Effects of Cholesterol on Progesterone Production by Goat Luteal Cell Subpopulations at Two Different Stages of the Luteal Phase

The aim of the present study was to evaluate the effects of cholesterol on progesterone production during long‐term culturing of luteal cell subpopulations at early and late luteal stages of the goat corpora lutea. Corpora lutea were collected from Angora goats on days 5 and 15 of the oestrous cycle. Luteal cells were isolated by collagenase digestion. The cells were separated into two distinct subpopulations by Percoll density‐gradient centrifugation. Both subpopulations of luteal cells staining positively for 3β‐HSD activities (5 × 10⁴ cell/well) were cultured with or without 22(R)‐hydroxycholesterol (22R‐HC) in serum‐free culture medium for periods of up to 7 days. Cells were incubated with serum (10%) for the first 18 h of incubation followed by serum‐free medium. Cell treatment (10 and 20 μg/ml) was performed on days 1, 3 and 5. Treatment of cells with both concentrations of 22R‐HC resulted in significant (p < 0.01) and dose‐dependent stimulation (p > 0.05) on progesterone production in both fractions of cells throughout 7 days of incubation. Treatment of the cells with cholesterol resulted in 2.5‐ and 9.0‐fold increases in progesterone accumulation on day 3 of incubation. Steroid production was maintained throughout the incubations when cells are incubated in serum‐free media treated with cholesterol and ITS premix. Cells collected from higher density of percoll layers produced 2.82 and 2.32 times more progesterone, in comparison to the lover density percoll layer, on days 5 and 15 of the oestrous cycle in untreated cell groups, respectively. Progesterone accumulation was decreased as incubation time advanced in all groups of untreated cells. These results demonstrated that goat luteal cell subpopulations secrete substantial amounts of progesterone in response to cholesterol treatment at least for 7 days, and cholesterol is required as progesterone precursor for maintaining a high‐level steroidogenesis during long‐life culturing of both cell subpopulations.     

排卵延迟奶牛发情后不同阶段B超黄体相关指标变化规律分析

为研究排卵延迟奶牛发情配种后黄体的变化规律及其差异性,本研究在临床症状观察、直肠检查和B超辅助检查诊断的基础上,选取8头排卵延迟奶牛(排卵延迟组)、5头健康正常发情奶牛(对照组),应用B超对黄体的相关指标(直径、面积、周长和体积)进行了测量及分析,对获得的不同阶段典型黄体声像图进行了描述。结果显示,与对照组相比,排卵延迟组与对照组奶牛黄体的直径、面积、周长和体积在发情配种后7~15 d差异均不显著(P>0.05)。在发情配种后第9和13天排卵延迟组奶牛黄体相关指标(直径、体积、面积和周长)大于对照组奶牛,而在第11天小于对照组奶牛。结果表明,排卵延迟奶牛发情后不同阶段B超黄体直径、面积、周长和体积与正常发情排卵后的相应指标没有显著差别。     

Analysis of B-ultrasound Luteal Variation Related Indicators of Delayed Ovulation Dairy Cows after Estrus at Different Stages

The study was aimed to observe the variation and differences of corpus luteum of delayed ovulation cows after estrus.Based on the observation of clinical symptoms, rectal examination and diagnosis of B-ultrasound scanning, 8 delayed ovulation dairy cows with the variation (delayed ovulation group) and 5 healthy and normal dairy cows with estrus (control group) were selected, and the related indicators of the corpus luteum (diameter, area, perimeter and volume) were measured by B-ultrasound scanning.These luteal indicators were compared and analyzed further, and the luteal typical sonography obtained of the different stages were described.The results showed that the diameter, area, perimeter and volume during 7 to 15 d after estrus of the corpus luteum had no significant differences between delayed ovulation group and control group (P>0.05).However, on the 9th and 13th day after estrus, the luteal relevant indicators of delayed ovulation group cows (diameter, volume, area and perimeter) were greater than that of control group cows and less than normal dairy cows on the 11th day.There were no significant differences in diameter, area, circumference and volume of corpus luteum at the different stages after estrus between the delayed ovulation dairy cows and the normal ones.     

不同收获期玉米青贮结构性碳水化合物对奶牛瘤胃发酵的影响

用3头装有永久性瘘管的中国荷斯坦牛,采用3×3拉丁方设计对蜡熟期、乳熟期和乳熟前期的玉米青贮的NDF和ADF进行了瘤胃降解率及其日粮对瘤胃发酵内环境的影响进行了研究。结果表明,乳熟期玉米青贮NDF在瘤胃中的有效降解率(30．77％)分别高于蜡熟期(28．59％)和乳熟前期(26．71％,P<0．05),蜡熟期和乳熟前期之间差异不显著。蜡熟期(25．73％)和乳熟前期(25．23％)玉米青贮ADF有效降解率均高于乳熟期(23．39％,P<0．05),但蜡熟期和乳前熟期之间没有明显差异。饲喂乳熟前期、乳熟期、蜡熟期玉米青贮组成的日粮的奶牛瘤胃液pH值变动范围分别为6．37～7．27,6．37～6．97,6．33～7．27;NH3-N浓度的平均值分别为33．32 mg／dl, 38．87 mg／dl,34．53 mg／dl;乙酸、丙酸和丁酸浓度分别为49．48,12．58,4．54 mmol／L、49．83,10．28,6．41 mmol／L和44．24,9．32,5．43 mmol／L。表明三组牛瘤胃pH值、NH3-N和VFA浓度没有显著性差异(P>0．05)。     

Histological Characteristics and Steroid Concentration of Ovarian Follicles at Different Stages of Development in Pregnant and Non-pregnant Dairy Cows

The aim of the study was to investigate the histological characteristics and steroid concentrations in follicular fluid of different populations of follicles at different stages of development, during pregnancy and the oestrous cycle in cows. Follicles from ovaries collected at a slaughterhouse were allocated into three size categories (small, 2–5.9 mm; medium, 6–13.9 mm; and large, 14–20 mm) in pregnant and non-pregnant cows. Slices were stained with HE and PAS for histological analysis. Follicular fluid was pooled according to size and pregnancy status and estradiol, testosterone and progesterone concentrations in follicular fluid were determined by RIA. Characteristics of healthy follicles did not differ, regardless of follicle size or pregnancy status. Total histological atresia was significantly higher in pregnant cows than in non-pregnant cows (p < 0.05). Estradiol increased and testosterone decreased significantly, while follicles increased in size, in both non-pregnant and pregnant cows (p < 0.05). Nonpregnant cows had the highest estradiol values in follicles of all sizes. Medium and large follicles from pregnant cows showed the lowest testosterone concentration (p < 0.05). Progesterone levels increased with follicle size only in non-pregnant animals. In large follicles, progesterone concentration was significantly higher in non-pregnant cows than in pregnant cows (p < 0.05). Considering steroid concentration and histological findings, most large follicles might be atretic during pregnancy in cattle.     

Seasonal Changes in Luteal Progesterone Concentration and mRNA Expressions of Progesterone Synthesis-related Proteins in the Corpus Luteum of Mares

Although circulating progesterone (P(4)) levels tend to change with the season, little is known about the seasonal changes of P(4) synthesis-related proteins in the corpus luteum (CL) of mares. To examine these changes, seventy-four ovaries containing a CL were collected from Anglo-Norman mares at a local abattoir in Kumamoto, Japan (~N32°), five times during one year. The stages of the CLs were classified as early, mid and regressed by macroscopic observation of the CL and follicles. The mid CL, which had the highest P(4) concentration, was used to evaluate the seasonal changes in P(4) synthesis. The luteal P(4) concentration and mRNA expression of luteinizing hormone receptor (LHCGR) were lowest during early winter and highest during late winter. The mRNA expressions of steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage enzyme (P450scc) and 3β-hydroxysteroid dehydrogenase/Δ5-Δ4 isomerase (3β-HSD) were lowest during early winter and increased during late winter. These results suggest that P(4) synthesis in the CL is affected by the seasonal changes in the mRNA expressions of P(4) synthesis-related proteins in mares.     

The Effect of Progesterone and Oestradiol Benzoate on Fertility of Artificially Inseminated Repeat-breeder Dairy Cows during Summer

An experiment was conducted to examine the effect of progesterone (P(4)) and oestradiol benzoate (ODB) on fertility of repeat-breeder lactating dairy cows during summer. One hundred repeat-breeder lactating dairy cows were randomly allocated to four groups (Tr1, Tr2, Tr3 and C) in a study conducted at a private dairy farm. All cows were injected with 2 mg ODB (day 0), which were at random stages of their oestrous cycles. Cows in Tr1, Tr2 and Tr3 were administered with intravaginal progesterone-releasing devices (controlled internal drug-releasing, CIDR) at the time of ODB injection for 7 days and those in group C were untreated and served as controls. Following CIDR removal, all cows were given an intramuscular injection of 25 mg Prostaglandin (PGF(2 alpha)). Twenty-four hours after the PGF(2 alpha) injection, cows in Tr1, Tr2 and C groups were injected with 1 mg ODB. Cows in Tr3 group were injected with 10 microg gonadotropin-releasing hormone (GnRH) agonist 48 h after CIDR removal. Artificial insemination was performed between 24 and 30 h following the second ODB injection for cows in Tr1 group and at the time of GnRH injection for cows in Tr3 group. Cows in Tr2 and C groups were inseminated at detected oestrus. Plasma P(4) and oestradiol 17beta (E(2)) concentrations were determined for all cows daily from day 0 to day 9. Plasma concentrations of P(4) and E(2) among cows of groups Tr1, Tr2 and Tr3 were increased and reached maximum values within 48 h following administration and were greater (p < 0.001) than those of group C cows. The proportion of cows detected in oestrus based on P4 concentration on day 9 was 88%, 72%, 88% and 60% in groups Tr1, Tr2, Tr3 and C, respectively. Oestrous detection rate differed (p < 0.01) significantly between time-inseminated groups (Tr1 and Tr3) and those inseminated at observed oestrous (Tr2 and C) groups. Pregnancy rates based on ultrasonography performed on day 28 were 52%, 56%, 60% and 40%, and those based on rectal palpation on day 45 were 32%, 44%, 36% and 28% for Tr1, Tr2, Tr3 and C cows (p > 0.1), respectively. Whereas pregnancy rates for cows with four or more previous services in all groups (54.55%) were higher (p < 0.03) than those for cows with three previous services (29.49%). In pregnant cows, mean days from calving to the day of insemination were higher (p < 0.01) among cows with four or more previous services (204 +/- 8.0 days) than those with three previous services (157 +/- 6.0 days). Results indicate that treatment with a combination of ODB and CIDR in repeat-breeder dairy cows causes elevation in plasma concentrations of E(2) and P(4). Oestrous detection rate was better in cows that were primed with P(4) than those without P(4) priming. Cows with four or more previous services had significantly higher pregnancy rates than those with three previous services.     

宁夏地区不同生长阶段奶牛产污系数的对比分析

选择不同生长阶段的中国荷斯坦奶牛作为试验对象,采用全收粪尿法来研究不同生长阶段奶牛各种污染物特性及产污系数。结果表明:不同生长阶段奶牛体重与其采食量、粪尿产生量大致成二倍正相关关系;P、Cu、Zn元素都主要通过粪便排泄,其中Cu、Zn元素在尿液中含量甚微;单位体重粪或尿污染物产生量无论是从N、P还是Cu、Zn,成年泌乳奶牛对环境的贡献率都要高于育成期奶牛。     

Plasma Progesterone during the Luteal Phase and Pregnancy in Parturient and Barren Blue Fox Vixens

Abstract

The variation in progesterone secretion during the luteal phase and pregnancy in blue fox vixens was analyzed. Progesterone was measured in blood plasma once or twice a week using radioimmunoassay. The material was allocated into three groups; five mated, but barren, blue fox vixens, six mated vixens with implantation zonès in the uterus, but no cubs at parturition, and 26 normally parturient vixens. The progesterone profiles for the three different groups of females showed a steady increase in progesterone immediately after mating. Maximum values were observed on days 8–12 of pregnancy. Then the progesterone levels decreased gradually until delivery around day 52. The levels of progesterone were found to be significantly different (P<0.05) between non-pregnant and pregnant females from day 22 after mating. The plasma progesterone level seems to be affected by the presence of conceptuses.     

不同泌乳时期荷斯坦奶牛的行为观察

选用健康无病的奶牛12头（其中泌乳盛期4头,泌乳后期4头,干奶期4头）。按泌乳期分为3组。对各组奶牛逐头进行连续48h行为学观察。结果表明：牛夜间反刍时间（周期数）都比昼反刍时间（周期数）长（多）。泌乳盛期奶牛的总采食时间、反刍时间、采食咀嚼速度比泌乳后期和干奶期奶牛长（快）（P〉0．05）。饲后反刍开始时间泌乳后期最短,而干奶期最长。泌乳盛期奶牛饮水次数远比泌乳后期、干奶期多（P〈0．01）,排粪次数也多（P〈0．05）,排尿次数盛期牛与后期牛接近,但比干奶期牛多2次／d。干奶期牛的卧地时间最长,泌乳盛期牛次之,泌乳后期牛最短,但卧地次数泌乳盛期比后期、干奶期分别多4．13次和3、63次。这些行为的差异可为处于不同泌乳时期的荷斯坦奶牛提供适宜的饲养环境,制定科学的管理制度提供可靠的依据。     

泌乳盛期和泌乳后期奶牛的采食行为和日粮营养物质表观消化率的对比分析

试验旨在研究不同泌乳阶段奶牛饲喂相同日粮对采食行为和日粮营养物质表观消化率的影响。选择体重、胎次相近的8头健康泌乳牛,其中4头处于泌乳盛期(泌乳盛期组),4头处于泌乳后期(泌乳后期组),2组奶牛饲喂相同日粮,预试期15 d,正试期15 d。结果表明:泌乳盛期组奶牛每天干物质采食量(24.0 kg)高于泌乳后期组(P<0.01),泌乳盛期组奶牛每千克干物质的采食时间、一昼夜总反刍时间、躺卧时间均低于泌乳后期组(P<0.01),;泌乳盛期组奶牛日粮中中性洗涤纤维和酸性洗涤纤维的表观消化率低于泌乳后期组(P<0.01)。由此可见,泌乳盛期奶牛采食时间极显著高于泌乳后期奶牛,泌乳盛期奶牛躺卧时间极显著少于泌乳后期。     

不同分割液对奶牛不同阶段胚胎分割效果的影响

本试验探讨了3种不同分割液对奶牛桑葚胚和囊胚分割效果的影响。借助显微操作仪，将发育至第6～8天的体内常规生产的桑葚胚和囊胚进行分割，体外培养半胚，观察其发育情况，选择形态恢复好的半胚与一个囊胚滋养层细胞囊泡（trophoblastic vesicles，TRV）共移植。结果显示，在PBS+0.2 mol/L蔗糖与PBS+5%PVP中分割桑葚胚，其分割成功率显著高于PBS（P<0.05），分别为89.13%、86.73%和69.67%，而半胚的囊胚发育率及移植妊娠率三者均无显著差异（P>0.05）；在PBS+0.2 mol/L蔗糖与PBS+5%PVP中分割囊胚， 其分割成功率显著高于PBS（P<0.05），分别为94.52%、92.52%和70.52%，而半胚培养的囊胚发育率及移植妊娠率三者均无显著差异（P>0.05）；说明在PBS中分别添加0.2 mol/L的蔗糖和5%的PVP有利于提高奶牛桑葚胚和囊胚的分割成功率。     

抑制素被动免疫对黄体中期山羊生殖激素分泌的影响

为了探讨抑制素在山羊生殖激素分泌调节中的作用 ,本试验用抗抑制素血清对黄体中期雌山羊进行了被动免疫试验。将酪 30 -抑制素 α(1,30 ) NH2 与兔血清白蛋白螯合 ,用去势雄山羊制备抗抑制素血清 (INH- AS) ,最终滴度为 1∶96 0 0 0。对照血清来自牛血清白蛋白免疫的去势雄山羊。 12只黄体期日本 SHIBA山羊 ,肌注 PGF2α(2 mg/只 )进行同期发情处理 ,处理后 3～ 5 d母羊出现发情。在发情后 10 d(发情当天为 0 d) ,将母羊分为 2组 ,每组 6只。试验组颈静脉注射 INH- AS(2 0 m L /只 ) ,并在注射前 (0 h)和注射后 6、12、2 4、4 8、72、96、12 0 h采取外周血样 (肝素抗凝 ) ;对照组注射对照血清 (2 0 m L/只 )。抗凝血样离心 (4℃、170× g、15 min)分离血清 ,- 2 0℃保存待测。血清 FSH和 L H含量用NIDDK放免试剂盒测定 ,其中 FSH标记纯品为 NIDDK- o FSH- I- 1,标准品为 NIDDK- o FSH- RO- 1,一抗为 NIDDK-抗- o FSH- 1;L H标记纯品为 NIDDK- o L H- I- 3,标准品为 NIDDK- o L H- RP- 2 4 ,一抗为抗 - o L H- YM。二抗为山羊抗家兔Ig G。血清 E2 和 P含量用1 2 5I放免盒测定。测 E2 时 ,为除去游离脂肪 ,样品用 0 .5 m L乙腈 +2 m L乙烷处理。试验结果表明 ,注射 INH- AS后 12 h,血清 FSH含量显著提高     

Effects of Breed on Follicular Dynamics and Oestradiol Secretion during the Follicular Phase in Sheep

Current study evaluates effects from breed and management background on follicular dynamics and endocrine output during the follicular phase of sheep oestrous cycle. Follicular phases were synchronized with three cloprostenol doses, 10 days apart, in three groups of 10 females of different non-prolific Spanish breeds (Manchega, Rubia del Molar and Negra de Colmenar). Development of all follicles reaching antral diameters >or=2 mm was assessed by daily transrectal ultrasonographies, whereas follicular function was evaluated by measurement of plasma oestradiol concentrations. All the ovulatory follicles were present at induced luteolysis in Manchega sheep, while a 93.7% were detected in Rubia del Molar and Negra de Colmenar ewes. The mean size of these ovulatory follicles was similar between breeds at 0 h, but their growth rates were higher in Manchega ewes, reaching a larger size at oestrous detection than in Negra de Colmenar and Rubia del Molar sheep (p < 0.05). Conversely, the oestradiol levels increased with time in Rubia del Molar and Negra de Colmenar (p < 0.05); whilst remained stable in Manchega females. However, the patterns of follicular turnover were similar between breeds. These results indicate that, though differences in follicular size and size distribution, patterns of follicular turnover in sheep are affected neither by the breed nor by the background of management and selection.     

Milk Progesterone Profile at and after Artificial Insemination in Repeat-Breeding Cows: Effects on Conception Rate and Embryonic Death

The aim of this study was to investigate whether the skim milk progesterone concentrations at artificial insemination (AI) and day of rise of post-ovulatory progesterone concentration thereafter affect the conception and embryonic death rates in repeat-breeding cows. Milk samples were obtained from 96 repeat-breeding cows that failed to conceive to three or more AIs. The samples were taken from the cows at the day of AI and three times/week until day 45 post-AI. Skim milk was obtained after centrifugation and used for progesterone assay. The cows with a progesterone concentration more than 0.5 ng/ml at AI showed a significantly higher incidence of late embryonic death than those having a progesterone concentration<0.5 ng/ml at AI (p<0.01). As the progesterone level at insemination rose, conception rate declined. A negative correlation was shown between conception rate and skim milk progesterone level at AI. Of 56 cows showing a rise of progesterone to 1 ng/ml or more within 6 days after AI, 28 cows (50%) conceived. On the contrary, only eight of 39 cows (20.5%) conceived when the progesterone rose up to 1 ng/ml after day 6 post-AI. We concluded that increased progesterone concentration at the time of AI and delayed rise of progesterone post-AI might lead to decrease in fertility in repeat-breeding cows.     

奶牛奶中孕铜含量变化模式及其对繁殖率的影响

通过３３２头奶牛孕酮含量变化周期的分析,获得孕酮谷点超前、居中和滞后３种模式,其排卵对应有超前、适中和延缓３种。这３种模式的奶牛情期受胎率分别为１９．２％,６３．６％和４１．４％。奶牛配种后,怀孕和未孕对孕酮含量变化有影响：怀孕牛,配种后从孕酮谷点至第１４天孕酮含量有提高（Ｐ＜０．０５）,特别是第１４天提高更显著（Ｐ＜０．０１）;未孕牛,配种后除谷点处孕酮含量较一致外,其余各处平均值都下降（Ｐ＜０．０５）。     

荷斯坦牛不同泌乳阶段累计产奶量和305d产奶量的相关性分析

为了解荷斯坦牛不同泌乳阶段累计产奶量和305 d产奶量的相关性,利用单因素方差分析法对806头荷斯坦牛不同胎次各泌乳阶段累计产奶量和305 d产奶量进行分析.结果表明:胎次对不同泌乳阶段累计产奶量和305 d产奶量均有极显著影响(P<0.01),5胎305 d产奶量最高,1胎最低,且极显著低于其它胎次(P<0.01);...     

添加不同水平苜蓿对奶牛生产性能及经济效益的影响

选取河北省沧州地区奶牛养殖小区体重、产奶量、胎次、泌乳期相近的荷斯坦奶牛40头,随机分为4组,对照组日粮组成为秸秆加精料,试验组添加不同水平苜蓿干草,分析对奶牛生产性能及经济效益的影响.结果表明,试验组与对照组相比产奶量显著提高,差异极显著(P<0.01);乳脂率降低,差异不显著(P>0.05);乳蛋白、非脂乳固形物极显著提高(P<0.01);体细胞数显著下降(P<0.05).添加苜蓿使牛奶品质得到明显改善.经济效益分析表明,日粮中添加苜蓿能显著提高奶产业种、养、加链条的整体效益.