Serum progesterone determination as an aid for pregnancy diagnosis in goats bred out of season

Nubian does (n = 12) were bred by artificial insemination after induction of estrus with medroxyprogesterone acetate impregnated vaginal sponges and pregnant mare serum gonadotrophin injections during the anestrous season. Pregnancy status was predicted from serum samples collected 21 days following the last breeding and analyzed using 1) a commercial bovine milk progesterone enzyme immunoassay test (EIA), and 2) a radioimmunoassay progesterone (RIA) test. Both tests detected nonpregnancy (EIA 100%, RIA 80%) more accurately than pregnancy (EIA 66%, RIA 75%). Commercial bovine progesterone EIA kits have potential as rapid, inexpensive screening tests for nonpregnant does bred out of season.     

Progesteronbestimmung in Kuhmilch: Vergleich der Ergebnisse des RIA-Milchfett- und EIA-Direkttests sowie eine Qualitätskontrolle des EIA-Direkttests

Inhalt Durch zwei Versuche sollte geklärt werden, wie gut die Übereinstimmung bei der Milchprogesteronbestimmung mit den Meβverfahren RIA-Milchfettextraktion und EIA-Direkttest ist und welche Reproduzierbarkeiten beim EIA zu erreichen sind. Beim Vergleich der Oeiden Bestimmungsverfahren—die Proben wurden unabhängig voneinander in zwei getrennten Labors analysiert—ergab sich eine Korrelation von r = 0.86 (P < 0.01). Der Qualitätstest ergab für Proben mit sehr niedrigen Progesterongehalten Variationskoeffizieriten von intra-assay 27% und inter-assay 53%. Bei mittleren Hormonkonzentrationen betrugen die entsprechenden. Werte 10% und 19% und bei hohen Werten 10% und 14%. Die ermittelten Ergebnisse stellen die Eignung des enzymimmunologischen Dirckttests für die Progesteronbestimmung in der Milch zum Zwecke der routinemäβigen Fruclitbarkeitsüberwachung unter Beweis. Contents: Determination of progesterone in cow's milk; comparison of the RIA performed on fat extract with the EIA performed on unextracted milk and assessment of the reliability of the EIA Two experiments were conducted to assess whether an enzyme immunoassay (EIA) is suitable to measure the concentration of progesterone in cow's milk. In trial 1 progesterone was determined in whole milk with the EIA and in the fat extract of the same samples with a radioimmunoassay (RIA). Agreement of the two assays was very good; the correlation coefficient was r = 0.86 (P < 0.01). In trial 2 the repeatability of the milk progesterone determination with the EIA was assessed. Milk samples from five cows each at three levels of milk progesterone (low, medium, high) were used. Each sample was determined five times in duplicate on each of five successive days. The mean intra- and inter-assay coefficients of variation at the low, medium and high progesterone Ievels were 27% and 53%, 10% and 19%, 10% and 14%, respectively. In conclusion the enzyme immunoassay is suitable to monitor ovarian function in dairy cows by way of milk progesterone determination.     

Evaluation of an automated,homogeneous enzyme immunoassay for serum thyroxine measurement in dog and cat serum

A homogenous enzyme immunoassay (EIA) for measurement of serum thyroxine (T4) concentration was evaluated for use with canine and feline serum. The EIA method was linear from 0 to 150 nmol T4/L for human serum, 0 to 94 nmol T4/L for feline serum and 10 to 60 nmol T4/L for canine serum. Intra- and interassay precision studies yielded coefficients of variation 相似文献   

Measurement of progesterone in sheep using a commercial ELISA kit for human plasma

Determination of serum or plasma progesterone (P4) concentrations is important to recognize pregnant and non-pregnant ewes, and also to predict the number of carried lambs. The 2 most common methodologies for the detection of plasma P4 are radioimmunoassay (RIA) and enzyme immunoassay (EIA). RIA is very expensive, and not all laboratories are equipped to perform this test; EIA is commercially available for human use, but only a few companies produce species-specific kits, which are expensive. We verified for ovine plasma a less expensive and easily available ELISA kit (DiaMetra) designed to quantify P4 in humans. Pools of ovine and human plasma were used to compare repeatability, accuracy, sensitivity, and stability of P4 measured by the DiaMetra kit. Repeatability data were within 15%, and accuracy values were ~90% for both plasma matrices. Stability data showed a loss of <20% for freeze–thaw and <30% for 30-d storage. All parameters were acceptable under international guidelines for method validation. The human ELISA kit was used successfully to quantify plasma P4 in 26 ewes during pregnancy until delivery. P4 concentrations were also correlated with the number of carried lambs.     

Pregnancy status determination in mares using a rapid lateral flow test for measuring serum oestrone sulphate

AIMS: To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point. METHODS: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visible detection reagent. Concentrations of the coating antigen and OS monoclonal antibody were optimised so that the working range would allow pregnancy status to be determined from a visual end-point. The test was run by adding 0.1 ml serum to the sample well of a plastic cassette encasing the test membrane. As the serum migrated along the membrane, a test dot and control line were generated on it within 5-10 min. The intensity of the test dot was inversely proportional to the concentration of OS in the serum sample being tested. Results were compared with those from a validated OS enzyme immunoassay (EIA) and subsequent foaling or return to oestrus of the mares. RESULTS: Serum samples with OS concentrations <10 ng/ml, indicative of non-pregnancy in mares >100 days post-mating, generated a test end-point consisting of a highly visible test dot and control line, whereas serum OS concentrations >50 ng/ml, indicative of pregnancy, generated a control line only. The test correctly identified 384/389 (98.7%) non-pregnant mares tested, and 303/312 (97.1%) pregnant mares tested that were >100 days post-mating. The lateral flow test devices were stable for at least 12 months when stored at 4 degrees C, sealed in aluminium pouches with desiccant. CONCLUSION: This novel, rapid, easy-to-use, lateral flow immunoassay offers a practical alternative to traditional laboratory- based immunoassays for measuring serum OS concentrations in mares for determining their pregnancy status.     

Long-term cannulation of the ovarian vein in mares.

A cannulation technique was developed to collect blood samples from the ovarian vein of mares over an extended period. Ovarian venous cannulae placed in 4 mares remained patent for a mean (+/- SEM) duration of 36.8 (+/- 6.2) days. During mid-diestrus, concentrations of progesterone in the ovarian vein ipsilateral to the corpus luteum (1,663.8 +/- 238.8 ng/ml) were significantly (P less than 0.001) higher than concentrations measured in paired samples from the jugular vein (6.1 +/- 0.3 ng/ml). Concentration of estradiol in the ovarian vein ranged from a mean of 1,053.2 +/- 303.1 pg/ml during diestrus to 3,353.8 +/- 1,052.8 pg/ml during estrus, whereas values for 74% of samples collected concurrently from the jugular vein were near or below the sensitivity of the assay (10 pg/ml). Results of the study indicate that patent long-term ovarian vein cannulation can be achieved in mares. The cannulation technique provides access to important fundamental information on equine reproductive endocrinology, which to our knowledge, has not been available.     

Development and evaluation of a rapid enzyme-immunoassay system for measurement of the urinary concentration of estrone-3-glucuronide in a female giant panda (Ailuropoda melanoleuca)

To detect estrus for reproductive management, and to determine the relationship between urinary estrogen and estrous behavior, in a female giant panda, we developed and evaluated a rapid enzyme immunoassay (EIA) system for urinary Estrone-3-glucuronide (E1G) using commercial reagents. The developed EIA system took only around 3 hours, including all procedures to obtain a result. It indicated good reproducibility (intra-assay CV of 5.16%, interassay CV of 15.4%) and sensitivity (lowest standard concentration was 0.0156 ng/ml) for measurement of the urinary concentrations of E1G in the giant panda. There was a positive correlation (r=0.934) with the data for estrone (E1) in the same samples, as measured by radioimmunoassay (RIA) performed in a commercial laboratory. The changes in the E1G concentrations were almost synchronous with the changes in E1 assayed by RIA in urine collected during 4 consecutive estrous seasons. The dynamics of urinary E1G measured by this system highly correlated with the occurrence of the presenting estrous behavior in the giant panda. The above results indicate that this assay system may be normally, rapidly and practically used for measurement of the urinary concentration of E1G in the giant panda.     

Effect of repeated administration of oxytocin during diestrus on duration of function of corpora lutea in mares

OBJECTIVE: To determine whether IM administration of exogenous oxytocin twice daily on days 7 to 14 after ovulation blocks luteolysis and causes prolonged function of corpora lutea (CL) in mares. DESIGN: Prospective study. ANIMALS: 12 mares. PROCEDURES: Beginning on the day of ovulation (day 0), jugular blood samples were collected every other day until day 40 for determination of progesterone concentration. On day 7, mares (n = 6/group) were treated with saline (0.9% NaCl) solution (control group) or oxytocin. Beginning on day 7, control mares received 3 mL of sterile saline solution every 12 hours, IM, and oxytocin-treated mares received 60 units of oxytocin every 12 hours, IM, through day 14. Mares were considered to have prolonged CL function if progesterone concentration remained > 1.0 ng/mL continuously through day 30. RESULTS: The proportion of mares with prolonged CL function was significantly higher in the oxytocin-treated group (6/6), compared with the control group (0/6). All control mares underwent luteolysis by day 16, at which time their progesterone concentrations were < 1.0 ng/mL. In contrast, all 6 oxytocin-treated mares maintained progesterone concentrations > 1.0 ng/mL continuously through day 30. CONCLUSIONS AND CLINICAL RELEVANCE: IM administration of 60 units of oxytocin twice daily on days 7 to 14 after ovulation was an efficacious method of inhibiting luteolysis and extending CL function in mares. Disrupting luteolysis by administering exogenous oxytocin during diestrus appears to be a plausible and practical method of long-term suppression of estrus in mares.     

Follicle stimulating hormone, luteinising hormone and progesterone concentrations in the blood of thoroughbred mares exhibiting single and twin ovulations

The concentrations of follicle stimulating hormone, luteinising hormone and progesterone were measured in serial blood samples taken throughout one or more oestrous cycles from 12 Thoroughbred mares, some of which exhibited single and others twin ovulations. The resulting profiles clearly demonstrated that no simple relationship exists between circulating gonadotrophin levels and subsequent ovulation rate in the mare. However, plasma progesterone concentrations during dioestrus are, as expected, higher following twin than single ovulations. The findings suggest that the underlying cause of twin ovulation in some mares may exist at the ovarian end of the pituitary-ovarian axis which controls follicular development and ovulation.     

Pregnancy status determination in mares using a rapid lateral flow test for measuring serum oestrone sulphate

AIMS: To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point.

METHODS: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visible detection reagent. Concentrations of the coating antigen and OS monoclonal antibody were optimised so that the working range would allow pregnancy status to be determined from a visual end-point. The test was run by adding 0.1 ml serum to the sample well of a plastic cassette encasing the test membrane. As the serum migrated along the membrane, a test dot and control line were generated on it within 5–10 min. The intensity of the test dot was inversely proportional to the concentration of OS in the serum sample being tested. Results were compared with those from a validated OS enzyme immunoassay (EIA) and subsequent foaling or return to oestrus of the mares.

RESULTS: Serum samples with OS concentrations <10 ng/ml, indicative of non-pregnancy in mares >100 days post-mating, generated a test end-point consisting of a highly visible test dot and control line, whereas serum OS concentrations >50 ng/ml, indicative of pregnancy, generated a control line only. The test correctly identified 384/389 (98.7%) non-pregnant mares tested, and 303/312 (97.1%) pregnant mares tested that were >100 days post-mating. The lateral flow test devices were stable for at least 12 months when stored at 4°C, sealed in aluminium pouches with desiccant.

CONCLUSION: This novel, rapid, easy-to-use, lateral flow immunoassay offers a practical alternative to traditional laboratory-based immunoassays for measuring serum OS concentra- tions in mares for determining their pregnancy status.     

Comparison of glucose concentrations in canine whole blood,plasma, and serum measured with a veterinary point-of-care glucometer

Previous studies have determined that, compared to whole blood, serum or plasma used in a portable blood glucometer (PBG) may provide more accurate results. We investigated the accuracy of a veterinary PBG (AlphaTRAK 2; Zoetis) for the measurement of glucose concentrations in serum, plasma, and whole blood compared to plasma glucose concentration measured by a biochemical analyzer. Blood samples from 53 client-owned dogs were collected. Lin concordance correlation coefficient (ρ_c) and Bland–Altman plots were used to determine correlation and agreement between the results obtained for the different sample types. Glucose concentration in whole blood measured by the veterinary PBG was more strongly correlated with the glucose concentration measured by the biochemical analyzer (ρ_c = 0.92) compared to plasma and serum glucose concentrations (ρ_c = 0.59 and 0.57, respectively). The mean differences between the glucose concentrations in whole blood, plasma, and serum measured by the veterinary PBG and the glucose concentration determined by the biochemical analyzer were 1.0, 6.3, and 6.7 mmol/L (18, 113, and 121 mg/dL), respectively. Our findings suggest that, when using this veterinary PBG, the accuracy of a glucose measurement obtained is higher when using whole blood compared to plasma or serum. Use of whole blood allows for more correct assessment and diagnosis, which are necessary for appropriate therapeutic intervention.     

A Retrospective Analysis of 2,253 Cases Submitted for Endocrine Diagnosis of Possible Granulosa Cell Tumors in Mares

Endocrine diagnoses of granulosa cell tumors (GCTs) in the mare are frequently based upon determination of serum concentrations of inhibin, testosterone, and progesterone (GCT panel). In the present study, we examined results from 2,253 samples submitted for determination of GCT panel. In an additional subset of samples (n = 75), diagnosis of GCT was confirmed based upon surgery or necropsy. The objective of the current study was to examine the agreement between serum inhibin and testosterone concentrations in mares with serum progesterone concentrations less than or ≥1 ng/mL. Across all samples, elevations in serum inhibin or testosterone were noted in 15.2% or 17.3% of samples, respectively. For samples with progesterone <1 ng/mL, more (P = .001) samples had elevated inhibin than elevated testosterone, whereas for samples with progesterone ≥1 ng/mL, more (P < .0001) samples had elevated testosterone than elevated inhibin. For samples with progesterone <1 ng/mL, the agreement between elevated inhibin and testosterone was greater than that for samples with progesterone ≥1 ng/mL. In a subset of 53 samples with progesterone ≥1 ng/mL, 28 samples had endocrine evidence of a granulosa cell tumors based upon elevations of serum anti-Müllerian hormone, and 4 of these samples had endocrine evidence of pregnancy based upon estrone sulfate concentrations. For samples from mares with confirmed GCTs, 85% and 55% had elevated inhibin or testosterone, respectively. Based upon endocrine diagnosis, GCTs occur in mares with elevated progesterone and/or estrone sulfate, albeit at a low rate.     

Comparison of 2 enzyme immunoassays and a radioimmunoassay for measurement of progesterone concentrations in bovine plasma, skim milk, and whole milk

The objective of this study was to compare 2 enzyme immunoassays (EIAs) with a radioimmunoassay (RIA) as to sensitivity and accuracy in the measurement of the progesterone (P4) concentration in bovine plasma, skim milk, and whole milk. The 72 samples from 24 lactating dairy cows expected to have either a high P4 concentration (cows in diestrus or pregnant) or a low P4 concentration (cows in estrus or anestrus) were analyzed by RIA, solid-phase EIA (SPEIA), which included a solvent extraction step, or direct EIA (DEIA) without solvent extraction. The overall mean concentrations of P4 did not differ (P < 0.4) among the assays. However, for the cows that were in diestrus or pregnant, the mean P4 concentrations (and standard error) were higher (P < 0.03), regardless of sample type, with RIA than with SPEIA, at 7.3 (0.7) and 6.1 (0.6) ng/mL, respectively. When only the high-P4 samples analyzed by RIA were compared, the mean P4 concentration was higher (P < 0.001) in whole milk than in skim milk, at 9.8 (1.0) and 4.1 (0.7) ng/mL, respectively. Although the mean P4 concentrations in the low-P4 samples did not differ (P < 0.80) among assays, the proportions of cows with a P4 concentration > or = 1 ng/mL were 3%, 14%, and 44% for RIA, SPEIA, and DEIA, respectively (P < 0.01; DEIA > SPEIA > RIA).     

Prolactin concentrations are not suppressed in mares administered constant exogenous melatonin

On the summer solstice (June 21, 1996), six of 12 intact light horse mares randomly chosen from a larger herd were subcutaneously implanted with ALZET^®a osmotic minipumps containing a melatonin solution (16 mg/ml) designed to release approximately 960 μg of melatonin/day. An additional two mares received implants containing only the saline-DMSO vehicle and four remained untreated. Blood samples were collected on days 5, 26, and 59 of treatment to monitor melatonin concentrations and to verify pump function. Prolactin concentrations were determined from blood samples collected via jugular cannulae every 12 min for 8 hours on days 25,46, and 89 after initial implantation. On day 89, samples were collected hourly for 16 hours following the initial 8-hour sampling period. Melatonin and prolactin concentrations were determined in the blood samples by radioimmunoassay. Mean circulating concentrations of melatonin in treated mares (n=6) were found to be significantly elevated when compared to controls (n=6); however, there was no significant difference in prolactin concentrations between the groups. These studies demonstrate that longterm treatment with melatonin is unaccompanied by a change in prolactin secretion.     

Endocrine Profile Comparisons of Fat Versus Moderately Conditioned Mares Following Parturition

The influence of progesterone, luteinizing hormone (LH), and estrogen on the mare's estrous cycle has been well researched and documented, but other endocrine profiles have not received as much attention. To evaluate endocrine concentrations in fat-conditioned (body condition score [BCS] of 7–8) versus moderately conditioned mares (BCS of 5–6), 24 mares were allotted to and maintained in respective groups from late gestation until pregnancy was confirmed after breeding on the second postpartum estrus. Serum concentrations of thyroxine (T₄), insulin-like growth factor-1 (IGF-1), and leptin were assayed to characterize circulating blood concentrations. Additionally, LH and progesterone serum concentrations were assayed to evaluate the estrous cycle status of the mares. Leptin and progesterone concentrations were not different (P > .05) between the groups. Nevertheless, serum concentrations of T₄ were higher (P < .01) and IGF-1 concentrations lower (P < .01) in moderately conditioned as compared with fat-conditioned mares during times of ovulation and the interovulatory period. Furthermore, serum concentrations of LH were found to be different between the groups only when the estrous cycle approached the second ovulation (P < .0001). Results of this study suggest that mares maintained in a BCS of 5 or greater are similar in terms of reproductive efficiency. Although the circulating serum concentrations of T₄ and IGF-1 are different after parturition, their influence does not affect reproductive capabilities of mares with a BCS of 5 or greater.     

The effect of pulsatile gonadotropin-releasing hormone and estradiol administration on luteinizing hormone and follicle-stimulating hormone concentrations in pituitary stalk-sectioned ovariectomized pony mares

Hourly pulses of gonadotropin-releasing hormone (GnRH) or bi-daily injections of estradiol (E2) can increase luteinizing hormone (LH) secretion in ovariectomized, anestrous pony mares. However, the site (pituitary versus hypothalamus) of positive feedback of estradiol on gonadotropin secretion has not been described in mares. Thus, one of our objectives involved investigating the feedback of estradiol on the pituitary. The second objective consisted of determining if hourly pulses of GnRH could re-establish physiological LH and FSH concentrations after pituitary stalk-section (PSS), and the third objective was to describe the declining time trends of LH and FSH secretion after PSS. During summer months, ovariectomized pony mares were divided into three groups: Group 1 (control, n = 2), Group 2 (pulsatile GnRH (25 μg/hr), n = 3), and Group 3 (estradiol (5 mg/12 hr), n = 3). All mares were stalk-sectioned and treatment begun immediately after stalk-section. Blood samples were collected every 30 min for 8 h on the day before surgery (DO) and 5 d post surgery (D5) to facilitate the comparison of gonadotropin levels before and after pituitary stalk-section. Additionally, jugular blood samples were collected every 12 hr beginning the evening of surgery, allowing for evaluation of the gonadotropin secretory time trends over the 10 d of treatment. On Day 10, animals were euthanized to confirm pituitary stalk-section and to submit tissue for messenger RNA analysis (parallel study). Plasma samples were assayed for LH and FSH by RIA. Mean LH secretion decreased from Day 0 to Day 5 in Groups 1 and 3, whereas LH secretion tended (P < 0.08) to decrease in Group 2 mares. On Day 5, LH was higher (P < 0.01) in Group 2 (17.26 ± 3.68 ng/ml; LSMEANS ± SEM), than either Group 1 (2.65 ± 4.64 ng/ml) or group 3 (4.28 ± 3.68 ng/ml). Group 1 did not differ from Group 3 on Day 5 (P < 0.40). Similarly, mean FSH levels decreased in all groups after surgery, yet Group 2 mares had significantly (P < 0.001) higher FSH concentrations (17.66 ± 1.53 ng/ml) than Group 1 or Group 3 (8.34 ± 1.84 and 7.69 ± 1. 63 ng/ml, respectively). Regression analysis of bi-daily LH and FSH levels indicated that the time trends were not parallel. These findings indicate: 1) Pituitary stalk-section lowered LH and FSH to undetectable levels within 5 d after surgery, 2) pulsatile administration of GnRH (25 μg/hr) maintained LH and FSH secretion, although concentrations tended to be lower than on Day 0, and 3) E2 did not stimulate LH or FSH secretion.     

Reversibility of the effects of GnRH‐vaccination used to suppress reproductive function in mares

Reasons for performing study: Active immunisation against gonadotrophin‐releasing hormone (GnRH) provides a reversible method for control of oestrous behaviour and fertility in mares. Previous reports failed to demonstrate the interval to resumption of cyclic ovarian activity after GnRH‐vaccination. Hypothesis: Administration of the GnRH‐vaccine Improvac in a large group of mares of various ages will result in effective, reliably reversible suppression of ovarian activity within a 2 year period. Methods: The mares, subdivided into 3 age categories, were vaccinated twice (with a 35 day interval) using 400 µg Improvac and monitored via blood samples until Day 720 after initial vaccination for serum progesterone concentration determination by radioimmune assay and anti‐GnRH antibody titre by enzyme immunoassay. Samples were collected until individuals resumed cyclic ovarian activity. Results: All mares showed suppression of cyclic ovarian activity by clinical examination and serum progesterone concentration (SPC) ≤1 nmol/l by Day 70 and 92.2% resumed cyclic activity by SPC at Day 720 with a mean interval = 417.8 days (s.d. ± 23.9; range 232–488 days, median 344 days). A significant age effect (P = 0.028) on the interval, but not on GnRH‐antibody titre response, was observed between the youngest (≤4 years) and oldest (≥11 years) categories. Conclusions: Immunising adult mares of all ages with Improvac resulted in a reversible suppression of cyclic ovarian activity in most mares. An age effect, with the youngest mares showing a longer interval to reversibility, was observed.     

Endocrine profiles of periparturient mares and their foals

The aim of this study was to characterize concentrations of leptin, IGF-I, and thyroid stimulating hormone (TSH) in the blood serum of mares pre-and postpartum, in the milk serum of mares postpartum, and in the blood serum of their foals. Nine pregnant Quarter Horse mares and their offspring were used in this study. Once weekly between 1000 and 1200 h for 2 wk before their predicted parturition date, mares were weighed, assigned a BCS, and blood was sampled via jugular venipuncture. Within 2 h of parturition and before the foals nursed (d 0), blood samples were obtained from the mares and foals, and a milk sample was collected from the mares. Blood from the foals and blood and milk from the mares were collected again at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 12, 19, 26, 33, and 61 d postpartum. Mares and foals also were weighed and assigned a BCS on d 0, 5, 12, 19, 26, 33, and 61. Additionally, on d 5, 33, and 61, ultrasound images of fat depth and area of the LM immediately cranial to and parallel with the last rib on the left side of the foals were measured to characterize changes in fat depth and LM area over time. There were no changes in mare blood concentrations TSH (P = 0.15), nor were there any changes in foal blood concentrations of leptin (P = 0.54) or TSH (P = 0.10) during the trial period. Mare blood concentrations of IGF-I tended to change over time (P = 0.07), whereas leptin changed over time (P < 0.001), initially decreasing and then remaining relatively stable after d 5. Foal blood concentrations of IGF-I increased initially, peaked at d 19, and stabilized thereafter (P < 0.001). Milk concentrations of leptin and TSH were greatest on d 0 and decreased over time (P < 0.007), reaching nadir concentrations at d 61. Milk concentrations of IGF-I also changed over time (P = 0.02), being greatest on d 0 and undetectable by d 12. There was no difference in BCS (P = 0.94) in mares over time, but there was a difference between pre- and postpartum BW (P < 0.001) due to foaling. However, no differences were detected in pre- (P = 0.70) or postpartum BW (P = 0.76) of mares over time. Mean ultrasonic fat depth and LM area increased (P < 0.04) as the foals aged, as did BCS and BW (P < 0.001). Recognizing changes in metabolic hormones surrounding the time of parturition in the mare and foal provides a basis for further determination of the role, if any, these hormones play in the milk, as well as in the neonate.     

Testosterone administration to mares during estrus: duration of estrus and diestrus and concentrations of LH and FSH in plasma

To study the possible role of ovarian androgens in regulation of follicle stimulating hormone (FSH) secretion in the cycling mare, five mature, intact mares were treated with testosterone (20 micrograms/kg of body weight) daily during estrus; five control mares received safflower oil on the same schedule. Mares were teased for estrus and samples of jugular blood were drawn daily through one full estrous cycle. Concentrations of FSH in plasma were measured by a newly developed radioimmunoassay based on anti-ovine FSH serum and radioiodinated equine FSH. Testosterone treatment during estrus had no effect on duration of estrus, diestrus or the total cycle. Concentrations of FSH in plasma during estrus were unaffected by testosterone treatment. However, FSH concentrations in testosterone-treated mares were elevated (P less than .05) compared with controls during mid-diestrus (d 6 through 11). The magnitude and timing of the LH peaks were unaffected by treatment, as was the day on which the first elevated progesterone concentration occurred. These data are consistent with a model of FSH secretion in which ovarian androgens cause an accumulation of FSH in the pituitary during estrus in preparation for the surges that occur in FSH secretion during diestrus.     

In vitro metabolism of ovarian steroids by bovine whole blood and plasma

Experiments were conducted to evaluate the effects of time and temperature on the potential of bovine whole blood (WB) or plasma (PL) to metabolize the ovarian steroids progesterone, estradiol-17β and testosterone. During a radioimmunoassay study (Experiment 1), we observed a temperature and time-dependent reduction (P<0.001) of plasma progesterone concentrations in samples incubated as WB at 5, 15, 25, or 35C for up to 48 hr. Most notable was the observation that 27% of progesterone present in controls was lost when WB was incubated at 5C for 48 hr and a 17% reduction was observed when PL samples were incubated at 35C for 48 hr. Immunoreactive estradiol-17β concentrations (Experiment 2) in PL and WB incubates were not affected by time or temperature. However, immunoreactive testosterone concentrations increased more than 3-fold by 48 hr in WB incubates held at 35C. To examine the latter observation further, ³H-progesteone was incubated with WB at 35C, followed by extraction and thin-layer chromatography (Experiment 3). Results generally supported RIA findings and revealed the presence of significant 17α-hydroxylase, 17–20 lyase and aromatase activity. Heretofore this has not been considered to occur outside major steroid metabolizing organs.