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.     

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.     

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。     

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.     

添加外源性酶对猪、鸡内源消化酶活性的影响

观察了猪、鸡不同类型饲粮中添加外源性酶制剂后对其内源性消化酶活性的影响。结果,玉米－豆粕型饲粮中添加外源酶后,３７及６７日龄丝毛乌骨鸡的小肠胰蛋白酶活性分别提高１３．１５％（Ｐ＜０．０５）和９．４５％（Ｐ＜０．０５）,总蛋白水解酶活性提高７．３０％（Ｐ＞０．０５）和１３．７％（Ｐ＜０．０５）,但α－淀粉酶活性无显著变化;高麸皮饲粮中添加外源酶可使４９日龄ＡＡ肉鸡的胰蛋白酶,总蛋白水解酶和α－淀粉酶活性分别提高４６．５０％（Ｐ＜０．０５）,１５．２０％（Ｐ＞０．０５）和７９．８０％（Ｐ＜０．０５）,而对胰脏中α－淀粉酶、脂肪酶活性无显著影响;高次粉饲粮中添加外源酶可使杜长大仔猪的小肠总蛋白水解酶活性提高２１．００％（Ｐ＜０．０５）,而对肠α－淀粉酶活性无显著影响;大麦－豆粕型饲粮中添加外源酶可使杜长大肥育猪的肠胰蛋白酶活性下降３６．９０％（Ｐ＜０．０５）,肠脂肪酶活性提高７２．７％（Ｐ＜０．０５）,而对小肠α－淀粉酶和胰脏α－淀粉酶、脂肪酶活性无显著影响。据此认为,外源性酶与畜禽内源性消化酶活性的关系较为复杂,受到动物品种和饲粮类型的影响,尤其是后者。     

Effect of a Nonsurgical Embryo Transfer Procedure and/or Altrenogest Therapy on Endogenous Progesterone Concentration in Mares

Endogenous progesterone levels may decline after transcervical embryo transfer in some mares. Progestogen therapy is commonly used to support endogenous progesterone levels in embryo transfer recipient mares or those carrying their own pregnancy. The goal of this study was to determine the effects of the transcervical transfer procedure and/or altrenogest therapy on luteal function in mares. Mares were assigned to one of six treatment groups: group 1 (untreated control; n = 7 cycles), group 2 (sham transfer, no altrenogest; n = 8 cycles), group 3 (sham transfer plus altrenogest; n = 8 cycles), group 4 (pregnant, no altrenogest; n = 9 mares), group 5 (pregnant plus altrenogest; n = 9 mares), and group 6 (nonpregnant plus altrenogest; n = 10 cycles). Mares in groups 4-6 were bred and allowed an opportunity to carry their own pregnancy. Blood samples were collected for 22 days beginning on the day of ovulation. Sham embryo transfer (groups 2 and 3, combined) did not result in a decline in endogenous progesterone levels compared with control mares (group 6). However, sham embryo transfer did result in luteolysis and an abrupt decline in endogenous progesterone levels in one of the 16 (6.2%) sham-transferred mares. Altrenogest therapy in sham-transferred mares (group 3) was associated with lower endogenous progesterone levels on days 10, 12, and 13 postovulation when compared with sham-transferred mares that did not receive altrenogest (group 2). Administration of altrenogest to pregnant mares (group 5) was associated with lower concentrations of endogenous progesterone from days 14 to 18 and on day 21 compared with endogenous progesterone levels in pregnant mares not administered altrenogest (group 4). In conclusion, a transcervical embryo transfer procedure can cause luteolysis in a low percentage of mares. Altrenogest therapy may be associated with a reduction in endogenous progesterone secretion, presumably mediated by a reduction in pituitary luteinizing hormone (LH) release and a decrease in luteotropic support.     

Luteal Characteristics and Progesterone Production on Day 5 of the Bovine Oestrous Cycle

In this study we have examined luteal function in non-lactating and late lactation dairy cows on day 5 of the cycle, during the period of the post-ovulatory progesterone rise. Comparison of luteal progesterone content and in vitro synthetic capacity with circulating plasma progesterone demonstrated that circulating progesterone concentration is a function of total luteal activity rather than the activity of individual units of tissue. Incubation of luteal tissue in vitro demonstrated stimulatory activity of LH and IGF-I, and to a greater degree IGF-II, on luteal progesterone synthesis. Finally the study showed no effect of double ovulation on luteal function. Occurrence of double ovulation in 35% of animals was not associated with any difference in luteal function or plasma progesterone concentrations.     

日粮中添加植物油对奶牛瘤胃代谢的影响

试验以3头年龄相同、平均体重为(650±20)kg、处于同一泌乳期且装有永久性瘤胃瘘管的荷斯坦奶牛为试验动物,采用3×3拉丁方设计,探讨日粮中添加不同的植物油(日粮脂肪含量均为6.5%)对奶牛瘤胃内纤毛虫数量、pH值、NH3-N浓度、菌体蛋白浓度及挥发性脂肪酸(VFA)浓度动态变化的影响。3种植物油分别为葵花油、胡麻油、菜籽油。试验结果表明:日粮中添加不同的植物油,瘤胃液纤毛虫数量动态变化的影响差异不显著(P>0.05),其平均值依次为(7.70,9.26,9.22)×105个/mL;对瘤胃液pH值动态变化的影响差异不显著(P>0.05),其平均值依次为6.16,6.21,6.15,均在瘤胃发酵的正常范围之内;瘤胃液NH3-N浓度动态变化3组之间差异不显著(P>0.05),其平均值依次为14.98,16.45,16.48 mg/100 mL;对菌体蛋白浓度的影响差异不显著(P>0.05),依次为85.17,89.83,78.87 mg/100mL。对VFA浓度的影响差异不显著(P>0.05)。     

Use of Radioimmunoassay to Measure Progesterone Levels During Different Reproductive Stages in Female Damascus Goats

Tropical Animal Health and Production -     

Different Types of Postpartum Luteal Activity Affected by the Exposure of Heat Stress and Subsequent Reproductive Performance in Holstein Lactating Cows

The aim of the present study was to determine the effect of heat stress on postpartum (PP) luteal activity and subsequent reproductive performance in lactating cows. Thirty Holstein–Friesian (HF, ≥75%) cows (hot season, n = 15 and cool season, n = 15) were studied over 60 days after calving. The changes of temperature‐humidity index (THI) were monitored within both seasons. Body condition scores (BCS), uterine involution and the ovarian structures were recorded. Plasma samples were obtained three times a week for the analysis of progesterone (P4) and prostaglandin F_2α metabolite (PG metabolite). Subsequent reproductive performance of the cows, calved within the same period was also evaluated. The means of THI for hot and cool seasons varied between 84–87 and 78–83 respectively. A drop of the mean BCS was recorded at 5 weeks PP in the cows during hot season (p < 0.01), however a delay of uterine involution during early PP period in hot season was not clearly seen. The cows with normal PP ovarian cyclicity during hot and cool seasons were 4/15 (26.7%) and 9/15 (60.0%) respectively. A higher percentage of abnormal luteal activity was found in the cows during hot season (p = 0.07) and delayed luteal cyclicity/anovulation was the most pronounced atypical P4 profile. The levels of PG metabolite were not different between groups and the relationship between the levels of PG metabolite and the time of uterine involution was not evident (p > 0.05). The heat detection rates and the pregnancy rates were higher in the cows during cool season (p < 0.05). Additionally, the higher first AI conception rates during cool season were recorded (p = 0.06). In conclusion, heat stress conditions had negative effects on BCS and altered a normal process of ovarian resumption PP, consequently resulted in lower reproductive performance in a tropical dairy herd.     

饲喂不同日粮对奶牛产奶量的影响研究

在规模养殖户饲养的奶牛中选择年龄在4～6岁,体重、胎次、泌乳时间等基本一致的荷斯坦泌乳牛18头,根据农户的饲喂条件,分为三组,每组6头。试验A组饲喂全价配合饲料和玉米秸秆青贮,试验B组饲喂混合饲料、粗饲料和玉米秸秆青贮,对照组C组采用一般的常规饲喂(有啥喂啥)方式,观察饲喂不同日粮对奶牛产奶量的影响效果。试验结果：试验A组奶牛头均日产奶量19．82kg,个体产奶量6045kg,试验B组头均日产奶量工17．27kg,个体产奶量5267kg,对照C组头均日产奶量12．41kg,个体产奶量3785kg;试验A组和B组头均日产奶量比对照C组增加7．41kg和4．86kg,个体产奶量分别增加2260kg和1482kg。