Effect of Dose of Cloprostenol on the Interval to Ovulation in the Diestrous Mare: A Retrospective Study

Although the ovulatory effects of prostaglandins are well documented in several domestic species including horses, there has been little attention paid to the use of this ovulatory effect for clinical purposes. Mares often grow large follicles during the luteal phase that may or may not ovulate before progesterone levels decline. Clinical observations of administering prostaglandins in diestrous mares with large follicles suggest that there may be a negative correlation between follicular diameter and interval from treatment to ovulation. The objectives of this study were twofold: to investigate the cloprostenol dose rate effect on interval to ovulation and to confirm the negative correlation between follicular diameter and interval to ovulation. The hypothesis tested was that high doses of cloprostenol given in diestrus to mares with larger follicles would induce ovulation more rapidly than in mares given lower doses or with smaller follicles. To test the hypothesis, a total of 1,234 estrous cycles were induced with different doses of cloprostenol (ranging from 8.75 to 625 μg). All mares had at least one follicle of 28 mm or larger. Dominant follicles were followed by transrectal ultrasound examinations every other day until ovulation was detected. There was a significant effect of dose (P < .000) and follicular diameter (P < .000) on the interval from treatment to ovulation. The shortest mean interval (2.4 days) was observed after administration of 625 μg in mares with follicles 36 mm or larger, whereas the longest (4.9 days) occurred after 8.75 μg in follicles of 28 to 31 mm.     

Incidence, Endocrinology, Vascularity, and Morphology of Hemorrhagic Anovulatory Follicles in Mares

The incidence of hemorrhagic anovulatory follicles (HAFs) is approximately 5% and 20% of estrous cycles during the early and late ovulatory season, respectively. The structures are more common in old mares (eg, >20 years), tend to occur repeatedly in individuals, and occur most frequently during the late follicular phase. In a recent study, the day of ovulation in controls and the first day of HAF formation, as indicated by cloudiness of follicular fluid, were defined as day 0. On day -1, future ovulating and HAF groups did not differ in follicle diameter or in the frequency of discrete gray-scale ultrasonic indicators of impending ovulation; however, in future HAFs, a greater percentage of the circumference of the follicle exhibited color-Doppler signals of blood flow. No differences were found between the two groups in systemic concentrations of progesterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) on days -4 to 2, but estradiol was elevated in the HAF group on day -3. The wall of the HAFs developed well-vascularized luteal tissue as indicated by echotexture and color Doppler signals and by the production of near normal levels of progesterone. In conclusion, HAFs formed from viable preovulatory follicles that did not differ from ovulatory follicles in diameter or gray-scale echotexture. Estradiol concentrations were elevated a few days before the failure of ovulation, and the wall of the follicle was more extensively vascularized on day -1.     

Expression of messenger ribonucleic acid encoding for steroidogenic acute regulatory protein and enzymes, and luteinizing hormone receptor during the spring transitional season in equine follicles

The period of spring transition, from the anovulatory to the ovulatory season, is characterized in many mares by cyclical growth and regression of large dominant follicles. These follicles produce only low concentrations of estradiol and it is thought that acquisition of steroidogenic competence by large follicles during spring transition is prerequisite in stimulating LH prior to first ovulation. In situ hybridization was used to localize and quantify expression of factors that play a key role in follicular steroidogenesis: StAR, P450scc (CYP11A1), P450c17 (CYP17), P450arom (CYP19), and LH receptor (LHr). One ovary was obtained from mares on the day after detection of an actively growing 30 mm transitional anovulatory follicle (defined as the transitional follicle), and the remaining ovary was removed at the third estrus of the breeding season on the day after the preovulatory follicle reached 30 mm in diameter (defined as the preovulatory follicle). Messenger RNAs encoding StAR, CYP11A1, and CYP17 were detected only in theca cells and CYP19 mRNA was confined to the granulosa layer. There was significantly lower expression of mRNAs for the steroidogenic enzymes, StAR (P<0.001) and LHr (P<0.05) in transitional follicles than in preovulatory follicles. In conclusion, large equine follicles during spring transition have low levels of mRNA encoding steroidogenic enzymes, StAR and LHr which will contribute to the steroidogenic incompetence of dominant follicles during spring transition and their subsequent regression.     

Ovarian follicular dynamics in heifers: test of two-wave hypothesis by ultrasonically monitoring individual follicles

The two-wave hypothesis for follicular development during the bovine estrous cycle was tested by ultrasonically monitoring individual follicles in 10 heifers during an interovulatory interval. A dominant follicle was defined as one that reached a diameter of at least 11 mm. Subordinate follicles were defined as those that appeared to originate from the same follicular pool as a dominant follicle. A dominant follicle and its cohorts were defined as a wave. Two waves during an interovulatory interval were identified in 9 of 10 heifers. The first wave was first identified, retrospectively, on a mean of Day 0.2 +/- 0.1 (ovulation = Day 0) and gave origin to a dominant anovulatory follicle and a mean of 1.4 +/- 0.3 identified subordinates. The dominant follicle reached maximum diameter (mean, 15.8 +/- 0.8 mm) on an average of Day 7 and then decreased (P less than .04) by Day 11. The subordinate follicles increased in diameter for a few days and then regressed. The second wave was first identified on a mean of Day 10.0 +/- 0.4 and gave origin to the ovulatory follicle and a mean of 0.9 +/- 0.3 subordinates. One of the 10 heifers had 3 waves of follicular activity characterized by an anovulatory wave emerging on Day 0, another anovulatory wave emerging on Day 10, and an ovulatory wave emerging on Day 16. Results strongly supported the two-wave hypothesis but also indicated that a minority of interovulatory intervals in this heifer population may have 3 waves of follicular activity.     

Follicle Development in Mares

The mare provides a unique experimental model for studying follicle development in monovular species. Development of antral follicles in horses is characterized by the periodic growth of follicular waves which often involve the selection of a single dominant follicle. If properly stimulated, the dominant follicle will complete development and eventually ovulate a fertile oocyte. Regulation of follicular wave emergence and follicle selection involves an interplay between circulating gonadotropins and follicular factors that ensures that individual follicles are properly stimulated to grow (or to regress) at any given stage of follicular wave development. Periodic development of follicular waves continuously occurs during most of post-natal life in the mare and is influenced by factors such as stage of oestrous cycle, season, pregnancy, age, breed and individual so that different types of follicular waves (minor or major, ovulatory or anovulatory) and different levels of activity within waves may develop under different physiological conditions. Changes in gonadotropin levels and/or in the sensitivity of follicles to circulating gonadotropins seem to account largely for these physiological variations in follicle development.     

Follicular fluid concentrations of free insulin-like growth factor (IGF)-I during follicular development in mares

The objective of the present study was to evaluate changes in concentrations of free insulin-like growth factor (IGF)-I in follicular fluid (FFL) during follicle development in the mare. Mares (n = 14) were classified as either in the follicular phase (n = 8) or luteal phase (n = 6). Follicles (n = 92) were categorized as small (6–15 mm; n = 54), medium (16–25 mm; n = 23) or large (>25 mm; n = 15) and FFL was collected. Free IGF-I levels in FFL in large follicles of follicular phase mares were greater (P < 0.05) than in large follicles of luteal phase mares and small or medium follicles of luteal and follicular phase mares. Free IGF-I concentrations were greater (P < 0.05) in large follicles of luteal phase mares than small but not medium follicles of luteal phase mares. FFL ratio of estradiol:progesterone paralleled changes in free IGF-I. Free IGF-I concentrations were negatively correlated (P < 0.05) with insulin-like growth factor binding protein (IGFBP)-2, -4 and -5 but not IGFBP-3 levels. In addition, free IGF-I concentrations in FFL were positively correlated (P < 0.01) with FFL estradiol, progesterone, androstenedione, estradiol:progesterone ratio, total IGF-I and total IGF-II. We conclude that increases in intrafollicular levels of bioavailable (free) IGF-I are associated with increased steroidogenesis in developing mare follicles.     

Selected Ovarian Ultrasonographic Characteristics During Vernal Transition are Useful to Estimate Time of First Ovulation of the Year

It is important to get mares pregnant as early as possible after vernal transition and thus, identification signs of impending 1st ovulation of the year are warranted. To identify clinical indicators of an approaching first ovulation of the year, mares were teased with a stallion for oestrous detection starting January 3 and subjected to ultrasonographic examination. Day of first appearance of uterus oedema, follicular wall invagination, intrafollicular echogenicity, double contour of the follicle wall, increase in granulosa thickness, follicular wall hyperechogenicity and appearance of pear‐shaped follicles was registered, as well as follicle diameter and number. Seventy per cent of the mares had anovulatory oestrous periods of 4.6 ± 3.6 days, with an interoestroual interval of 12.5 ± 12.2 days. Number of anovulatory oestruses per mare was 2.4 ± 2.3. Uterine oedema occurred in 77% of the mares, 32.4 ± 25.6 days before ovulation. Invagination of the follicular wall appeared in 44.4% of the animals, 24.5 ± 18.4 days before ovulation. Intrafollicular echogenicity was seen in all mares and double contour of the follicle was seen in 77% of the animals. Both last two characteristics appeared 1–72 days before ovulation. Increased thickness of the granulosa occurred in 66% of the mares, 1–19 days before ovulation. Pear‐shaped follicles and follicular wall hyperechogenicity were detected 3 or less days before the first ovulation, in 44.4% and 55.5% of mares, respectively. Mean number of follicles >15 mm decreased at least 16 days before ovulation. We concluded that no isolated characteristic was a reliable indicator. However, increase in granulosa thickness, formation of a pear‐shaped follicle and follicular wall hyperechogenicity, associated with the reduction of the number of follicles >15 mm in diameter to <3, resulted in the first ovulation of the year in 44–67% of the transitional mares, 1–19 days after the characteristics appeared.     

Changes in follicular fluid steroids, insulin-like growth factors (IGF) and IGF-binding protein concentration, and proteolytic activity during equine follicular development

The objective of the present study was to evaluate changes in equine follicular fluid insulin-like growth factor binding protein (IGFBP) proteolytic activity as well as steroid, IGF, and IGFBP concentrations during follicular development in the mare. Mares (n = 14) were classified as either in the follicular phase (n = 8) or luteal phase (n = 6). Follicles (n = 92) were categorized as small (6 to 15 mm; n = 54), medium (16 to 25 mm; n = 23), or large (> 25 mm; n = 15), and follicular fluid was collected. Estradiol and androstenedione levels in follicular fluid were greater (P < 0.05), and IGFBP-3 concentrations tended to be greater (P < 0.10) in large than in small or medium follicles, whereas IGFBP-2, -4, and -5 levels were less (P < 0.05) in large than in small or medium follicles. Estradiol and androstenedione concentrations were negatively correlated (P < 0.01) with IGFBP-2, -4, and -5 but not IGFBP-3 concentrations. To evaluate proteolysis of IGFBP, follicular fluid was incubated with human 125I-labeled IGFBP-2, -3, and -5 and protein separated by 12% SDS-PAGE. Follicular fluid caused little or no proteolysis of 125I-lableled IGFBP-2 or -3, and the small amount of proteolysis of IGFBP-2 and -3 did not differ (P > 0.10) among follicle classes. However, more 125I-labeled IGFBP-5 was cleaved (P < 0.05) by follicular fluid from large follicles collected during the follicular phase than large follicles during the luteal phase, and small or medium follicles from follicular and luteal phase mares indicating that a protease to IGFBP-5 exists in estrogen-dominant equine follicles. This IGFBP-5 protease was inhibited by kallikrein/serine protease and metalloprotease inhibitors. We conclude that the tendency of estrogen-dominant follicles of mares to have greater levels of IGFBP-3 and lesser levels of IGFBP-2 does not appear to be due to differences in proteolysis, whereas changes in IGFBP-5 levels are likely due to changes in activity of a serine protease or metalloprotease. Changes in IGFBP may alter levels of bioavailable IGF that stimulate steroidogenesis and mitogenesis in developing mare follicles.     

Comparison of Different Treatments for Oestrous Induction in Seasonally Anovulatory Mares

The aim of this study was to evaluate the effects of different treatments for induction and synchronization of oestrus and ovulation in seasonally anovulatory mares. Fifteen mares formed the control group (C), while 26 mares were randomly assigned to three treatment groups. Group T1 (n = 11) were treated with oral altrenogest (0.044 mg/kg; Regumate^®) during 11 days. Group T2 (n = 7) was intravaginally treated with 1.38 g of progesterone (CIDR^®) for 11 days. In group T3 (n = 8), mares were also treated with CIDR^®, but only for 8 days. All mares received PGF2α 1 day after finishing the treatment. Sonographic evaluation of follicles, pre‐ovulatory follicle size and ovulation time was recorded. Progesterone and leptin levels were analysed. Results show that pre‐ovulatory follicles were developed after the treatment in 88.5% of mares. However, the pre‐ovulatory follicle growth was dispersal, and sometimes it was detected when treatment was not finished. While in mares treated with intravaginal device, the follicle was soon detected (1.5 ± 1.2 days and 2.3 ± 2.0 days in T2 and T3 groups, respectively), in T1 group, the pre‐ovulatory follicle was detected slightly later (3.9 ± 1.6 days). The interval from the end of treatment to ovulation did not show significant differences between groups (T1 = 13.1 ± 2.5 days; T2 = 11.0 ± 3.6 days; T3 = 13.8 ± 4.3 days). The pregnancy rate was 47.4%, similar to the rate observed in group C (46.7%; p > 0.05). Initial leptin concentrations were significantly higher in mares, which restart their ovarian activity after treatments, suggesting a role in the reproduction mechanisms in mares. It could be concluded that the used treatments may be effective for oestrous induction in mares during the late phase of the seasonally anovulatory period. Furthermore, they cannot synchronize oestrus, and then, it is necessary to know the reproductive status of mares when these treatments are used for oestrous synchronization.     

In vivo evidence that local cortisol production increases in the preovulatory follicle of the cow

The aim of the present in vivo study was to monitor real-time fluctuations of cortisol (Cr) in the wall of preovulatory follicles using a microdialysis system (MDS) implanted in the theca layer as well as changes in ovarian venous plasma (OVP) and jugular venous plasma (JVP). Seven cows were superovulated using FSH and prostaglandin F2alpha injections. Dialysis capillary membranes were surgically implanted into the theca layer of mature follicles and connected to a microdialysis system. Fractions of the perfusates were collected from Day -1 (Day 0=LH surge) to Day 3. No difference in the concentrations of Cr between JVP and OVP was detected throughout the experiment. Circulating concentrations of Cr ranged from 20 to 35 ng/ml 8 h after surgery in ovulatory and anovulatory cows. In five ovulatory cows, the Cr concentration decreased to basal levels (<10 ng/ml) between 12 and 24 h after surgery, however, two anovulatory cows retained high Cr levels (>10 ng/ml) up to 42 h after surgery. There was a clear increase in the local concentration of Cr from 13.3+/-2.1 pg/ml at -24 h to 27.5+/-1.7 pg/ml at 0 h (peak of the LH surge) within the wall of ovulatory follicles. This increase was not detected in anovulatory follicles. This transient increase in Cr occurred only in the follicle wall, but not in the OVP or JVP, indicating that the presence of a local regulatory mechanism for Cr production/conversion in ovulatory follicles, and this mechanism may modulate the inflammatory-like reaction induced by LH surge in the follicle wall. The present results demonstrate that the glucocorticoid environment in the follicular wall adjusts at the local level in bovine ovulatory follicles. This mechanism may protect follicles from the adverse effects of glucocorticoid, and it may prevent excess inflammatory reactions associated with ovulation by temporarily increasing local concentrations of glucocorticoid, thus forming an integral part of the regulatory mechanism in ovarian physiology.     

Ultrasound Confirmation of Ovulation in Mares: A Normal Corpus Luteum or a Haemorrhagic Anovulatory Follicle?

The most common pathological anovulatory condition that occurs spontaneously during the breeding season in the mare is the haemorrhagic anovulatory follicle (HAF). A relatively high proportion of mares, soon after ovulation, develop a corpus haemorrhagicum (CH) with a central lacuna. This type of corpora lutea may resemble an HAF, which may complicate the accurate diagnosis of ovulation. The main objective of this study was to compare the ultrasound data of mares examined frequently with HAFs and CHs to elucidate whether it is possible to distinguish them from each other. A total of 135 ovulating mares were classified according to the morphology of the corpus luteum (CL) in mares with: a solid CL, a CH with small or with large central cavities. Ultrasound characteristics of the development of 11 HAF and 13 CHs with a large central cavity were compared. The pre‐ovulatory follicular diameter of ovulatory mares was significantly correlated with the diameter of CH with large central cavities. The percentage of mares with post‐ovulatory areas eligible to be mistaken with a CH was <25%. Although a predictive diagnosis of an HAF/CH can be made on the basis of several ultrasonographic endpoints, the only parameter that allows a definitive diagnosis is the thickness of the luteal border. This is <3 mm in HAFs in contrast to >5 mm in CHs. However, this only applies when the unidentified structure has non‐organized contents.     

Ovarian activity of the mare during winter and spring at a latitude of 19° 21′ north

The ovulatory activity of mares located at 19° 21′N was monitored from November to June. Twenty-one mares were used (11 quarter-mile mares and 10 hybrids). Every week the mares were subjected to ultrasonographic observations of the ovaries, and blood samples for progesterone determinations were collected. Estrus activity was evaluated by teasing with a stallion. A mare was considered to be in an anovulatory state if progesterone concentrations during 5 or more consecutive samples were below 1 ng/mL. Cyclic ovulatory activity during the entire duration of the study was recorded in 62% of the mares. Only 38% of the mares (5 quarter-mile and 3 hybrid mares) had anovulatory periods, which lasted 128.5 ± 14.4 days (mean ± SEM) and occurred at some point between November and May. The duration of the anovulatory period varied from 70 to 196 days. Estrous behavior was shown during the anovulatory period in 75% (6/8) of the mares that had such a period. Most mares maintain regular ovulatory activity during the winter and spring at 19° 21′ north, and that a wide variation occurs in the duration of the anovulatory period of those mares that stop cycling at this latitude.     

Does Clinical Treatment with Phenylbutazone and Meloxicam in the Pre‐ovulatory Period Influence the Ovulation Rate in Mares?

The presence of anovulatory haemorrhagic follicles during the oestrous cycle of mares causes financial impacts, slowing conception and increasing the number of services per pregnancy. Non‐steroidal anti‐inflammatory drugs (NSAIDs) such as meloxicam and phenylbutazone are used in the treatment of several disorders in mares, and these drugs can impair the formation of prostaglandins (PGs) and consequently interfere with reproductive activity. This study aimed to evaluate the effects of treatment with NSAIDs on the development of pre‐ovulatory follicles in mares. In total, 11 mares were studied over three consecutive oestrous cycles, and gynaecological and ultrasound examinations were performed every 12 h. When 32‐mm‐diameter follicles were detected, 1 mg of deslorelin was administered to induce ovulation. The first cycle was used as a control, and the mares received only a dose of deslorelin. In the subsequent cycles, in addition to receiving the same dose of deslorelin, each mare was treated with NSAIDs. In the second cycle, 4.4 mg/kg of phenylbutazone was administered, and in the third cycle, 0.6 mg/kg of meloxicam was administered once a day until ovulation or the beginning of follicular haemorrhage. All of the mares ovulated between 36 and 48 h after the induction in the control cycle. In the meloxicam cycle, 10 mares (92%) did not ovulate, while in the phenylbutazone cycle, nine mares (83%) did not ovulate. In both treatments, intrafollicular hyperechoic spots indicative of haemorrhagic follicles were observed on ultrasound. Thus, our results suggested that treatment with meloxicam and phenylbutazone at therapeutic doses induced intrafollicular haemorrhage and luteinization of anovulatory follicles.     

Induction of ovulation and multiple ovulations in seasonally anovulatory and ovulatory mares with an equine pituitary extract

A crude equine pituitary ethanol extract (EE) was used to induce single and miltiple ovulations in seasonally anovulatory pony mares 3-15 years of age. 12 mares were injected daily for 14 days with EE; 6 of the EE-treated mares were also treated with human chorionic gonadotropin (HCG), and 6 control mares received saline vehicle only. In a 2nd experiment designed to determine if EE treatment could induce multiple ovulations in seasonally ovulatory mares, 7 mares were treated during diestrus, 7 mares were treated beginning on Day 1 of estrus, and 7 remained untreated. The results of experiment 1 confirmed that EE treatment can induce ovulation in mares during the anovulatory season, that the timing of ovulation can be improved with HCG, and that ova from induced ovulations are fertilizable. Results of experiment 2 demonstrated that EE treatment can induce follicular activity and multiple ovulations during the ovulatory season.     

Characterization of immunoreactive IGF-I pattern during the peri-ovulatory period of the oestrous cycle of thoroughbred mares and its relation to other hormones

The aim of this study was to characterize ir-IGF-I pattern and its relation to other hormones during the oestrous cycle in mares. Nine non-pregnant non-lactating pluriparous thoroughbred mares were used. The studied mares were examined ultrasonically and bled daily to follow the ovarian changes and the hormonal milieu for a complete Interovulatory interval (IOI). Two (minor and major) follicular waves were characterized per IOI in thoroughbred mares. The largest follicle of the first follicular wave (DF1) was firstly detected at D - 1.75 ± 0.47 with a growth rate of 2.78 ± 0.14mm/day and maximum diameter of 22.45 ± 0.75mm on day 6.65 ± 0.82. The largest follicle of the second follicular wave (DF2) had a growth rate of 2.15 ± 0.29 mm/day, reached a maximum diameter of 42.70 ± 2.63 mm on D 19.25 ± 0.43. Ir-IGF-I increased significantly prior to ovulation and had a similar pattern to oestrogen (r = 0.84, p < 0.05), suggesting that the ovarian follicles are the main source of circulating ir-IGF-I during the oestrous cycle of mares and that ir-IGF-I may be a crucial factor in follicular differentiation and maturation. In conclusion, this study demonstrated that ir-IGF-I is secreted during the oestrous phase of the cycle concomitant with the development of the future ovulatory dominant follicle, and it may act in synergy with other hormones for the selection and differentiation of the dominant follicle.     

Risk Factors for the Development of Haemorrhagic Anovulatory Follicles in the Mare

Haemorrhage into the dominant follicle during the reproductive season is a subtle but definitive cause of infertility in the mare population. This condition however can be of high relevance for an individual in which its incidence is abnormally high. Little is known about the nature and factors affecting the incidence of haemorrhagic anovulatory follicles (HAFs) in the mare. The objectives of the study were to define and characterize the ultrasonographic development and incidence of HAFs and to investigate possible risk factors influencing its occurrence. Detailed reproductive and ultrasound records of seven mares studied during their entire reproductive lives (>10 years and 612 oestrous cycles) were analysed retrospectively and computed into a statistical mixed model. Of all animal studied, two mares were found to have an unusually high incidence of HAFs of approximately 25%. Time of season and use of induction treatments (Cloprostenol) were found to influence its incidence. It appears that early‐enhanced stimulatory effect of LH on an ovary with the presence of small and immature follicles might increase the risk of ovulatory failure of those follicles later in the cycle. Mares during the months of highest follicular activity (May to August) and after treatment with hormones to induce oestrus and ovulation are at greater risk to develop HAFs. The potential relevance of this study is two folds: clinical relevance for the practitioner to better understand this condition and so improve reproductive management of mares with abnormally high incidence; and to provide useful insights for researchers willing to further investigate the nature of this phenomenon.     

Ultrasonic evaluation of the reproductive tract of the mare: Ovaries

Ultrasonography is useful for monitoring the dynamic follicular and luteal changes of equine ovaries, since it permits rapid, visual, non-invasive access to the reproductive tract. A 5 MHz transducer has greater resolving power and is far more suitable for evaluation of ovaries than a 3–3.5 MHz transducer. Follicles as small as 2–3 mm can be seen and the corpus luteum can usually be identified throughout its functional life. In a study involving daily ovarian examinations, there was a pronounced change in shape of the preovulatory follicle from a roughly spherical to a pear-shaped or oblong form in 66% of the ovulatory periods, This change usually occurred on the day preceding ovulation. The occurrence of ovulation was detectable by the disappearance of a large follicle. In addition, the ovulation site on day 0 was characterized by an intense echogenic area in 88% of 32 ovulations. The developing corpus luteum retained the echogenicity for a mean of 2.4 days. In a blind study, the location of the corpus luteum, as determined by ultrasound, agreed with a previous independent determination of the side of ovulation by palpation in 88% of the 40 bred mares on days 0–14. In all of the 12 mares that were in estrus, the location of the corpus luteum could not be ascertained. In another study, the corpus luteum was identified for a mean of 16 days in 14 estrous cycles. One or more days before the corpus luteum became ultrasonically unidentifiable, it developed increased echogenicity in 36% of the mares, indicating greater tissue density. It is concluded that ultrasonic evaluation of the corpus luteum is superior to digital evaluation by rectal palpation. Some of the potential applications of ultrasonic examination of the ovaries include: 1) obtaining important, sometimes definitive, information by a single examination for judging whether a mare has entered the ovulatory season, 2) aiding in estimating the stage of the estrous cycle, 3) detecting double preovulatory-sized follicles which are in close apposition and difficult to discern by palpation, 4) detecting failure of ovulation or anovulatory estrus by the absence of a corpus luteum, 5) differentiating a persistent corpus luteum from anovulatory or anestrous conditions, 6) diagnosing certain pathological conditions such as peri-ovarian cysts and ovarian tumors, and 7) diagnosing anovulatory hemorrhagic follicles.     

Increased Frequency of Double Ovulations after Induction of Luteolysis with Exogenous Prostaglandin F2α

The effect of induction of luteolysis by intramuscular treatment with prostaglandin F2α (PGF) on the frequency of double ovulations and formation of hemorrhagic anovulatory follicles (HAFs) was studied. The PGF (5 mg) was given 10 days after ovulation (n = 47 estrous cycles). No treatment or sham injection was used for control estrous cycles (n = 39). After treatment, the mares were scanned by transrectal ultrasonic imaging every 2 days until the largest follicle reached 25 mm and every day thereafter until the outcome of all follicles of at least 25 mm was determined. The frequency of two ovulations during the posttreatment ovulatory period was greater (P < .03) in the treated group (17%) than in the controls (3%). The combined frequency of two ovulations or one ovulation and one HAF also was greater (P < .002) in the treated group (30% vs. 5%). Equine veterinarians should be aware that PGF induction of luteolysis may increase the frequency of double ovulations or HAFs.     

Metabolic profiles in ovulatory and anovulatory primiparous dairy cows during the first follicular wave postpartum

Metabolic hormones affect ovarian function in the cow. However, the relationship between metabolic factors and ovarian function is not clear in the postpartum primiparous cow because they are still growing. The aim of the present study was to investigate in detail the time-dependent profile of the metabolic hormones, metabolites, and milk yields of ovulatory and anovulatory primiparous cows during the first follicular wave postpartum. We used 16 primiparous Holstein cows and obtained blood samples for the profiles of metabolites (glucose; non-esterified fatty acid, NEFA; ketone body; total cholesterol; and aspartate aminotransferase), metabolic hormones (growth hormone, GH; insulin-like growth factor-I, IGF-1; and insulin), and progesterone every other day from 1 to 21 days postpartum. In addition, all ovaries were observed using ultrasound. Dairy milk yield was recorded during the experimental period. In all cows, the first follicular wave postpartum was observed and 6 of the cows ovulated. The plasma glucose (P<0.0001) and IGF-1 (P<0.001) concentrations were lower and the plasma NEFA (P<0.0001) and ketone bodies (P<0.0001) concentrations and daily milk yield (P<0.0001) were higher in the anovulatory cows compared to the ovulatory cows. However, the GH levels, which enhance lipolysis for milk production, insulin and other metabolites did not differ between the two groups. In conclusion, the present study suggests that anovulation of the dominant follicle during the first follicular wave postpartum in primiparous cows is induced by low IGF-1 levels that are similar to those of multiparous cows. In addition, anovulatory cows are likely to mobilize body fat stores for milk production more easily than ovulatory cows.     

Insulin-like growth factor I in sera, ovarian follicles and follicular fluid of cows with spontaneous or induced cystic ovarian disease

The objective of this research was to determine changes in IGF-I levels in serum and follicular fluid, and immunoreactivity of the follicle wall of cows with spontaneous (slaughter specimens) or ACTH-induced follicular cysts, and to compare results to normal cycling (control) cows after selection of the ovulatory follicle. Concentrations of IGF-I in serum did not differ between control and cystic animals (p=0.76). Fluid from the ovulatory follicle in control cows had 41% higher concentrations of IGF-I than that from cystic follicles collected at slaughter (spontaneous cysts; p<0.05) and 70% higher than that in induced follicular cysts (p<0.05). An intense positive immunostaining with anti-IGF-I was observed in granulosa cells (p<0.05) and in the theca interna (p<0.05) of secondary and tertiary follicles in all three groups of animals, but staining was less intense in cystic (p<0.05) and atretic follicles (p<0.05). This study provides evidence to suggest that cystic ovarian disease in cattle is associated with decreased concentrations of IGF-I in follicular fluid, but not in serum, and decreased production of IGF-I in the follicular wall. These data support the notion that IGF-I plays a role in the regulation of folliculogenesis, and may participate in the pathogenesis of cystic ovarian disease in cattle.