Effects of long-term consumption of sewage solids on reproductive performance and serum progesterone and estradiol-17beta in mature fine-wool ewes

Eighty-four mature fine-wool ewes were randomly allotted to one of three dietary treatments to examine the influence of dietary sewage solids on reproductive performance. Treatments were a basal diet or basal plus either 3.5% cottonseed meal (CSM) or 7% sewage solids. Serum progesterone and estradiol-17beta measured in blood samples collected through an estrous cycle just before the breeding season of the first and second years were not adversely influenced by sewage feeding. Progesterone concentrations quantified in weekly blood samples collected during gestation and lactation of both years did not differ (P>0.10) among treatments. During the first lambing season, lambing data showed that ewes consuming CSM produced slightly more lambs per ewe exposed (P<0.10) than either basal- or sewage-fed ewes. During the second breeding season, midventral laparotomies revealed that ovaries from ewes receiving CSM contained more (P<0.10) corpora lutea than did those from either basal- or sewage-fed animals. However, at the end of the second lambing season, no differences (P>0.10) among treatments in the number of lambs born or weaned were observed. A third year's lambing data also revealed no difference (P>0.10) among treatments for lambing rates. These data indicate that prolonged consumption of a diet containing 7% sewage solids has little, if any, detrimental effect on reproductive performance of mature fine-wool ewes.     

Concentrations of 13, 14-dihydro-15-keto-prostaglandin F(2)alpha, estradiol-17beta and progesterone during the peripubertal period in heifers

Twenty prepubertal Holstein heifers were utilized to assess plasma 13, 14-dihydro-15-keto-prostaglandin F(2)alpha (PGFM), serum progesterone (P(4)) and estradiol-17beta (E(2)) concentrations as well as the E(2):P(4) ratio during the onset of puberty in cattle. All animals were maintained as a group along with a sterile marker bull to assist in the detection of estrus. Upon detection of the first estrus (Day=O), daily blood samples were collected from a jugular vein until the heifers had completed 3 estrous cycles. The average body weight and age at first estrus were 247.6+/-4.8 kg and 304.0+/-7.5 days, respectively. Frequency of abnormal length estrous cycles was greater (P<0.02) during the first (40%) and second (35%) cycles than during the third estrous cycle (0%). All heifers had normal cycle lengths (18 to 24 days) by the third estrous cycle. Serum P(4) was greater during the third cycle (P<0.05) from Day 10 to Day 4 before the next estrus compared with the same period of the first estrous cycle. Serum E(2) did not peak until the day of estrus in the first cycle, whereas E(2) reached a maximal level 2 days before estrus in the third estrous cycle. Serum E(2) was higher (P<0.0001) 2 days before estrus in the third cycle than in the first estrous cycle. Plasma PGFM reached maximum concentrations 3 days before estrus in the third cycle compared with 1 day before estrus at the end of first estrous cycle. As estrus approached during the third cycle, PGFM rose 1 day before E(2) rose and P(4) declined, while the rise in PGFM and E(2) occurred simultaneously, with P(4) declining at the end of the first estrous cycle. During diestrus, the E(2):P(4) ratio was lower (P<0.07) in the third cycle than in the first, but it was higher (P<0.04) at estrus and 1 day before in the third estrous cycle. These data reveal a high incidence of abnormal length estrous cycles during the first two estrous cycles of the peripubertal period, and demonstrate anomalies in uterine and ovarian endocrine activity during the peripubertal period in cattle.     

Steroid levels after intramuscular injection of radioactive estradiol-17beta, estrone, progesterone and testosterone in the bovine

Eight 2 year old Hereford cows from days 8 to 12 of the estrous cycle were injected intramuscularly with 5 ml of corn oil containing 5 mg of estradiol-17beta (two cows), estrone (two cows), progesterone (two cows) or testosterone (two cows). Each cow treated with estradiol received 494 microc of estradiol-17beta-6, 7 H3 and each cow treated with estrone received 492 microc of estrone-6, 7 H3. Each cow treated with progesterone or testosterone received 400 muc of H3 compound labeled in the 7 position. Total urine was collected by urethral catheterization of the cows treated with estrogens. Blood samples for plasma and serum were collected via jugular cannulae. Blood and urine samples from estrogen-treated cows were collected hourly for the first 24 hr, at 2 hr intervals for the next 26 hr, at 4 hr intervals for the next 12 hr and at 12 hr intervals until background was reached. Blood samples were collected hourly from 1 to 8 hr after injection from progesterone or testosterone-treated cows. Plasma and serum levels of radioactive estradiol-17beta, estrone, progesterone and testosterone were similar. Blood levels of radioactivity peaked at 2 hr post-injection in cows receiving estradiol-17beta and at 3 hr in cows receiving estrone. Blood levels of labeled estradiol-17beta and estrone were nondetectable by 54 hr and 83 hr, respectively. Peak urinary excretion of radioactivity was reached at 7 hr for estradiol-17beta and at 14 hr for estrone and nondetectable levels were reached by 95 hr for estradiol-17beta and 14 hr for estrone. At these times, 15.5% of the total dose of radioactive estradiol-17beta and 17.5% of the injected estrone had been excreted in the urine. Peak blood and urinary excretion levels were reached earlier for radioactive estradiol-17beta than for estrone, and excretion of estradiol-17beta was completed more rapidly. No difference was found in plasma and serum levels for any steroid studies; thus, endogenous steroid titers in blood plasma and serum are not different in the cow.     

The relationship between plasma levels of estradiol-17beta, progesterone and the consistency of the cervix in the sow

The object of this investigation was to estimate the peripheral plasma levels of estradiol-17beta and progesterone and the consistency of the cervix by rectal palpation of sows before, during and after post-weaning estrus. The tonicity of the cervix increased from approximately 4 days before the onset of heat, became firmest during estrus, and became softer again during the post-estrual period. The levels of estradiol increased before the onset of heat and reached maximum levels on the day preceding heat, then decreased to baseline. The progesterone levels remained low until post-estrus, then increased rapidly. One sow which did not come in heat constantly had a soft cervix and low hormone levels. It is concluded that estrogen might be implicated in the increased tonicity of the cervix and that rectal examination of the cervix provides valuable information on ovarian function.     

The influence of estradiol-17beta and progesterone on peripheral serum concentrations of luteinizing hormone and follicle stimulating hormone in the ovariectomized ewe

Serum gonadotropin concentrations were high and variable and fluctuated episodically in short and long term ovariectomized ewes. Treatment with solid silastic implants releasing progesterone (serum levels 1.81 +/- 0.16 ng/ml) had no consistent effect. Treatment with implants releasing estradiol-17beta significantly depressed mean serum gonadotropin concentrations and peak height to values usually seen in intact ewes. This occurred regardless of implant size and serum estradiol-17beta concentrations (range 11 +/- 0.3 pg/ml to 98 +/- 12.8 pg/ml). Progesterone and estradiol-17beta together significantly depressed the frequency of peaks in LH concentration. Following progesterone removal, 95% of the ewes treated with progesterone and estradiol-17beta implants experienced a transient increase in serum LH concentrations similar to the preovulatory surge in intact ewes. Eighty-four percent of the LH surges were accompanied by a surge in serum FSH concentrations. However, following progesterone removal, 5.1 +/- 2.1 FSH surges were observed over six days. Gonadotropin surges occurred regardless of estradiol-17beta implant size and with or without the influence of supplemental estradiol-17beta.     

Repeated induction of abortion in bitches and the effect on plasma concentrations of relaxin, progesterone and estradiol-17beta

The aim of the present study was to investigate the effects of two medications on two subsequent abortions and plasma hormone concentrations of dogs. For this purpose, two groups of bitches (n=5 each), received the antiprogesterone aglepristone (Alizine) at 10mg/kg body weight on two subsequent days around day 30 after mating. In group II, the antiprolactin cabergoline (Galastop) was additionally administered po at 5 microg/kg body weight until the start of abortion. The plasma concentrations of relaxin, progesterone (P4) and estradiol-17beta (E2) were measured before, during and after each abortion. During the next cycle after the abortion, the same bitches were mated again and in pregnant animals, induction of abortion was performed as before. During the third cycle, pregnant bitches were allowed to whelp. Termination of first pregnancy occurred significantly earlier after the combined treatment (6.8 versus 10.6 days, p<0.05). In both groups and during both abortions, relaxin varied between individuals; however, there was a continuous decrease after the abortions and no significant differences between groups (p>0.05). In one bitch with high relaxin concentrations before treatment (11.6 ng/ml), a cystic endometrial hyperplasia was diagnosed. In the aglepristone only group, P4 concentrations increased significantly after the first application (p<0.05), then decreased continuously until day 45 after the beginning of abortion. In the combined group, there was a continuous decrease until day 45 (p>0.05). At this time, P4 concentrations between 0.47 and 84.9 nmol/l were measured in both groups. The level of E2 over time was not influenced by any medication. We therefore note that the two medications mainly influenced plasma concentrations of P4 in different ways, probably due to specific treatment-hormone interactions. However, all measurements fell within the range considered normal.     

Effect of holding time and temperature of bovine whole blood on concentration of progesterone, estradiol-17beta and estrone in plasma and serum samples

Bovine jugular venous blood was collected, with and without heparin, and aliquoted into 140 12-ml tubes. Four subsamples (two heparinized and two coagulated) were centrifuged immediately (time zero) and plasma or serum was aspirated and stored at -20 degrees C. One-half of the remaining subsamples were stored at 4 degrees C and the other one-half at 25 degrees C (room temperature). At 1-h intervals (0 to 24 h), 6-h intervals (24 to 72 h) and at 96 and 120 h, four subsamples (heparinized and coagulated at both 4 degrees C and 25 degrees C) were centrifuged, plasma or serum was aspirated and stored at -20 degrees C. Whole blood incubation for 1 h at 25 degrees C reduced mean plasma and serum progesterone (P(4)) concentration (P<0.05). Similarly, whole blood incubation at 4 degrees C for 2 and 3 h, respectively, reduced mean plasma and serum P(4) concentration (P<0.05). No difference was found in mean P(4) concentration between plasma and serum samples harvested from whole blood incubated at 4 degrees C or 25 degrees C. Concentration of estradiol-17beta (E(2)) and estrone (E(1)) fluctuated over time, irrespective of holding temperature. There was a blood type, heparinized or coagulated, by time interaction (P<0.01) for both E(2) and E(1) concentrations It was concluded that incubation time and temperature between collection and centrifugation of bovine blood samples influenced the assayable P(4) concentration in both plasma and serum. In contrast, incubation temperature had no effect on assayable E(2) and E(1) concentrations, but assayable E(2) and E(1) over time were differentially affected, depending on whether plasma or serum was assayed.     

The effect of active immunization against progesterone on plasma concentrations of total and free progesterone, estradiol-17beta and LH in the cyclic ewe

Nine mature cyclic ewes were actively immunized against progesterone which was rendered immunogenic by conjugation to bovine serum albumin (BSA). Seven control ewes were immunized with BSA. In ewes immunized against progesterone, the concentration of total plasma progesterone increased to 24.3 ng/ml vs 2.8 ng/ml in control animals (P<0.001). However, immunization did not affect the plasma levels of free, unbound progesterone. The correlation coefficient between total plasma progesterone concentrations on Days 4 to 11 of the estrous cycle and antibody titer was r=0.983. Estradiol-17beta concentrations in immunized ewes were higher than in controls on Days 6 to 15 of the estrous cycle (P approximately 0.05). Frequent sampling for LH over a 6-h period on Days 2, 5, 8, 11 and 14 of the cycle revealed no significant differences in the frequency and amplitude of LH pulses between immunized and control ewes. The immunized ewes had estrous cycles of normal length and maintained normal pregnancies. It is suggested that the immunized cyclic ewe is capable of maintaining adequate levels of free progesterone by greatly increasing progesterone synthesis, thus neutralizing the effect of the antibodies.     

Long-term effects of a single cadmium chloride injection on the ovulation, ovarian progesterone and estradiol-17 beta secretion in rats

On the day of dioestrus II rats were given 5, 10 or 15 mg/kg of cadmium chloride (CdCl2), or 1, 0 ml/kg of 0.9% NaCl solution. Then ovarian cycle was checked daily for a period of 12 cycle length. On the day of oestrus or expected oestrus in the 13th cycle the animals were anaesthetized with pentobarbital and cannulas were inserted in one of the femoral arteries and veins and in one of the utero-ovarian veins. Five-minute blood fractions were collected for 40 minutes and following the first blood samples 10 IU of hCG were injected iv. Ovarian venous outflow and blood pressure were continuously recorded. From the blood fractions progesterone (P) and oestradiol-17 beta (E2) were determined with RIA and the P and E2 secretion rates of ovary were calculated. Ovaries were excised and oviducts were flushed for counting oocytes. CdCl2 shortly after its administration induced a (dose-dependent) anoestrous period which turned into regular or irregular cycles depending on the dose. Part (28-32%) of the oestrous animals (14% that of the controls) remained unovulatory, when ovulation occurred normal number of ova was found. None of the doses of CdCl2 has influenced the blood pressure of animals and blood flow of the ovary. The basal secretion rate of P and E2 was not changed in the ovary compared to the controls. The hCG induced rise of P secretion, however, in the animals treated with 5 and 10 mg/kg bw CdCl2 was diminished and delayed, while in the animals treated with the 15 mg/kg Cd dose a complete lack of response was observed.     

Effects of corpus luteum removal on progesterone, estradiol-17 beta and LH in early pregnancy of the tammar wallaby, Macropus eugenii

The quiescent corpus luteum of female tammars was reactivated by removal of the pouch young (RPY). The reactivated corpus luteum was ablated 3 days after RPY. Plasma progesterone and oestradiol concentrations were measured by radioimmunoassay in these and in sham-operated controls. Excision of the CL abolished the rise in progesterone seen at Day 5-6 in the sham-operated animals (130.7 +/- 56.6 vs 452.4 +/- 176.0 pg/ml, mean +/- s.d.). By contrast, oestradiol-17 beta values increased within 6-16 h of CL excision to 16.3 +/- 6.9 pg/ml and remained high for 1-3 days while in the sham-operated animals there were less sustained and more variable peaks of 10-20 pg/ml between Days 3 and 5 (mean 12.0 +/- 3.6 pg/ml at Day 4-5). We conclude that the early transient increase in peripheral plasma of progesterone is of luteal origin but the source of the oestradiol remains unknown.     

The effect of cortisol on the synthesis of prostaglandins (PGF2 alpha, PGE2) by human endometrial fibroblasts in vitro with and without addition of estradiol-17 beta or progesterone

Cortisol is known as a potent inhibitor of phospholipase A2 activity in several tissues. In fibroblast monolayer cell cultures from proliferative human endometrium cortisol alone does not affect the basal PGF2 alpha or PGE2 synthesis. After stimulation of PGF2 alpha production by 10(-7) mol/l estradiol-17 beta increasing concentrations of cortisol up to 10(-5) mol/l dosedependently reduce the PGF2 alpha production. Also the progesterone (10(-4) mol/l) stimulated increase of PGF2 alpha and PGE2 synthesis is inhibited by cortisol (10(-7) mol/l).     

Action of estradiol-17 beta, estrone sulfate and progesterone on the growth and differentiation of endometrial epithelial cells of the guinea pig in primary culture

Endometrial guinea-pig glandular epithelial cells grown in primary culture incorporated [3H] thymidine. After three washings with a steroid-free medium, they were made quiescent and arrested in the G0/G1 phase. However, they remained hormone-responsive and resumed the cellular cycle after stimulation by 10(-6) M oestrone sulphate but not by oestradiol-17 beta.     

Variations in hepatic estradiol-17beta receptor concentrations during the annual reproductive cycle of diploid and triploid female catfish, Heteropneustes fossilis (Bloch)

Concentrations of hepatic estradiol-17beta (E2) receptors (ER) in cytosolic and nuclear fractions were evaluated in diploid and triploid female catfish Heteropneustes fossilis (Bloch) during four different reproductive periods of a complete reproductive cycle. Basal level of ER concentration was noted in the resting period of both diploids and triploids. Receptor level gradually elevated through the preparatory period and reached a peak in the pre-spawning period in both diploids and triploids. However, ER concentrations were overall reduced in triploid to that of diploid females. In a single point assay, in diploids, ER concentration showed about a 3-fold rise (p<0.001) in the cytosolic and a 4-fold rise (p<0.001) in the nuclear extracts from resting to the pre-spawning period. In triploids, only a 2-fold rise was observed both in cytosolic (p<0.01) and nuclear (p<0.05) ER concentration during the same span. Finally, a sudden fall of receptor level was observed in the spawning period in both the ploidy groups with a lower concentration in the triploids. The K(d) value did not differ between the females of diploids (cytosolic 1.12+/-0.21 nM and nuclear 6.9+/-0.9 nM) and triploids (cytosolic--1.13+/-0.17 nM, nuclear--6.8+/-2 nM). However, B(max) of the diploid showed about double the value than triploid females both in the cytosolic (diploid--367.4+/-33.24 pmol/mg protein, triploid--187.3+/-13.20 pmol/mg protein, p<0.001) and nuclear extracts (diploid--946+/-66 pmol/mg DNA, triploid-558+/-98 pmol/mg DNA, p<0.01) of liver. Lower E2 binding capacity and lower amount of E2 receptors of triploid catfish liver with a stunted vitellogenic status could be one of the major causes for reduced gonadal development and sterility in female triploids.     

Investigations on the effect of the ruminal N-balance on rumen metabolism, urea content in blood serum and milk as well as some liver parameters of lactating cows

Seven dairy cows fitted with ruminal and duodenal cannulae were used to investigate the influence of the amount of ruminally available N (Ruminal N-Balance, RNB) on the rumen metabolism and to answer the question on the lowest N-amount in the rumen, without negative effects on the fermentation. Animals were fed a ration on the basis of 7.9 kg corn silage and 7.2 kg concentrates related to dry matter, intended to meet the animals mean NEL and protein requirements. RNB amounted to -0.6 g/MJ ME in the basis ration. The other 3 rations were adjusted to RNB-values of -0.3, 0 and + 0.3 g/MJ ME by urea supplements in the concentrates. The increase in RNB resulted in higher NH3-N concentrations in the rumen fluid and in the duodenal digesta and higher urea concentrations in the blood and milk. The significantly highest amount of protein at the duodenum was detected when RNB showed an equilibrium (RNB = 0). The efficiency of microbial protein (MP) synthesis (gMP/kg fermented organic matter) was the same, g MP/d and g MP/MJ ME were significantly lower with RNB = -0.6g/MJ ME as compared to RNB = 0. The group with thelowest RNB showed the highest level of feedprotein degradation as well as the lowest organic matter, NDF and ADF fermentation. An effect on cholesterol, total bilirubin and gammaGT due to different RNB was not detected. The activities of GLDH and AST were highest when the RNB was -0.6 g/MJ ME. From the results, it can be concluded that significantly negative effects on rumen fermentation occur when RNB-values are below -0.3 g/MJ ME. However, a positive RNB did not increase t he degradation and synthesis capacity of the rumen micro-organisms as compared to RNB = 0.     

Uptake, distribution and binding of vertebrate and invertebrate steroid hormones and time-dependence of ponasterone A binding in Calliphora vicina. Comparisons among cholesterol, corticosterone, cortisol, dexamethasone, 5 alpha-dihydrotestosterone, 1,25-dihydroxyvitamin D3, ecdysone, estradiol-17 beta, ponasterone A, progesterone, and testosterone.

The presence of specific binding sites for radiolabelled vertebrate-type and arthropod-type steroid hormones was investigated in several organs including salivary gland, and central nervous system of third instar Calliphora vicina larvae by thaw-mount autoradiography. Ponasterone A, a 20-hydroxyecdysone agonist and 20-hydroxyecdysone are the only steroids which bind to nuclear high affinity binding sites. These binding sites are DNA associated while nucleoli show no tracer binding. Ecdysone, an endogenous 20-hydroxyecdysone precursor, is taken up by target cells but no significant nuclear binding occurs. 1,25-Dihydroxyvitamin D3 concentrates in cytoplasm only and its uptake is highest compared to all other steroids. Progesterone and testosterone show weak accumulation in the cytoplasm, while for cholesterol, corticosterone, cortisol, dexamethasone, dihydrotestosterone and estradiol-17 beta, no noticeable uptake occurs. For ponasterone A, a clear time dependence of uptake and intracellular distribution is visible, suggesting the existence and involvement of specific ecdysteroid uptake and transport mechanisms. These results suggest the presence of binding sites for various mammalian steroids in insects. Whether vertebrate steroid hormones or metabolites of them play a role in insects or whether the uptake and binding is based on chemical similarities alone without biological significance remains to be further investigated.     

Concentrations of LH, oestradiol-17 beta and progesterone in the peripheral plasma of swamp buffalo cows (Bubalus bubalis) around the time of oestrus

Peripheral plasma concentrations of LH, oestradiol-17 beta and progesterone were measured in 13 mature swamp buffalo cows at 4-h intervals from 36-40 h before until 36-40 h after the onset of oestrus. Mean LH concentrations increased sharply to a peak of 35 ng/ml and returned to basal levels of 5 ng/ml within a 12-h period beginning soon after the onset of oestrus. Mean oestradiol-17 beta concentrations were within the range 9-13 pg/ml from 36-40 h before until 12-16 h after the onset of oestrus, and within the range 7-9 pg/ml thereafter. Progesterone concentrations remained around 0.1 ng/ml throughout the sampling period. There were no significant differences in hormone concentrations or changes between cows that conceived and those that did not conceive to artificial insemination 12-24 h after onset of oestrus.