Concentration of IGF-I and estradiol-17beta in the blood plasma of pregnant cattle

Blood samples were collected from pregnant cows, heifers and also from non pregnant cows serving as controls. The determination of insulin-like-growth-factor-I (IGF-I) and estradiol-17 beta (E2) were performed immunologically. In the non pregnant cows E2 remained unchanged. IGF-I decreased around parturition and increased again commencing the 6th week of lactation. Compared to nonpregnant animals E2 but not IGF-I was slightly elevated during the first 10 weeks of pregnancy. During the 10th up to 20th week of pregnancy the mean values of IGF-I were increased as well as the ones of E2. During the late pregnancy the values of IGF-I in heifers are obviously more elevated as in lactating pregnant or non-pregnant cows; analogous high values were determined in pregnant cows only during the dry period. Before parturition even a negative correlation exists between IGF-I and E2. It is concluded that IGF-I is predominantly regulated by other factors.     

Peripartum changes in plasma estrone sulfate and estradiol-17beta profiles associated with and without the retention of fetal membranes in holstein-friesian cattle

The aim of this study was to evaluate the changes in plasma concentrations of estrone sulfate (E(1)S) and estradiol-17beta (E(2)beta) during the peripartum period (from day 10 prepartum to day 1 postpartum) associated with and without retention of fetal membranes (RFM) in Holstein-Friesian cattle (n=42). Plasma samples were analyzed for E(1)S and E(2)beta by ELISA. All parturitions were spontaneous and normal. Of 38 cattle delivering singletons, 29 had no RFM (singleton-normal group) and nine had RFM for more than 12 h (singleton-RFM group). Four cows gave birth to twins, and each twin had its own fetal membrane (FM). Two twinning cows expelled both FMs normally within 12 h (twin-normal group). In the remaining 2 twinning cows (twin-RFM group), the FM was expelled normally for one twin (first), while the FM of the other (second) was retained. There were no significant differences in the E(1)S concentrations or their increments from the concentrations on the preceding day between the normal and RFM groups of singleton cows on any peripartum day. The mean plasma E(2)beta concentrations on each day from day 10 to day 3 prepartum were significantly lower (P<0.05) in the singleton-RFM group compared with the singleton-normal group; however, on days 2 and 1 prepartum, the increments in the E(2)beta concentrations from the concentrations on the preceding days were significantly higher (P<0.05) in the singleton-RFM group than in the singleton-normal group. Thus, the plasma E(1)S concentrations just before parturition may not be associated with RFM. In the cows with RFM, the lower plasma E(2)beta concentrations that were found prior to day 2 prepartum may have been associated with immature placentomes, and the rapid rise in plasma E(2) beta within 1 to 2 days prior to calving may have produced asynchrony of placental and/or fetal maturation in relation to calving, thus resulting in RFM.     

Development and validation of a sensitive enzyme immunoassay (EIA) for blood plasma cortisol in female cattle,buffaloes, and goats

A highly sensitive enzyme immunoassay (EIA) that used the second antibody coating technique and the cortisol-horseradish peroxidase conjugate as a label for determination of free and total cortisol in blood plasma of dairy animals (cows, buffaloes, and goats) was developed. For biological validation of the EIA, blood samples were collected from the animals at 48 and 24 h before and 0, 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, and 132 h after dexamethasone administration. The EIA was performed directly with 20 μL of fresh plasma (for free cortisol) and also with 20 μL of heat-treated plasma (for total cortisol) after 1:5 dilutions with PBS. Cortisol standards ranging from 0.39 to 200 pg/well/20 μL were used, and the sensitivity of the EIA procedure was found to be 0.39 pg/well/20 μL, which corresponded to 0.02 ng/mL. In comparison with RIA the EIA was at least 4 times more sensitive and required 5 times less cortisol antiserum. In female cattle, buffaloes, and goats, the total, free, and bound plasma cortisol before dexamethasone administration was significantly (P < 0.05) higher than the total, free, and bound cortisol after dexamethasone administration. It can be concluded from these studies that the direct, sensitive EIA validated for estimating the free and total cortisol concentrations was sufficiently reliable and quick for studying the dynamics of cortisol distribution in blood plasma of dairy animals.     

Relationship between testicular transferrin and plasma estradiol-17beta concentrations of dogs with azoospermia and dogs with sertoli cell tumors.

Testicular Transferrin (Tf) and peripheral plasma estradiol-17beta (E2) concentrations were measured in 3 dogs with azoospermia (AZ dogs), 3 dogs with Sertoli cell tumors (SC dogs), and 5 normal male Beagles. The mean Tf concentrations in the testes of the AZ dogs and the affected testes of the SC dogs, and the plasma E2 concentrations in both these groups of dogs were significantly higher than the values in normal dogs (P<0.05, 0.01 and 0.01, respectively). Therefore, excessive E2 secretion by hyperfunctioning Sertoli cells is thought to have caused the azoospermia in the 3 dogs.     

Relationship of prepartum plasma concentrations of estrone sulfate and estradiol-17β with the weight of the calf and placental parameters in Holstein–Friesian cows

The aim was to evaluate the relationship of prepartum plasma estrone sulfate (E₁S) and estradiol‐17β (E₂β) concentrations with the weight of the calf and the placental parameters. Holstein–Friesian cows (n = 33) inseminated artificially with Japanese Black beef bull semen at Hiroshima University Farm in Japan were used for the experiment. Blood samples were taken every day from day 270 of gestation until the day after calving. The plasma samples were analyzed for E₁S and E₂β by enzyme immunoassay. The calf birth weight was taken immediately after calving. Complete fetal membranes were collected from 19 cattle and the weights of the placental components and the number of cotyledons were recorded. All 33 cattle delivered singleton normal calves. The prepartum plasma E₁S concentration was found to correlate significantly (P < 0.01) with the calf birth weight (r = 0.83), total fetal membrane weight (r = 0.81), cotyledonary weight (r = 0.79) and inter‐cotyledonary membrane weight (r = 0.64), but it did not correlate significantly with the number of cotyledons, whereas prepartum plasma E₂β was not found to correlate significantly with the weight of either the calf or any of the placental components except the number of cotyledons. In conclusion, prepartum plasma E₁S, not plasma E₂β, was found to correlate significantly with the weight of the calf and the placental components.     

Dynamic changes in plasma concentrations of gonadotropins, inhibin, estradiol-17beta and progesterone in cows with ultrasound-guided follicular aspiration

To elucidate the effects of ultrasound-guided transvaginal follicular aspiration, plasma concentrations of FSH, LH, inhibin, estradiol-17beta and progesterone, and folliculogenesis were examined in Holstein cows. Four clinically healthy cows with regular estrous cycles were scanned by ultrasound per rectum once a week for 9 weeks before the commencement of follicular aspiration. All visible follicles were divided into 3 categories based on their sizes (2 < or = small < 5 mm; 5 < or = medium < 10 mm, large > or = 10 mm). The follicular aspiration was started at random during the estrous cycle and conducted under epidural anesthesia induced with 5 ml of 2% lidocaine once a week for 6 weeks. The average number of total visible follicles > or = 2 mm in diameter at 7 days after aspiration (21.7 +/- 7.4, n = 24) was similar to that before starting aspiration (26.7 +/- 10.5, n = 36). Plasma inhibin and estradiol-17beta declined and fell to a trough on 1.5 days and returned to pre-aspiration values by 5 days after aspiration. Plasma concentrations of FSH increased and reached peak levels between 1 and 1.5 days after aspirations. Plasma concentrations of LH also increased and reached peak levels between 0.5 and 1.5 days after aspirations. Both plasma FSH and LH had returned to pre-aspiration levels by 5 days after aspirations. Plasma concentrations of progesterone did not change with the follicular aspiration. These results demonstrate that follicular aspiration decreases plasma concentrations of inhibin and estradiol-17beta, which in turn leads to a rise in plasma concentrations of FSH and LH. It is suggested that marked increases in plasma concentrations of FSH and LH after the aspiration stimulate the development and maturation of a new cohort of follicles within one week in cows.     

Validating a commercially available enzyme immunoassay for the determination of 17beta-estradiol and progestogens in the feces of cheetahs (Acinonyx jubatus): a case report.

Fecal 17beta-estradiol and progestogens excretion was monitored in adult, female cheetahs (Acinonyx jubatus; n = 2), ZGG-12301 (born 3 April 1993), gonadotrophin treated and ZGT-3301, (born 19 August 1993), nontreated, for 120 days using commercially available plate enzyme immunoassay kits prepared for human serum or plasma. There were significant differences (P < 0.001) between baseline and peak concentrations of both hormone measures. Female ZGG-12301, which conceived, but this pregnancy resulted in an unobserved spontaneous abortion, showed no significant difference (P > 0.05) between baseline and gestation 17beta-estradiol values; fecal 17beta-estradiol excretion during pregnancy was statistically different (P < 0.001) from excretion during the nonpregnancy period. Baseline progestogen concentrations were different from pregnancy (P < 0.001) and postovulatory (P < 0.01) concentrations, and progestogen concentrations during pregnancy period were different (P < 0.001) from postovulatory concentrations. In the nontreated cheetah (ZGT-3301), basal and increased progestogen concentrations were statistically different (P < 0.01). On the basis of 17beta-estradiol excretory patterns, duration of the estrous cycle (x +/- SEM) was 13.2 +/- 2.2 days. These results suggest that the enzyme-linked immunosorbent methods reported in this study were capable of quantifying reproductive hormones in fecal extracts of cheetahs and could be a practical alternative to other enzyme-linked immunosorbent assays which require more complex procedures.     

A sandwich enzyme immunoassay for bovine interferon-γ and its use for the detection of tuberculosis in cattle

An in vitro cellular assay for bovine tuberculosis has recently been developed. This assay detects gamma-interferon released in response to specific antigen in a whole blood culture system. The bio-assay previously described for the detection of bovine gamma-interferon (IFN-gamma) has now been replaced with a sandwich enzyme immunoassay (EIA) which utilises two monoclonal antibodies to bovine IFN-gamma. The EIA detects less than 25pg/ml of recombinant bovine IFN-gamma and is specific for biologically active bovine IFN-gamma; and does not detect bovine alpha or beta interferon. IFN-gamma from sheep, goat and buffalo, but not from pig, deer or man, are also recognised by the EIA. The bovine IFN-gamma EIA when used in conjunction with the whole blood culture system has resulted in a simple, rapid and sensitive in vitro assay for specific cell mediated immune responsiveness to M. bovis infection in cattle.     

Effects of Lepidium meyenii Walp and Jatropha macrantha on blood levels of estradiol-17 beta, progesterone, testosterone and the rate of embryo implantation in mice

The effects of two Peruvian folk medicines, Lepidium meyenii Walp and Jatropha macrantha, on mouse sex steroid hormones and embryo implantation were investigated. Progesterone levels increased significantly in mice that received L. meyenii Walp, while testosterone levels increased significantly in mice that received L. meyenii Walp as well as in those that received both L. meyenii Walp and J. macrantha. However, there were no marked changes in blood levels of estradiol-17beta or the rate of embryo implantation.     

The quantitative determination of fibrinogen in normal bovine plasma and in cows with inflammatory conditions

The quantitative determination of fibrinogen in normal plasma and in cows with inflammatory conditions.A rapid method for the quantitative determination of fibrinogen in bovine plasma is described.The method was employed in the determination of normal values in a material consisting of 100 cows and 50 calves and young animals of various ages. The mean value of the groups of cows was approximately 0.550 g/100 ml. For young animals it was somewhat lower and for cows in the last month of gestation moderately higher than in the other groups.The last part of the experiment involves the determination of the fibrinogen and γ-globulin levels in the plasma of 28 hospitalized cows with various inflammatory conditions. Group A in the material contained animals which were clinically cured and Group B animals that died or were killed.Both groups showed a considerable increase in the fibrinogen level. In Group A the mean value fell back to approximately the normal range while in Group B it remained constantly elevated.The sedimentation rate, SR, in human blood is primarily influenced by the fibrinogen content of the plasma. The SR in bovine blood is very low, and the test is therefore of little significance in diagnostic work. In conclusion, the possibility of using the fibrinogen determination in cattle for the same purpose as the SR in human blood is discussed.     

Development of a time-resolved fluorometry based immunoassay for the determination of canine haptoglobin in various body fluids

A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of haptoglobin (Hp) in canine serum. Haptoglobin was purified from canine acute phase serum by ammonium sulphate precipitation followed by gel filtration. This isolated dog Hp was used as the standard to calibrate the assay. Intra- and inter-assay coefficients of variation of the assay were, respectively, 5.7% and 16.6% at 0.51 mg/mL, 2.4% and 10.6% at 2.1 mg/mL and 10.5% and 11.9% at 32.5 mg/mL. The dilution of serum samples with high Hp concentrations resulted in linear regression equations with R2 of 0.99 and 0.97. A high correlation was found in serum Hp measurements by TR-IFMA and a commercial assay based on peroxidase activity of haemoglobin bound to haptoglobin (R2 = 0.96). The limit of detection for the TR-IFMA method was 0.002 microg/mL. The addition of fresh haemolysate to serum samples did not affect the haptoglobin concentration (P = 0.694). Statistical differences (P < 0.003) were found between healthy dogs and dogs with different pathological processes. In whole blood, Hp concentrations were much lower than in serum but closely related (R2 = 0.84) whereas saliva Hp concentrations were poorly related with serum concentrations (R2 = 0.53). However, the concentration of Hp in saliva was significantly (P < 0.039) higher in dogs with pathological processes compared to healthy dogs. The assay sensitivity was adequate to also be applied to whole blood and saliva specimens.     

The effect of beta carotene supplementation on the beta carotene and vitamin A levels of blood plasma and some fertility indices of dairy cows

Seasonal variations in beta carotene intake and low levels of beta carotene and vitamin A in blood plasma were found in cows fed ration based mainly on maize silage. The supplementation of daily winter rations with 300 mg Rovimix-beta carotene per cow, beginning 14 days before parturition and 60 days after calving increased slighty the beta carotene and vitamin A concentrations of blood plasma and improved some fertility indices: the number of inseminations per cow was reduced and the percentage of conception rate was significantly higher. It may be assumed that beta carotene content in feeds and it's utilization rate generally reflected in beta carotene blood plasma level and in the improvement of fertility.