Effects of induced endometritis on uterine blood flow in cows as evaluated by transrectal Doppler sonography

This study was conducted to evaluate the effects of induced endometritis on uterine blood flow in cows. Transrectal Doppler sonography was performed on uterine arteries of six cyclic cows before and for 4 days after inducing acute endometritis by intrauterine infusion of 720 mg of policresulen, and for 4 days of the following estrous cycle. Time-averaged maximum velocity (TAMV) increased (p < 0.001) and pulsatility index (PI) decreased (p < 0.0001) within 1 h of policresulen administration, and did not change (p > 0.05) in the next 4 days of the same cycle. TAMV and PI values in the subsequent cycle did not differ (p > 0.05) from the values measured before infusion and showed no changes (p > 0.05) within the cycle. Blood flow parameters were not related (p > 0.05) to plasma concentrations of progesterone and estrogen. All cows showed an acute endometritis determined by histopathological findings of biopsy samples taken 1 day after infusion and fibrotic endometrial alterations detected in the subsequent cycle. No relationships were observed between fibrotic changes of the endometrium and uterine blood flow during either cycle. In conclusion, acute inflammation is accompanied by a rise in uterine blood flow, but fibrotic alterations do not seem to be related to Doppler sonographic findings.     

Use of ultrasonic Doppler waveforms to estimate changes in uterine artery blood flow and vessel compliance

Previous data from our laboratory have demonstrated that uterine blood flow (UBF) and uterine arterial smooth muscle tone vary regularly during the estrous cycle of the cow. Uterine blood flow is highest and uterine arterial tone is lowest at estrus, whereas UBF is lowest and uterine arterial tone is highest during the luteal phase of the cycle. Blood flow through arteries is highly pulsatile; changes in arterial wall properties affect shape of the velocity waveform. This study was conducted to evaluate changes in uterine arterial velocity waveforms throughout the estrous cycle of the cow and to relate these changes to fluctuations in UBF and concentrations of estrogen and(or) progesterone in systemic blood. Pulsatile velocity waveforms were obtained daily from pulsed-wave ultrasonic probes placed surgically on the middle uterine artery of five beef cows exhibiting estrous cycles of normal duration (d 0 = day of estrus). Velocity waveforms varied regularly during the estrous cycle of each cow in association with changes in UBF and steroid concentrations. Further, two distinct velocity waveform shapes were observed during the estrous cycle. The first waveform shape, which was observed during periods of high UBF (d -4 to +4 of the estrous cycle), was characteristic of a highly compliant vessel and was associated with a high estrogen:progesterone ratio. The second waveform shape, which was observed from d 7 to 14 of the estrous cycle, was characteristic of a less compliant vessel and was associated with a depressed estrogen:progesterone ratio. These data suggest that compliance of the uterine artery changes during the estrous cycle in association with the changing estrogen:progesterone ratio in blood.     

Estrus, ovulation, and serum progesterone, estradiol, and LH concentrations in mares after an increased photoperiod during winter

On December 11, 1974, 15 seasonally anestrous mares were assigned at random to 1 of 3 experimental groups: outdoor-control, indoor-control, or indoor light-treated (a 16-hour photo-period). This experiment was terminated on April 21, 1975. The five mares in the indoor light-treated group ovulated 59.0+/-6.9 days later, which was 74 days earlier (P less than 0.01) than 2 of the 5 outdoor-controls (the other 3 ovulated after April 21 during a subsequent experiment) and 50 days earlier (P less than 0.05) than the indoor-controls. Durations of the 1st estrus for the 3 groups of mares were 13.3+/-3.6, 8.4+/-2.0, and 6.0+/-1.0 days for the indoor light-treated, indoor-control, and outdoor-control groups, respectively. The indoor light-treated mares averaged 4.2 estrous cycles before April 21, the indoor-control mares averaged 1.4 estrous cycles, and 2 of 5 outdoor-control mares ovulated 1 time during the experiment. The peripheral blood luteinizing hormone (LH), estradiol, and progesterone concentrations were minimal during winter anestrous. The hormone changes normally associated with estrous cycle activity in mares--maximal estradiol and luteinizing hormone concentrations near ovulation and maximal progesterone concentration during diestrus--were observed in all mares beginning at the 1st estrus. Hair loss was observed earlier in the light-treated mares, than in either of the other groups. In conclusion, a 16-hour photo-period initiated in early December for anestrous brood mares caused endocrinologically normal estrous cycles to begin within 2 months. This may allow breeding and foaling considerably earlier than normally expected.     

Uterine clearance mechanisms during the early postovulatory period in mares

Uterine response to inoculation with Streptococcus zooepidemicus organisms, 51Cr-labeled 15-microns microspheres, and charcoal was evaluated in 9 mares (4 resistant and 5 susceptible to endometritis) to determine mechanical and cellular clearance rates during the early postovulatory period. Mares were inoculated at estrus prior to ovulation during estrous cycles 1, 3, and 5. Uterine swab specimens for aerobic and anaerobic bacteriologic culture and serum for progesterone determination were obtained on postovulation day 3 during estrous cycle 1, on the day of ovulation during estrous cycle 3, and on postovulation day 5 during estrous cycle 5. Immediately thereafter, the uterus was irrigated with 50 ml of sterile physiologic saline solution containing tracer amounts of 125I-labeled human serum albumin. Streptococcus zooepidemicus was isolated from 10 of 15 (67%) uterine specimens collected from susceptible mares and incubated aerobically. Escherichia coli also was isolated from 2 of the 10 specimens incubated aerobically. Anaerobic bacteriologic culture of specimens from all mares yielded no growth. Chromium-labeled microspheres were recovered twice from 2 susceptible mares, on day 0 and day 5. Charcoal was retained in 5 specimens collected from 3 susceptible mares. Bacteriologic culture of specimens from resistant mares did not yield growth. On day 0, chromium-labeled microspheres and charcoal were recovered once from 1 resistant mare. Mares susceptible to endometritis accumulated more fluid within the uterine lumen after ovulation than did resistant mares (mean +/- SEM, 52.73 +/- 15.22 ml and 7.41 +/- 1.96 ml, respectively; P less than 0.01). From this study, it appeared that uterine cellular and bactericidal mechanisms are dysfunctional during the early postovulatory period. However, there appeared to be no disruption of the mechanisms responsible for mechanical clearance of materials inoculated in the uterus.     

Microvascular development and growth of uterine tissue during the estrous cycle in mares

OBJECTIVE: To document uterine growth and microvascular development in the endometrium of uteri with differing degrees of fibrosis as well as uterine growth throughout the estrous cycle of mares. ANIMALS: 30 mares. PROCEDURE: Uterine tissue was obtained during the breeding season from a slaughter facility. Stage of estrous cycle of the mares was assessed on the basis of ovarian structures and plasma progesterone concentrations. Endometrium was characterized by use of light microscopy, and blood vessel walls were marked by histochemical techniques. Microvascular development was evaluated by a computerized image analysis system. Growth of uterine tissue was based on cellular content of DNA and RNA, RNA:DNA, and protein:DNA. RESULTS: Significant differences in vascular density were not observed in the endometrium of uteri obtained from mares euthanatized during the follicular or luteal phase of the estrous cycle, regardless of whether endometrial classification of degree of fibrosis was considered. There was a 3-fold increase in amount of DNA and RNA of endometrial cells in the follicular phase when compared to myometrium. Hypertrophy of endometrial tissue during the luteal phase was reflected by a significant increase in cell protein content and protein:DNA. CONCLUSIONS AND CLINICAL RELEVANCE: Endometrial growth of vascular tissues during the estrous cycle may be coordinated with development of nonvascular tissue. Estrogen and progesterone may play a role in regulation of uterine growth and angiogenesis.     

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.     

Effect of transportation on the estrous cycle and concentrations of hormones in mares

Effect of transportation on estrous behavior, duration of the estrous cycle, ovulation, pregnancy rates and concentrations of serum cortisol, plasma ascorbic acid (AA), LH, estradiol and progesterone in mares was investigated. Fifteen mares were transported for 792 km (12 h) during the preovulatory stage of estrus. Transported mares were bled immediately before transport (baseline), at midtrip and 0, 12, 24, 48 and 72 h post-transport and twice daily from d 1 before transport to d 1 (estrogen) or 3 (LH) post-ovulation. Blood samples also were taken for progesterone on d 0, 2, 6, 10, 15, 16, 17, 18, 19 and 20 post-ovulation. Nontransported control mares (n = 15) were bled on the same schedule as transported mares. There was no difference (P greater than .05) in number of mares ovulating, estrous behavior, duration of the estrous cycle or pregnancy rate between groups. Cortisol in transported mares increased to concentrations greater (P less than .05) than those in control mares at midtrip and 0 h post-transport. Concentrations of AA in transported mares also increased (P less than .05) at midtrip, then decreased (P less than .05) below baseline at 24 h post-transport. Concentrations of LH and estradiol increased (P less than .05) above baseline throughout the blood-sampling period. Increases apparently were due to preovulatory surges of these hormones. Increase in LH concentrations in transported mares, however, was greater (P less than .05) than that in control mares at 0 h post-transport.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression of progesterone receptor membrane component 1, serpine mRNA binding protein 1 and nuclear progesterone receptor isoforms A and B in the bovine myometrium during the estrous cycle and early pregnancy

The aim of this study was to investigate the (1) expression of progesterone membrane component 1 (PGRMC1), serpine mRNA binding protein 1 (SERBP1) and progesterone receptor (PR) mRNA and (2) protein expression levels of PGRMC1, SERBP1 and PR isoforms A and B in the bovine myometrium during the estrous cycle and early pregnancy. Uteri from cows on days 1-5, 6-10, 11-16 and 17-21 of the estrous cycle and weeks 3-5, 6-8 and 9-12 of pregnancy were used (n=5-6 per period). There were no changes (P>0.05) in PGRMC1 mRNA expression during the estrous cycle, while expression of SERBP1 and PR mRNA was the lowest (P<0.05) on days 11-16 relative to other days of the cycle. The highest mRNA expression of PGRMC1, SERBP1 and PR was found during pregnancy. There were no changes (P>0.05) in SERBP1 protein expression in cycling and pregnant cows, while the highest (P<0.05) PGRMC1 protein expression was found during weeks 3-5 of pregnancy. Similar protein expression profiles for PRA and PRB were found, and protein levels were highest on days 1-5 of the estrous cycle. From day 6 of the cycle, PRA and PRB protein expression decreased and were maintained at this lower level during pregnancy. In conclusion, our study assessed mRNA and protein expression levels of PGRMC1, SERBP1 and PR in the bovine myometrium during the estrous cycle and the first trimester of pregnancy. It is possible that progesterone (P4) affects myometrial function in a genomic and nongenomic manner.     

Progesterone receptor mRNA levels during pregnancy, labor, lactation and the estrous cycle in rat uterus

Progesterone plays important roles in the regulation of female reproduction. In this study, progesterone receptor (PR) mRNA levels in rat uterus during pregnancy, labor, lactation and the estrous cycle were examined by competitive RT-PCR. During pregnancy and lactation, PR mRNA levels had decreased on day 20 of pregnancy (P20) and P21 compared with P15 but increased during labor. After a decline on day 1 of lactation (L1), PR mRNA levels had increased again on L3 and L14 compared with P15, P18, P20, P21 and P21pm (at 2200-2300 h on P21). There was no significant change in the PR mRNA level during the estrous cycle. The PR mRNA level did not change during 1 week of progesterone treatment or afterwards. Injection of 17beta-estradiol did not affect PR mRNA levels in rats treated with progesterone or those without any injections. In rats on P18, 17beta-estradiol injection did not change PR mRNA levels after sham-operation but induced an increase in PR mRNA levels of rats ovariectomized 6 h before the treatment. These results suggest that uterine PR mRNA levels are differently regulated during late pregnancy, labor and lactation, and during labor estrogen is one of the essential factors for the increase in PR mRNA levels.     

Effect of deslorelin acetate on gonadotropin secretion and ovarian follicle development in cycling mares

OBJECTIVE: To evaluate gonadotropin secretion and ovarian function after administration of deslorelin acetate to induce ovulation in mares. DESIGN: Randomized controlled trial. ANIMALS: 16 healthy mares with normal estrous cycles. PROCEDURE: 8 control mares were allowed to ovulate spontaneously, whereas 8 study mares received deslorelin to induce ovulation when an ovarian follicle > 35 mm in diameter was detected. Follicle development and serum concentrations of gonadotropins were monitored daily during 1 estrous cycle. Pituitary responsiveness to administration of gonadotropin-releasing hormone (GnRH) was evaluated 10 days after initial ovulation. RESULTS: Interovulatory intervals of mares treated with deslorelin (mean +/- SD, 25.6 +/- 2.6 days) were longer than those of control mares (22.9 +/- 1.8 days). Diameter of the largest follicle was significantly smaller during 2 days of the diestrous period after ovulation in deslorelin-treated mares than in control mares. Concentrations of follicle-stimulating hormone (FSH) were lower in deslorelin-treated mares on days 5 through 14 than in control mares. Concentrations of luteinizing hormone were not different between groups during most of the cycle. Gonadotropin release in response to administration of GnRH was lower in mares treated with deslorelin acetate than in control mares. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of deslorelin was associated with reduction in circulating concentrations of FSH and gonadotropin response to administration of GnRH during the estrous cycle. Low concentration of FSH in treated mares may lead to delayed follicular development and an increased interovulatory interval.     

Ovarian, uterine and embryo dynamics in horses versus ponies

Characteristics of the internal genitalia were compared between horses and ponies contemporaneously in a single study by asingle operator. Diameter of the preovulatory follicle for 10 days before ovulation (Day 0) and cross-sectional area of the corpus luteum for 10 days after ovulation were measured in 12 horses and 12 ponies. Follicle growth rate and maximum diameter attained during growth and luteal tissue area were not different between mare types. However, ultrasonic echogenicity of luteal tissue was less on Day 0 (P<0.09) and Day 1 (P<0.01) in ponies than in horses, suggesting that under field conditions mare type should be considered when using the extent of luteal gland echogenicity for estimating the number of days postovulation and luteal function. Circulating concentrations of progesterone averaged over Days 3 to 10 were greater in ponies (10.3 ng/ml) than in horses (8.7 ng/ml) as suggested by a tendency (P<0.1) towards a type-by-day interaction.Luteal, uterine and embryonic characteristics during Days 0 to 40 of pregnancy were compared between 7 horses and 9 ponies. Luteal tissue area of the primary corpus luteum was not different between mare types during Days 0 to 30 and during luteal resurgence after Day 30. The interaction between mare type and day was significant for circulating progesterone concentrations; the interaction seemed to result primarily from higher progesterone levels in ponies during Days 6 to 12 and Days 34 to 40. Progesterone concentrations were about 25% higher in ponies on the approximate days that levels were at their highest in the two types of mares. These results indicate, a need for further studies on the effect of mare size or type on circulating concentrations of progesterone, and the consideration of size or type in clinically evaluating progesterone levels in pregnant mares. Apparent partial recovery of the corpus luteum of the previous estrous cycle occurred during early pregnancy in one pony.Increasing diameter of the embryonic vesicle and the extent of uterine tone were similar between horses and ponies. Although diameter of the uterine horns decreased in parallel for the two mare types during Days 0 to 40, the diameter was significantly less (2.2 mm difference averaged over all days) for ponies than horses. The temporal relationship between an expanding embryonic vesicle concurrently with decreasing uterine horn size and enhanced uterine turgidity is compatible with the postulated mechanism of embryo fixation. The smaller uterus in ponies with no difference in size of the conceptus between mare types could account for earlier fixation of the embryonic vesicle in ponies than in horses.     

Changes of receptor mRNAs for oxytocin and estrogen during the estrous cycle in rat uterus

Oxytocin receptor (OTR) mRNA levels increase dramatically near term and is potently stimulated by estrogen because increased OTR mRNA levels result from estrogen treatment in ovariectomized rat uterus. In this study, OTR, estrogen receptor (ER) alpha and ERbeta mRNA levels in the rat uterus during the estrous cycle were examined by quantitative RT-PCR. OTR mRNA levels during the estrous cycle began to increase on diestrus (P<0.05, vs value on estrus), reached maximal increase both in the morning (1000-1130 hr) and afternoon (1600-1630 hr) on proestrus (P<0.01, vs metestrus, diestrus and estrus) and then declined on estrus. In contrast ER alpha mRNA levels began to decrease on diestrus, reached statistical significance both in the morning and the afternoon on proestrus (P<0.01, vs metestrus, diestrus and estrus) and returned to the value of metestrus on estrus. ERbeta mRNA levels were low in the morning and the afternoon on proestrus (P<0.01, vs metestrus and estrus) and also returned to metestrus values on estrus. Treatments with estrogen for 3 days significantly decreased both ERalpha and ERbeta mRNA levels. It can be concluded from these results that during the estrous cycle, OTR mRNA levels in rat uterus predominantly increase at proestrus with a decrease in ERalpha and ERbeta mRNA levels, which is probably due to the increased estrogen levels in circulation before ovulation.     

Influence of bacterial challenge and long-term progesterone on endometritis of mares

Progress in treatment of endometritis in mares has been impeded by lack of an experimental model. This study tests the hypothesis that exposure of a mare's uterus to bacteria, while being treated with progesterone, will result in a compromised uterine defense after progesterone has been withdrawn. Ten mares were administered progesterone for 8 months and inoculated at the onset of treatment with a 5 x los colony forming units of(CFU) of Streptococcuszooepidemicus into the uterus. Mares were re-inoculated to maintain uterine infection, if necessary. Ten uninoculated mares with normal uteri served as controls. After the end of progesterone treat- ment all mares (treated and controls) experienced a normal estrous cycle and were inoculated with 5 x l0s CFU of Streptococcus zooepidemicus. Results of reproductive evalu- ations performed on days 2, 7 and 12 after bacterial inoculation were similar (P>0.05) for treated and controls. Pregnancy rates for treated and controls were also similar (P>0.05). Thus, the treatment evaluated in this study did not cause an alteration in the mare's ability to resolve endometritis.     

Lectin binding patterns of uterine glands in mares with chronic endometrial degeneration

OBJECTIVE: To evaluate changes of glycoconjugate in uterine glands of endometrial tissues obtained from mares. ANIMALS: adult mares. PROCEDURE: Uterine biopsy samples were collected during the breeding season and analyzed histologically for signs of chronic endometrial degeneration. Stage of the estrous cycle was established, using clinical examination and determination of hormonal status. Uterine tissue samples were analyzed, using lectin histochemical and immunohistochemical techniques (estrogen and progesterone receptors). Connective tissues were stained to determine alterations of ground substance in periglandular fibrosis. RESULTS: Of 50 mares, 30 (60%) were classified as normal or having modest alterations, and 20 (40%) were classified as having moderate or severe endometrial degeneration. In normal equine endometrium, several lectins (Helix pomatia agglutinin, Lotus tetragonolobus agglutinin, Ricinus communis I agglutinin, Ulex europaeus agglutinin, and wheat germ agglutinin) bound to glycoconjugates of the luminal epithelium and openings of uterine glands. Lectin binding patterns of cystic dilated glands or fibrotic glands in endometrial samples were remarkably strong, whereas normal surrounding cells remained unstained. Lotus tetragonolobus lectin was not suitable for detecting endometrial alterations. Connective tissues stained with Alcian blue and results of Hale colloidal-iron binding revealed acidic ground substance in periglandular fibrosis. Estrogen and progesterone receptors were evenly distributed in healthy and affected endometrial samples. CONCLUSIONS AND CLINICAL RELEVANCE: Glycoconjugate patterns of uterine glands were altered in mares with chronic endometrial degeneration. Therefore, uterine secretions are likely to be altered. These changes are not induced by changes in content of estrogen and progesterone receptors in endometrial tissues.     

Effects of Calf Removal at Parturition on Postpartum Ovarian Activity in Zebu (Bos indicus) Cows in the Humid Tropics

To assess endocrine and morphological responses of ovaries to total weaning at parturition, 6 Zebu (Bos indicus) cows 5 years or older were investigated. Following parturition, blood samples were collected daily during the first month and twice weekly thereafter until day 60 to determine concentrations of progesterone (P₄) and prostaglandin F_2α metabolite.It took between 25 to 32 days to complete uterine involution.The prostaglandin metabolite remained elevated for a mean period of 14.2 days (range, 4-21) postpartum. Five of the animals resumed cyclicity with a short estrous cycle starting between days 7 to 34 and lasting between 7 and 14 days. No estrous behavior was recorded prior to the short estrous cycles, but subsequent normal-length estrous cycles were all preceded by signs of estrus. In the 1 animal that resumed cyclicity with an estrous cycle of normal length on day 37 (length 20 days), the cycle was preceded by estrous behavior.Progesterone concentrations reached a mean maximum of 4.8 nmol liter⁻¹ during the short estrous cycles, and prostaglandin metabolite concentrations peaked while P₄ concentrations were decreasing. P₄ concentrations reached a mean maximum of 12.2 nmol liter⁻¹ during the estrous cycles of normal length. The interval from parturition to the first estrous cycle of normal length varied between 16 and 48 days, and the length of the cycle was 18 to 22 days.Starting 2 days postpartum, ovaries from 5 of the cows were scanned by ultrasonography every second day until day 30 postpartum. Medium-sized follicles were detected between days 4 to 7 postpartum in 4 of the scanned cows that later had short estrous cycles. The time between parturition and the appearance of the first dominant follicle was 7.6 days (range 6-10 days). The interval between parturition and the appearance of the first ovulatory-sized follicle was 10.2 days (range 8-13 days). In 3 of the scanned cows this ovulatory-sized follicle ovulated.We conclude that cyclic ovarian activity in Zebu cows can start early in the postpartum period in the absence of offspring, and that short luteal phases, not preceded by estrous behavior, may play an important role in establishing normal postpartum ovarian activity.     

Effect of Chronic Administration of Oxytocin on Corpus Luteum Function in Cycling Mares

The objective of this study was to determine if intramuscular administration of 60 units of oxytocin once daily for 29 days, regardless of when treatment was initiated during the estrous cycle (i.e., without monitoring estrous behavior and/or detecting ovulation), would induce prolonged corpus luteum (CL) function in cycling mares. Mares were randomly assigned to two groups: (1) saline-treated control (n = 7) and (2) oxytocin-treated (n = 9) subjects. Control mares received 3 cc of saline, and oxytocin-treated mares received 60 units (3 cc) of oxytocin intramuscularly for 29 consecutive days. Treatment was initiated in all mares on the same day (day 1), independent of the day of the cycle. Jugular blood samples for determination of progesterone concentration were collected three times weekly (M, W, and F) for 21 days before treatment was initiated to confirm that all mares had a luteal phase of normal duration immediately before treatment. Beginning on the first day of treatment, blood samples were collected daily for eight days and then three times weekly through day 80. Mares were considered to have prolonged CL function if serum progesterone remained >1.0 ng/mL continuously for at least 25 days after the end of the treatment period. The proportion of mares with prolonged CL function was higher in the oxytocin-treated group than in the saline-treated group (7/9 vs. 1/7, respectively; P < .05). Three of the seven oxytocin-treated mares that developed prolonged CL function initially underwent luteolysis within 4–7 days of the start of oxytocin treatment and then developed prolonged CL function after the subsequent ovulation during the treatment period. In the other four oxytocin-treated mares that developed prolonged CL function, progesterone remained >1.0 ng/mL throughout the treatment period and into the post-treatment period. All mares with prolonged CL function maintained elevated progesterone concentrations through at least day 55 of the study. In conclusion, intramuscular administration of 60 units of oxytocin for 29 consecutive days effectively prolonged CL function in mares, regardless of when treatment was initiated during the estrous cycle. Importantly, this represents a protocol for using oxytocin treatment to prolong CL function that does not require detection of estrous behavior or day of ovulation.     

Pregnancy diagnosis in thoroughbred mares using radioimmunoassay (RIA), enzyme immunoassay (kit) and ultrasound echography (scanning)

On the basis of progesterone determination on plasma or blood after RIA and the kit method respectively and consecutive scanning performed on a total number of 31 mares the following features were demonstrated: The overall material shows that in 20 mares (64.5%) embryonic vesicles were demonstrated. Of these mares 16 have conceived after service in the 1st estrous cycle and 3 mares in the 2nd estrous cycle. 18 mares were scanned in the time interval 13th-26th day after the latest service, while 2 mares were scanned on day 46 and 41 respectively. A total of 14 scanning positive mares were examined for progesterone by the RIA as well as by the kit method. For these mares (100%) agreement was found between the progesterone analyses as well as with the scanning results. For 19 mares (100%) there was agreement between kit and scanning results. 15 RIA progesterone determinations are in agreement with the scanning results. Progesterone values after the RIA method performed on 15 scanning positive mares were in average 8.40 +/- 2.79 ng/ml plasma. A total of 9 out of 10 scanning negative mares have been examined by the RIA as well as by the kit method. In 6 of these mares with 7 estrous cycles agreement has been found between progesterone analyses and the scanning results (77.8%). In 2 mares (no. 2 and 4) discrepancy has been found in the 2nd estrous cycle between RIA, kit and scanning results. For 1 mare (no. 7) discrepancy has been found in the 1st and 3rd estrous cycle between RIA and scanning.(ABSTRACT TRUNCATED AT 250 WORDS)     

Assessment of Uterine Vascular Perfusion During the Estrous Cycle of Mares in Connection to Circulating Leptin and Nitric Oxide Concentrations

The objective of this study was to find difference in vascular perfusion of uterine horns or uterine body throughout the estrous cycle and their relation to circulating nitric oxide and leptin concentrations. Five cyclic mares were subjected to transrectal Doppler ultrasonography and blood sampling for 18 days. Area of color and power Doppler modes was measured in pixels. Day (P = .0001) of the estrous cycle and ovulation (P = .0001) influenced uterine blood flow. Uterine body blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day 0 (day of ovulation), and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until days 12 and 10, respectively. Conversely to the contralateral uterine horn, ipsilateral uterine horn blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day −1, and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until day 10. Nitric oxide concentrations (P = .0001) attained two major peaks; the first on day −3 and the other persisted from day 2 until day 5. Leptin concentrations increased (P > .001) with a maximum value on day 0 and then decreased until a minimum value on day 9. In conclusion, during the estrous cycle, ipsilateral uterine horn and uterine body blood vessels had similar blood flow. Both leptin and nitric oxide played a role during follicle growth, ovulation, and corpus luteum development and modulated uterine blood flow before and after ovulation.     

Changes in oestrogen,progesterone and epidermal growth factor receptor concentrations and affinities during the oestrous cycle in the normal mammary gland and uterus of dogs

Changes in the concentrations and affinities of receptors for oestrogen (ER), progesterone (PR) and epidermal growth factor (EGF-R) were studied in mammary glands of healthy bitches with regard to age, the location in the mammary chain and the stage of the oestrous cycle. Uterus was used as the reference tissue for the evaluation of steroid receptors. Mammary and uterine samples from 7 healthy bitches were taken at five stages of the oestrous cycle in such a way that all the locations in the mammary chain were represented at each stage of the cycle (10 samples/dog). ER, PR and EGF-R were detected by biochemical assays using increasing concentrations of tritiated (steroids) or iodinated (EGF) ligands. A significant direct correlation was found between the ER and PR concentrations for mammary and uterine samples. No significant correlation was found between the steroid receptors and EGF-R concentrations. Mammary ER concentrations were significantly higher in bitches of 5 years of age or older than in younger ones; in posterior glands (4th and 5th pairs) than in anterior glands; and in the mid-luteal phase. Mammary PR did not vary significantly with age or location but was significantly lower in the early luteal phase than in other phases. A similar decrease in PR concentrations was observed in the uterus during the early luteal phase and uterine ER and PR concentrations were very low in the mid-luteal phase. Mammary EGF-R were not significantly higher in the early or mid-luteal phase than in pro-oestrus or anoestrus.The differences observed between the uterine and mammary steroid receptor concentrations during the oestrous cycle could be due to different mechanisms for regulating steroid receptor expression in the two tissues. Mammary EGF-R concentrations may be linked, as in other species, to cellular proliferation and/or to the serum progesterone concentrations.Abbreviations BSA bovine serum albumin - EGF epidermal growth factor - EGF-R receptor for epidermal growth factor - ER oestrogen receptor - K _d dissociation constant - LH luteinizing hormone - p probability of error - PR progesterone receptor