的影响程度。结果表明：1）无论是受精卵还是卵母细胞，其集聚FDA（乙酰荧光素）没有明显差异，可以用其检测卵母细胞活力；2）在产销认中添加CB组的卵母细胞细胞膜完整率与CB组没有明显差异；但是明显低于对照组。3）恢复处理在无CB时对卵母的激活作用并不明显，虽然活化率有一定的提高，但是添加CB组的卵母细胞激活率却明显增加，但是激活的类型多是排出第二极体，形成原核的比率很低。     

Survey of canine and feline follicular tumours and tumour-like lesions in central Italy

Between January 1985 and June 1997, a total of 4005 skin biopsies were received from dogs and 898 from cats. Follicular tumours and tumour-like lesions together represented 10.4 per cent and 8.1 per cent of all skin tumours in the dog and cat, respectively. The prevalence of tumour-like lesions, such as follicular and dermoid cysts, dilated pore and focal adnexal dysplasia, was highest, representing 41.2 per cent and 68 per cent of all follicular lesions in dogs and cats, respectively. In the dog, follicular tumours were distributed as follows: trichoblastoma (25.6 per cent of all tumours and tumour-like lesions), infundibular keratinising acanthoma, (14 per cent), pilomatricoma (13 per cent), trichoepithelioma (3.9 per cent) and tricholemmoma (2.3 per cent). In the cat, the distribution was 26 per cent for trichoblastoma, 4 per cent for trichoepithelioma and 2 per cent for pilomatricoma. Tumour-like lesions and infundibular keratinising acanthoma in the dog were mostly located on the trunk, trichoblastoma and tricholemmoma on the head, and pilomatricoma on the neck. In the cat, both tumour-like lesions and trichoblastoma were frequently present on the neck and head.     

玻璃化冷冻对小鼠卵母细胞透明带的影响

本文旨在通过研究玻璃化冷冻小鼠卵母细胞透明带超微结构、透明带厚度(ZPT)、厚度变量(ZPTV)及双折射分值(ZPB)的影响,分析三者间的相关性,探求玻璃化冷冻液的最佳配方。选用4组应用最广泛的冷冻液配方对小鼠卵母细胞进行处理,与未处理的卵母细胞比较存活率、受精率、卵裂率和囊胚率,选出最适冷冻液配方。以新鲜卵母细胞为对照组,进行冷冻程序但并没有进行实际冷冻的卵母细胞为处理组,进行玻璃化冷冻复苏的卵母细胞为冷冻组,扫描电镜观察各组的透明带超微结构变化;检测3组细胞的ZPT和ZPB,并对ZPT和ZPB、ZPTV和ZPB之间相关性进行分析。结果发现HM+7.5%(DMSO+EG),HM+15%(DMSO+EG)+0.5 mol/L Su冷冻液组对卵母细胞发育潜力影响较小。玻璃化冷冻对ZPT(6.05±0.10μm vs 5.77±0.60μm)和ZPB(0.30±0.38 vs 1.22±0.21)有显著影响(P<0.05),且ZPT与ZPB呈负相关(r=-0.299)(P<0.05)。扫描电镜发现玻璃化冷冻造成卵母细胞透明带呈熔融状,表面凹凸不平,窗孔基本不可见,甚至出现透明带部分脱落的情况;冷冻组透明带超微结构的正常率较对照组和处理组下降,其中粗ZP(44.9%对92.9%和84.8%)(P<0.01)和光滑ZP(31.0%对7.4%和9.1%)(P<0.01)。结果表明,玻璃化冷冻影响卵母细胞的发育潜能,低温对透明带的超微结构有较大的负面影响。     

Relationship between ovarian weight and follicular population in heifers

The size of the ovary varies substantially among cattle. This variation may influence the potential of the ovary to produce follicles. In the present study, we examined whether a relationship exists between the weight of the ovary and the number of antral follicles >or=1 mm. Paired ovaries were obtained from Holstein x Japanese Black F1 heifers. Follicles were classified into three size categories (small: 1.0-<5.0 mm, medium: 5.0-<8.5 mm and large: >or=8.5 mm), and the number of follicles in each category was recorded. Large variations in the weight of ovaries and the number of follicles were observed among animals. Significant positive correlations (r>or=0.4, P<0.001) were found between the weight of intact ovaries and the number of follicles in all three categories for the ovary contralateral to CL (OCC) and in the small follicles for the ovary ipsilateral to CL (OIC). Significant positive correlations (r>0.4, P<0.0001) were also observed between the weight of ovaries devoid of CL and follicles and the number of small and medium follicles in both OIC and OCC, indicating that the correlation is not due to the increase in ovarian weight associated with the increase in follicular number. Paired ovaries contained a similar number of small and medium follicles, and significant positive correlations were observed between them (r>0.6, P<0.0001). There were significant positive correlations between the weight of OCC and the number of small and medium follicles in paired ovaries (r>0.4, P<0.0001). These results suggest that 1) the weight of an ovary reflects the potential of the ovary to produce antral follicles, and 2) a rough estimation of follicular population might be possible by using the weight of the ovary contralateral to CL in heifers.     

Effect of time of oocyte collection and site of insemination on oocyte transfer in mares

The objective of the study was to compare embryo development rates after transfer of oocytes collected 22 or 33 h after hCG injection into recipients inseminated within the uterus or the oviduct. Oocytes were collected at approximately 22 or 33 h after hCG injections and incubated for approximately 16 or 1.5 h, respectively, before transfer. Intrauterine inseminations using 1 x 10(9) progressively motile sperm were done approximately 12 h before and 2 h after transfer. For intraoviductal inseminations (gamete intrafallopian transfer [GIFT]), semen was centrifuged through a Percoll gradient, and 200,000 progressively motile sperm were transferred with oocytes into the oviduct. Time of oocyte collection (22 or 33 h) after hCG injection did not affect embryo development rates (17/25, 68%, vs 12/23, 52%, respectively; P = 0.40). When results from oocyte collections at 22 and 33 h after hCG were combined, oocyte transfer with intraoviductal vs intrauterine insemination resulted in similar (P = 0.70) embryo development rates (12/22, 55%, and 17/26, 65%, respectively). However, the interaction between time of oocyte collection and site of insemination tended to be significant (P = 0.09), suggesting that GIFT using oocytes collected at 33 h after hCG may not be as effective as using oocytes collected at 22 h after hCG. Because intraoviductal insemination requires a low number of sperm, GIFT could be used in cases of male subfertility, frozen semen, or sexed sperm.     

Effect of unilateral ovariectomy and injection of bovine follicular fluid on gonadotropin secretion and compensatory ovarian hypertrophy in prepuberal heifers

A study was conducted to determine whether compensatory ovarian hypertrophy occurred in prepuberal heifers, and if so, could compensatory ovarian hypertrophy be prevented by injecting charcoal-extracted bovine follicular fluid (CFF). An additional objective was to determine the effect of CFF injections on follicle stimulating hormone (FSH) and luteinizing hormone (LH) concentrations following unilateral ovariectomy (ULO). Thirty-one prepuberal Angus, Hereford, and Angus X Hereford heifers were assigned by weight and breed to the following groups: Control-saline injections (C), unilateral ovariectomy-saline injections (ULO) and unilateral ovariectomy-CFF injections (ULO-CFF). In group C, a sham operation was performed on d 0 and both ovaries were removed on d 7. In groups ULO and ULO-CFF, either the right or left ovary was removed on d 0 and the remaining ovary was removed on d 7. From d 0 to 7, each heifer received an injection (iv, 7 ml/injection) of saline or CFF every 12 h. Jugular blood samples were collected immediately before each saline injection from d 0 to 7 and analyzed for FSH and LH. Unilateral ovariectomy increased ovarian (P less than .08) and follicular fluid (P less than .05) weights compared with groups C and ULO-CFF; however, lyophilized ovarian weight was similar (P greater than .05) for all groups. Injections of CFF blocked the increase in ovarian and follicular fluid weight following ULO. The total number of visible follicles per ovary was similar for all groups; however in the ULO group there was an increase (P less than .01) in those 5 to 6 mm and greater than or equal to 9 mm.(ABSTRACT TRUNCATED AT 250 WORDS)