Fecal progestagens to detect and monitor pregnancy in captive female cheetahs (Acinonyx jubatus)

The purposes of the present study were to establish a noninvasive monitoring assay of fecal progestagen measurement to detect pregnancy and to identify the components of fecal progestagens in early, middle and late pregnancy in cheetahs. Feces were collected from 7 female cheetahs and analyzed from 30 days before the last copulation to parturition in 9 pregnancies. Blood was collected from one cheetah. Fecal progestagen and serum progesterone concentrations were determined by enzyme immunoassay (EIA). The profiles of the fecal progestagen concentrations were similar to the serum progesterone profile. Fecal progestagen and serum progesterone concentrations remained at the baseline until copulation. In the mean fecal progestagen profile during pregnancy (92.8 ± 0.4 days; from the last copulation to parturition), the concentrations increased 3-4 days after the last copulation and remained high until parturition. To investigate changes in the components of progestagen metabolites in the tripartite periods of gestation, fecal progestagens were analyzed by HPLC-EIA. Marked immunoreactive peaks consistent with 5α-pregnan-3α/β-ol-20-one and 5α-pregnan-3,20-dione and small peaks consistent with 5β-pregnan-3α/β-ol-20-one were detected. There were no distinct difference in the components of progestagens among the first, second and third trimesters of pregnancy. The hormone assay, as an indicator of fecal 5α-reduced pregnanes, is useful for detecting pregnancy and monitoring pregnant luteal activity in cheetahs.     

Noninvasive reproductive monitoring in the okapi (Okapia johnstoni).

Fecal progestagen analysis in okapis (Okapia johnstoni) was used for diagnosis of pregnancy and reproductive disorders, including a comparison of urinary and fecal progestagen analysis and endocrine data on the postpartum period. Data were generated on reliability of fecal progestagen analysis in early pregnancy diagnosis, and case reports were compiled involving single animals with missing luteal activity, abortion after twin pregnancy, and abortions due to deficient placental progestagen production. There was approximately 100-200-fold higher progestagen concentration in feces than in urine, thus explaining the high reliability of fecal progestagen evaluations in diagnosing luteal function and pregnancy. The postpartum period was characterized by lactational anestrus of several months duration, and a postpartum estrous cycle about 2-3 wk after parturition was observed in two of eight animals. An animal with five abortions due to deficient placental progestagen production was treated with altrenogest in a subsequent pregnancy and carried the fetus to term.     

Relationship between parity and mineral status in dairy cows during the periparturient period

Data from 65 cows were collected to clarify the relationship between parity and mineral status in Holstein cows during the periparturient period. Milk fever occurred in a second‐ and a fourth‐lactation cow at parturition. Bodyweight of cows decreased immediately after parturition, and the decreased weights of cows from 1 to 6 days after parturition ranged from 11 to 27 kg. Milk yield of multiparous cows was higher than that of primiparous cows, but Ca and P concentrations in colostrum were not affected by the parity. Plasma glucose of primiparous cows was higher than that of multiparous cows, and plasma nonesterified fatty acids of fourth‐and‐more‐lactation cows at parturition was highest. Plasma Ca concentration of cows at parturition decreased with advancing parity, and plasma inorganic P concentration of third‐and‐more‐lactation cows was lowest at parturition. Plasma parathyroid hormone of fourth‐and‐more‐lactation cows at parturition was highest, but plasma hydroxyproline and alkaline phosphatase was lowest. These results suggest that the large transfer of Ca and P to colostrum is a factor in the development of milk fever in third‐and‐more‐lactation cows.     

Pregnancy diagnosis in Swine: a comparison of two ultrasound instruments

This investigation was designed to assess the sensitivity, specificity and positive predictive values of amplitude-depth and Doppler pregnancy detectors when utilized for pregnancy diagnosis of mated sows. Mated sows (n = 38) were tested daily from 15 to 45 days after mating for pregnancy with the ultrasound instruments. The same procedure was performed with nonmated sows (n = 10) 15 to 45 days after estrus. Both instruments were unreliable for pregnancy diagnosis between 15 and 22 days after mating. Between 23 and 45 days after mating the Doppler pregnancy detector was more specific and had greater positive predictive values. In contrast the amplitude-depth pregnancy detector was more sensitive during the same time interval.

Serum estrone sulphate concentrations were determined in samples collected between 27 and 30 days after mating or estrus, as an alternate method of pregnancy diagnosis. Serum estrone sulphate concentrations were always equal to or greater than 0.5 ng/mL in the pregnant sows, while in the nonmated sows estrone sulphate concentrations were never more than 0.5 ng/mL serum.

     

Excretion patterns of fecal progestagens, androgen and estrogens during pregnancy, parturition and postpartum in okapi (Okapia johnstoni)

The aim of the present study was to establish a simple method to monitor ovarian activity and non-invasively diagnose pregnancy in okapi (Okapia johnstoni). The feces of a female okapi were collected daily or every 3 days for 28 months. Steroids in lyophilized feces were extracted with 80% methanol, and the fecal levels of immunoreactive progestagens (progesterone and pregnanediol-glucuronide), androgen (testosterone), and estrogens (estradiol-17beta and estrone) were determined by enzyme immunoassays with commercially available antisera. Using the progesterone profiles, the durations of the luteal phase, follicular phase, and estrous cycle were determined to be 11.1 +/- 0.4, 5.3 +/- 0.6, and 16.5 +/- 0.7 days (n=22), respectively. Fecal levels of immunoreactive progesterone, pregnanediol glucuronide, and testosterone gradually increased from early pregnancy and peaked several months before parturition. More pregnanediol glucuronide was excreted in feces than progesterone during late pregnancy, but not during the estrous cycle. Although the fecal concentrations of immunoreactive estradiol-17beta and estrone change a little throughout pregnancy and non-pregnancy, they rose sharply and temporarily on the day following parturition. The present study indicates that fecal assays with commercial antisera for progesterone and pregnanediol glucuronide are useful for evaluating luteal activity and diagnosing pregnancy and indicates that estrogens might have some role as a trigger of parturition.     

Fecal estrone sulfate profile in sows during gestation.

The aim of this study was to establish radioimmunoassay (RIA) for fecal estrone sulfate (E1S) and to elucidate changes in fecal E1S during pregnancy in the sow. Fecal E1S was extracted on a commercially available solid phase column, and the E1S fraction obtained was subjected to RIA. The sensitivity of the RIA was 8.5 pg/tube. The intra- and inter-assay coefficients of variation were 8.8-8.9% and 10.7-14.2%, respectively. Mean recovery for E1S added to fecal samples was as high as 95.0%. A significant positive correlation (r = 0.904, n = 147 p < 0.0001) existed between fecal and plasma E1S concentrations. Mean E1S concentration in feces and plasma fluctuated exhibiting two peaks. The first peak of E1S concentration was evident on day 28-32 in feces and on days 26-30 in plasma. The E1S concentration in both feces and plasma remained at baseline levels during mid-pregnancy, but began to rise gradually around days 72-82 and 70-80, in feces and plasma respectively, and reached a peak concentration on days 110-114. Following parturition, the concentration of E1S in plasma declined rapidly, but there was a two-day delay before a decline in fecal E1S. Apart from this two-day delay, changes in fecal E1S were similar to those in plasma E1S. The study indicates that the measurement of E1S in feces could be a useful tool for early pregnancy diagnosis and for monitoring fetal development in sows and gilts.     

Successful Treatment for Subinvolution of Placental Sites in the Bitch with Low Oral Doses of Progestagen

Subinvolution of placental sites (SIPS) is the major cause of persistent sanguineous vaginal discharge after parturition in the bitch. Spontaneous remission is common but may take several months, and hence, medical therapy to end the discharge is often requested. In this retrospective study, we evaluated the effect of treatment for SIPS with low oral doses of a progestagen. Nine bitches with SIPS, but otherwise clinically healthy, were found in the computer database of the Department of Clinical Sciences of Companion Animals. Seven of these bitches were treated with low oral doses of a progestagen (megestrol acetate, 0.1 mg/kg body weight (bw) once daily for the 1st week, then 0.05 mg/kg bw once daily for the 2nd week). The other two bitches were untreated. Treatment results were evaluated by a telephone questionnaire. Progestagen treatment was successful in all of the treated dogs; sanguineous vaginal discharge stopped within the treatment period. One of the two untreated dogs remained symptomatic until the next oestrus, approximately 120 days after parturition, and the other remained symptomatic until 6 weeks before the start of the next pro‐oestrus, 270 days after parturition. No side effects of the progestagen treatment were observed. Subsequent gestations, parturitions and puerperal periods of 5 mated bitches were uneventful. One bitch did not become pregnant after mating. In conclusion, the results of this study indicate that oral administration of low doses of progestagen for 2 weeks is effective in stopping persistent sanguineous vaginal discharge in bitches with SIPS, with neither side effects nor reduced subsequent fertility.     

圈养海南坡鹿的繁殖行为与粪样中孕酮含量的关系

利用放射免疫测定法测定了3只坡鹿粪便中孕酮含量的变化,观察记录了圈养坡鹿的繁殖行为。结果显示:圈养坡鹿繁殖行为具有明显的季节变化,如雄鹿的求偶行为、标记行为和雄鹿、雌鹿的性行为等在发情期表现明显。坡鹿怀孕及分娩前后,粪便中孕酮含量有明显的变化。怀孕前粪便中孕酮的含量维持在较低的浓度水平。怀孕后,孕酮含量明显上升。分娩后,孕酮含量迅速降至低浓度水平。粪便中孕酮含量的变化可以及时地反映坡鹿的生殖生理状况(包括发情、怀孕、分娩等)并作为早期妊娠诊断的依据。     

Changes in fecal progestagen profile after excretion in miniature pigs

The aim of this study was to evaluate whether the fecal progestagen (progesterone and its metabolites) levels of miniature pigs would change after excretion at room temperature. Our initial investigation focused on the correlations between the fecal progestagen concentrations with and without ether extraction and between the plasma progesterone and fecal progestagen concentrations in order to develop an enzyme-linked immunosorbent assay (ELISA) for fecal progestagen without ether extraction. There were significant correlations between fecal progestagen concentrations with and without ether extraction (r=0.880) and between fecal progestagen concentrations without ether extraction and plasma progesterone (r=0.763). The fecal progestagen concentration obtained by ELISA without ether extraction was almost identical to that obtained with ether extraction. These results validate the ELISA method without ether extraction, which was therefore used for the latter experiment. Fecal samples collected from the pigs were preserved for 0-24 h at room temperature, and then their fecal progestagen concentrations were measured. The fecal samples preserved for 0 to 24 h were analyzed by high performance liquid chromatography (HPLC) and ELISA. The concentrations of all samples significantly increased with time after preservation. The progestagen concentration of fresh feces (0 h) with high progestagen concentration (>1000 ng/g) increased significantly after 3 h. The concentration increased significantly after 12 h for fresh feces containing about 500 ng/g progestagen. HPLC analysis is showed that the fecal progesterone concentration, but not its other metabolites, doubled 24 h after excretion compared with the concentration at 0 h. These results suggest that dynamic changes in the profile of progesterone metabolites occur in feces after excretion.     

Observations on Ovarian Structure and Development of the Southern Giraffe,Giraffa Camelopardalis Giraffa

The occurrence of vesicular follicles and corpora lutea (CL), without evidence of ovulation, in late fetuses and during prepubertal life was confirmed in the southern giraffe. In this, and other respects, the pattern of ovarian development is similar in the southern giraffe to the northern subspecies. This early ovarian activity recedes during late prepubertal life. Numerous accessory CL were present in pregnant and non-pregnant giraffe but they did not attain the size of the true CL of pregnancy. The latter undergo an initial phase of rapid enlargement lasting about 100 days, followed by a period of recession till mid-pregnancy and a second growth phase which persists until term (457 days). The CL of pregnancy rapidly regresses after parturition. The accessory CL regress slowly and form normal looking corpora albicantia (CA). These CA are most abundant during early adult life as also are vesicular follicles.     

Plasma steroid hormone concentrations and their relationships in Suffolk ewes during gestation and parturition

In this study, we measured plasma concentrations of progesterone, pregnenolone, estradiol, estrone, estrone sulfate, and cortisol and analyzed the correlations between these hormones during gestation in 13 Suffolk ewes, the main breed in Japan. Progesterone increased during gestation and decreased a few days before parturition; however, this pattern was different in samples with high progesterone concentrations (P4 spike samples). This P4 spike was associated with a high pregnenolone concentration. Apart from the P4 spike, the progesterone change was similar to that in other sheep breeds. Pregnenolone increased during gestation and decreased after parturition. A significant correlation between progesterone and pregnenolone was observed a few days before parturition. Estrone sulfate and estradiol concentrations increased during gestation, but estrone did not. They increased shortly before parturition, and then decreased immediately after parturition. At parturition, the correlation between estrone and estrone sulfate was significantly stronger. Moreover, a strong correlation between estrone sulfate and estradiol was observed after parturition. Cortisol did not change during gestation and increased shortly before parturition. The results showed steroid hormone dynamics in normal pregnant Suffolk ewes, which were mostly in line with those of other sheep breeds. It should be noted that high progesterone concentrations altered the typical patterns.     

Prostaglandin Release at Dexamethasone Induced Parturitions in Cows

Three cows of the Swedish Red and White Breed (SRB) were used in the experiment. Cow no. 1 pregnant for 247 days, was given 20 mg dexamethasone twice with an interval of 48 hrs. Cows nos. 2 and 3, each pregnant for 254 days, received 20 mg of dexamethasone twice with an interval of 24 hrs. The cows delivered normal living calves 153, 138 and 137 hrs., respectively, after the second injection of dexamethasone. Blood samples were drawn from the jugular vein every third hour during the experimental period and the samples were analyzed for estrone, progesterone and 15-keto-13,14-dihydroprostaglandin F₂α. Following the dexamethasone injections there was a continuous increase in the blood plasma levels of estrone followed by a sharp decrease in conjunction with parturition. The blood plasma level of progesterone showed a slow but continuous decrease until about 24 hrs. before delivery when a marked drop occurred. The levels of the prostaglandin metabolite increased gradually until about 24 hrs. prior to delivery. This was followed by an abrupt rise, and high levels of the prostaglandin metabolite were recorded for up to four days following parturition. It is concluded that the estrone increase preceded that of the prostaglandin metabolite and that the final drop in the progesterone was synchronous with the final rise of the prostaglandin metabolite level.     

The effect of dietary protein on reproduction in the mare. VI. Serum progestagen concentrations during pregnancy

Sixty-four Thoroughbred and Anglo-Arab mares aged 6-12 years were used, of which 40 were non-lactating and 24 lactating. Foals from these 24 mares were weaned at the age of 6 months. Non-lactating and lactating mares were divided into 4 dietary groups each. The total daily protein intake and the protein quality (essential amino-acid content) differed in the 4 groups of non-lactating and 4 groups of lactating mares. The mares were covered and the effect of the quantity and quality of dietary protein on serum progestagen concentrations during pregnancy was studied. A sharp decline in serum progestagen concentrations was recorded in all dietary groups from Days 18 to 40 of pregnancy, with some individual mares reaching values of less than 4 ng/ml. Serum progestagen concentrations recorded in some of the non-lactating mares on the low-quality protein diet increased to higher values (p < 0.05) than those of mares in the other 3 dietary groups at 35-140 days of pregnancy. A similar trend was observed for the lactating mares on a low-quality protein diet at 30-84 days of pregnancy. No such trends were observed in any of the other dietary groups. High-quality protein supplementation increased serum progestagen concentrations during the 1st 30 days of pregnancy. Lactation depressed serum progestagen concentrations until after the foals were weaned.     

前列腺素诱导母猪分娩时的内分泌状态

选用５头妊娠后期母猪,于妊娠１１１ｄ上午９∶００肌肉注射氯前列烯醇（ＰＧ）０．２０ｍｇ。５头母猪皆于次日白天分娩。从注射前至分娩结束,在整个诱导过程中多次采血,监测了血清中孕酮（Ｐ４）、硫酸雌酮（Ｅ１Ｓ）、催乳素（ＰＲＬ）和皮质醇（ｃｏｒｔｉｓｏｌ）４种激素浓度的变化。结果表明,诱导分娩过程中各种激素的变化和自然分娩基本一样。这就解释了用国产前列腺素诱导母猪分娩,只要处理时间合适（妊娠１１１ｄ或１１２ｄ）,就会有良好效果,不会产生不良副作用。     

Plasma Levels of Progesterone and Oestradiol-17 β in Reindeer (Rangifer Tarandus Tarandus) During Pregnancy

The plasma concentrations of progesterone and oestradiol-17p during pregnancy and the first 20 days after parturition were estimated in reindeer. The concentration of progesterone in the period 75–25 days prior to parturition was significantly higher than in the period 200– 75 days prior to parturition (P < 0.001). During the last 25 days before parturition the concentration decreased significantly. The concentration of oestradiol-17β was in most cases below 70 pg/ml until 50 days prior to parturition. During the last 25 days of pregnancy there was a significant increase in oestradiol-17β and the ratio progesterone/ oestradiol-17β was markedly lower than in the period 75–25 days before calving.Key words: reindeer, gestation, plasma, progesterone, oestradiol-17β     

Risk of abortion associated with Neospora caninum during different lactations and evidence of congenital transmission in dairy cows

OBJECTIVE: To examine the relationship between exposure to Neospora caninum and abortion in dairy cows during their first, second, third, and fourth or later lactations and to establish the main mode of transmission in female calves from birth until their first pregnancy was terminated by abortion or parturition. DESIGN: Prospective observational study. ANIMALS: 460 Holstein cows and 79 female calves. PROCEDURE: Cows were classified as seropositive or seronegative to N caninum within 7 days after calving; incidence of abortion was compared between groups during different lactations. Blood samples were collected from female calves before ingestion of colostrum and every 6 months until their first pregnancy was terminated by abortion or parturition; number of seropositive calves was compared between seropositive and seronegative dams. RESULTS: During the first pregnancy of their second lactation, risk of abortion for seropositive cows was 2.8 times that of seronegative cows. Among 10 calves born to seropositive cows, 4 were classified as seropositive at birth and thereafter. Among 69 calves born to seronegative cows, all were classified as seronegative at birth; 67 calves remained seronegative thereafter. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure to N caninum alone was not significantly associated with abortion in cows during the first, third, and fourth or later lactations. Seropositive cows that have aborted previously may have subsequent abortions attributable to N caninum. Congenital infection was the main mode of N caninum transmission in a cohort of female calves.     

Increase in plasma progestagen concentrations in the mare after foetal injection with CRH, ACTH or betamethasone in late gestation.

Maternal plasma progestagen concentrations increase about 20 days before parturition. The major contributors to the increase are reduced metabolites (ie 5 alpha-pregnanes). Precocious increases (ie less than 310 days of gestation) in these metabolites may occur in abnormal pregnancies. The effects of CRH, ACTH or betamethasone administered to the foetus at gestational ages ranging from about 250 to 320 days were examined. Sixteen healthy pony mares were used for foetal injection employing aseptic techniques. Water or normal saline were used as controls. Maternal plasma progestagen concentrations were measured using a commercial radioimmunoassay (RIA) progesterone kit and results were confirmed using gas chromatography-mass spectrometry (GC-MS). Results demonstrated clearly that an increase in maternal plasma progestagen concentrations occurred after injection of ACTH, CRH or betamethasone to the foetus, irrespective of gestational age. A comparable increase was not observed in the control animals. Of the 16 mares in which the foetus was injected, 13 produced viable foals at gestational ages ranging from 307 to 339 days whereas 3 mares delivered non-viable foals at 284 to 306 days gestation. The results support the hypothesis that the pre-parturient rise in progestagens occurring in the mare is the result of foetal adrenocortical activity.     

Measurement of urinary and plasma estrone sulphate concentrations from pregnant sows

The accuracy of urinary estrone sulphate (E1S) concentrations as an indication of pregnancy was compared to that of plasma E1S concentration. Urine and plasma were collected from 145 sows 18 to 30 days after mating. Concentrations of E1S in urine were indexed to creatinine concentration (CR) and specific gravity (SG) to correct for urine dilution. There was no significant difference in E1S concentration between urine corrected for CR or SG and uncorrected urine (UN) from pregnant sows. Urinary E1S concentrations were 20 to 100 times higher than plasma E1S concentrations. The E1S test was accurate in the detection of pregnancy in sows using both plasma and urine (test sensitivity, 98.8% vs. 96.4%, respectively) during the optimal sampling period of 20 to 30 days post mating. The test was slightly more accurate (NS) using plasma than urine in detecting non-pregnant sows (test specificity, 100.0% vs. 91.9%, respectively) 20-30 days after mating. Urinary E1S concentrations could be used to predict litter size, but the precision was poor ( +/- 3 piglets), and thus could only be used to predict small (less than 5 piglets), medium (6-10 piglets) and large (greater than 11 piglets) litters.     

围产期健康奶牛干物质摄入水平与血浆瘦素浓度及乳产量的关系

将健康的围产期奶牛 3 0头随机均分为 3组。其中Ⅱ组为对照组、Ⅰ组和Ⅲ组分别为试验组 ;Ⅰ ,Ⅱ ,Ⅲ 3组于产前 2 8d开始分别饲喂NRC标准减少 2 0 %日粮组 (能量摄入 80 %)、NRC标准日粮组 (能量摄入 1 0 0 %)、NRC标准增加 2 0 %日粮组 (能量摄入 1 2 0 %) ,产后各组奶牛均饲喂标准日粮 ,至产后 5 6d结束 ,观察不同干物质摄入水平对奶牛血浆瘦素浓度及产奶量、体重等生产性能的影响。试验结果表明 :奶牛产后干物质摄入量、血浆瘦素浓度和产奶量为Ⅰ组高于Ⅱ组、Ⅱ组又高于Ⅲ组 ,经统计分析组间差异极显著 (P <0 .0 1 )或差异显著 (P <0 .0 5 ) ;但产前2 8～ 1 4d瘦素浓度组间差异均不显著 (P >0 .0 5 ) ,此后Ⅰ组和Ⅲ组差异显著 (P <0 .0 5 )。此外 ,Ⅰ组体重消耗最小 ,Ⅲ组体重消耗最大 ,各组间差异极显著 (P <0 .0 1 )。可见 ,围产期健康奶牛不同干物质摄入量与血浆瘦素浓度、产奶量、体重等生产性能之间存在着密切的关系     

Direct enzyme immunoassay of fecal estrone derivatives in dairy cows

The present study was undertaken to develop a novel, practical and simple procedure for enzyme‐linked immunosorbent assay of fecal estrone derivatives (estrone, estrone sulfate and estrone glucuronide) in dairy cows. Fecal solution, prepared by mixing feces with borate buffer, was applied directly to wells without extraction, and incubated with anti‐estrone antibody and horseradish peroxidase‐labeled estrone. Estrone sulfate was used as the standard. The sensitivity of the assay was estimated as 0.15 ng/mL (0.6 ng/g). The intra‐assay and inter‐assay coefficients of variation were 5.3–8.1% and 13.4–15.7%, respectively. The recovery rate was 78–102%. Only 4 h were needed to complete an assay to measure fecal estrone derivative concentrations. A significant positive correlation was established between plasma estrone sulfate concentrations and fecal estrone sulfate equivalent concentrations. When fecal estrone sulfate equivalent concentrations were measured in pregnant dairy cows, a gradual increase from day 150 of pregnancy, and subsequent drastic increases from day 240 to calving date were observed. These results suggest that the direct enzyme immunoassay procedure developed in the present study is a practical and reliable method for measuring fecal estrone derivative concentrations.