The effects of an advanced uterine environment on embryonic survival in the mare

Reasons for performing the study: During embryo transfer (ET) the equine embryo can tolerate a wide degree of negative asynchrony but positive asynchrony of >2 days usually results in embryonic death. There is still confusion over whether this is due to the inability of the embryo to induce luteostasis or to an inappropriate uterine environment. Objectives: To assess embryo survival and development in an advanced uterine environment. Hypothesis: Embryo–uterine asynchrony, not the embryo's inability to induce luteostasis, is responsible for embryonic death in recipient mares with a >2 days chronologically advanced uterus. Methods: Experiment 1: Thirteen Day 7 embryos were transferred to the uteri of recipient mares with luteal prolongation, occasioned by manual crushing of their own conceptus, such that donor–recipient asynchrony was between +13 and +49 days. Experiment 2: Day 7 embryos were transferred to recipient mares carrying their own conceptus at Days 18 (n = 2), 15 (n = 2), 14 (n = 4), 12 (n = 4) or 11 (n = 4) of gestation. In addition, Day 8 embryos were transferred to 4 pregnant recipient mares on Day 11 of gestation. Results: No pregnancies resulted following transfer of Day 7 embryos to recipients in prolonged dioestrus with asynchronies between +13 and +49 days. However, the use of early pregnant mares as recipients resulted in 5/20 (25%) twin pregnancies, 4 of which came from the transfer of a Day 8 embryo to a Day 11 recipient. All transferred embryos showed retarded growth, with death occurring in 4/5 (80%). Conclusions and potential relevance: The results emphasise the importance of an appropriate uterine environment for embryo growth and the inability of equine embryos to survive transfer to a uterus >2 days advanced even when luteostasis is achieved. It is possible that in normal, non‐ET equine pregnancy, embryo–uterine asynchrony may account for some cases of embryonic death.     

Evaluation of progesterone treatment to create a model for equine endometritis.

To investigate a model for equine endometritis, 12 mares with normal reproductive tracts were divided into 2 groups. All mares received progesterone in oil, 250 mg im, daily. At 5 days after initiation of progesterone administration, the uteri were inoculated with 10(6) colony forming units of Pseudomonas aeruginosa. The day of inoculation was designated Day 0. On Day 6, endometrial swab samples yielded P. aeruginosa in 5 mares; samples from the other 7 mares yielded heavy growth of Escherichia coli, Streptococcus zooepidemicus, Klebsiella pneumoniae, Enterobacter spp., Citrobacter diversus, Staphylococcus epidermidis and Streptococcus morbillorum. On Days 6, 7 and 8, Group A mares received intrauterine infusions of 6 g ticarcillin disodium and 0.2 g clavulanate potassium in 100 ml sterile saline. Group B mares received infusions of saline only. The incidence of swab specimens yielding no bacterial growth was significantly higher in Group A than Group B mares on Days 8 and 13 (4/6 vs 0/6). Swab samples from 5 of the 6 mares in Group A yielded growth of fungi on Days 13 and 19. Mares in Group B were then similarly treated with ticarcillin/clavulanate infusions, on Days 19, 20 and 21. The incidence of swab specimens yielding no bacterial growth was 2/6 and 1/6 on Days 21 and 26, respectively; fungi were not recovered from these mares at any time. The incidence of no-growth swabs after antibiotic treatment tended to be higher in Group A and incidence of fungal recovery after antibiotic treatment was significantly higher in Group A.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in plasma progesterone concentrations from days 17 to 42 of gestation in mares maintaining or losing pregnancy

Plasma progesterone concentrations were measured in 179 mares bled on alternate days commencing with a positive pregnancy diagnosis on Days 17 to 18 after ovulation and concluding on Days 42 to 45. During this period 17 mares (10 per cent) lost their pregnancies, 11 before Day 25. In 15 mares the timing of the pregnancy loss could be determined with adequate accuracy; in only one did a decline in progesterone precede the loss. Thus pregnancy loss between Days 17 and 42 was rarely caused by a fall in plasma progesterone.     

Concentrations of uterine luminal prostaglandins in mares with acute and persistent endometritis

Intrauterine infusion of 1 per cent oyster glycogen solution was used to induce acute endometritis in four genitally normal mares. Numbers of viable neutrophils recovered in uterine washings had increased by 1 h after infusion and remained elevated for at least 72 h. There was a significant correlation between numbers of viable neutrophils and total protein concentrations and between prostaglandin (PG)F and PGE2 concentrations in washings. There was also a significant relationship between concentrations of 15-keto-13, 14-dihydro PGF2 alpha in plasma and PGF in washings. Intrauterine concentrations of PGF were influenced by cycle stage and in turn the induced acute endometritis interfered with normal ovarian function. Mares with persistent endometritis had significantly higher concentrations of PGF and total protein and percentage of neutrophils and mononuclear cells in washings than normal mares. White blood cells from mares were capable of producing PGF and PGE2 in vitro.     

Diagnosis and correction of twin pregnancy in the mare

Reproductive Physiology 1. Twin pregnancies result in high rates of abortion, stillbirth, and neonatal mortality. 2. Twins develop subsequent to multiple ovulations. Multiple ovulations are related to breed, parity, and mare history. Multiple ovulations are most frequently seen in Thoroughbred and Draft mares. Multiple ovulations are more common in barren and perhaps maiden mares than in lactating mares, and they are more common in certain individual mares. 3. Equine embryos are motile in the uterus from the time of first detection (Days 9 to 10) until fixation (Day 16). They are frequently located in the uterine body on Days 9 and 10. 4. Twin embryos have a pattern of motility and fixation similar to that of single embryos, and fixation is more frequently unilateral than bilateral (70 per cent versus 30 per cent, respectively). 5. Mares have an efficient natural embryo-reduction mechanism to eliminate excess (greater than 1) embryos resulting from multiple ovulations. Natural embryo reduction is more successful in unilateral than bilateral twin pregnancies (89 per cent versus 11 per cent successful reduction, respectively). 6. After the establishment of endometrial cups (Days 35 to 40), mares that are aborted will frequently not cycle for several months. Management of Twin Pregnancy 1. Breed all mares regardless of the number of preovulatory follicles. Withholding mares with preovulatory follicles from breeding does not decrease the incidence of twin pregnancy, but it decreases the overall pregnancy rate and results in a loss of breeding time. 2. Check all mares for twins, regardless of the number of detected ovulations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Exercising the Pregnant Mare from Day 16 to Day 80 of Gestation

Our previous research has demonstrated moderate exercise can be detrimental to early pregnancy in the mare, but little work has examined exercise after pregnancy has been detected. We exercised mares (n = 8) 6 days a week for 45 min from Day 16 until Day 80 of gestation. Color Doppler ultrasonography was used to evaluate embryonic vesicle size, fetal length, and uterine blood flow. Blood was sampled every other day to analyze cortisol and progesterone concentrations. Results indicated that exercising pregnant mares (n = 4) led to greater (P < .01) cortisol concentrations 30 min after the exercise period. No overall treatment effect could be detected in progesterone concentrations; however, following Day 60 of gestation, progesterone concentrations were lower (P < .05) in exercised mares. Additionally, progesterone concentrations peaked earlier in exercised mares at Day 52 of gestation compared to peak levels at Day 68 of gestation for control mares. No significant effects were detected in embryonic vesicle size. Fetal length tended (P = .06) to be longer in the conceptus of exercised mares. Uterine blood flow did not differ between groups but increased as pregnancy progressed (P < .001) in both groups. All mares in this study went on to deliver healthy foals and suffered no difficulties during parturition. These results indicated that moderate exercise was not detrimental to mare pregnancy.     

The technique and clinical application of endometrial cytology in mares

Experimental studies of endometrial smears from 26 normal mares showed that after a period of sexual rest exceeding 7 days smears were generally free from inflammatory cells. Smears from 9 mares showing signs of persistent endometritis contained inflammatory cells in 91 per cent while bacterial culture was positive in only 45 per cent. Smears were also taken from 242 Thoroughbred mares as a routine procedure in 1977. It was concluded that endometrial cytology provides a better guide to uterine inflammation than bacteriology. The routine clinical use of endometrial cytology is recommended to aid the interpretation of both positive and negative bacterial cultures from the cervix.     

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.     

Uterine morphology and function in postpartum mares

Ultrasonically detectable characteristics of the uterus and embryo and palpable uterine tone were assessed in 10 postpartum mares. A bright fern-like pattern of ultrasonic uterine echogenicity, outlining the endometrial folds, was observed for an average of 2.1 ±0.2 days following parturition (range, 1 to 3 days). Unexpectedly, the uterus was quiescent throughout the postpartum interval, based on daily one-minute contractility scans. Contractility was maximal on Days 12 to 15 of pregnancy in both postpartum (n=7) and nonparturient (n=7) mares. The mean diameter of ultrasonically detectable intrauterine fluid collections increased (P<0.05) abruptly between days 1 and 2 postpartum and gradually decreased (P<0.05) between days 4 and 7; no collections were detected after day 16. There was no effect of day on echogenicity of the intrauterine fluid collections; on all days, fluid was relatively black or nonechogenic, suggesting that puerperal endometritis was not a problem in this group. Because the increase in intraluminal fluid occurred after parturition and in temporal association with a decrease in diameter and tone of the uterus, the fluid collections apparently represented a physiologic influx from the involuting uterus rather than residual placental fluid. Involution of the horns was completed by day 27 (formerly nongravid horn) and day 31 postpartum (formerly gravid horn), based on failure to detect further significant decreases in diameter. However, the formerly gravid horn was larger (P<0.05) in diameter than the formerly nongravid horn on each of Days 1 to 35 postpartum (end of experiment), indicating residual effects on uterine size. When averaged over both horns, uterine diameters were larger on Days 0 to 24 (Day 0=day of ovulation) of pregnancy in postpartum mares than in nonparturient mares; by Day 25, diameters were similar between statuses. By approximately Day 6 of pregnancy, uterine contractility and ultrasonic endometrial exhotexture were similar between postpartum mares and nonparturient mares. Uterine tone was greater (P<0.05) in postpartum mares than in nonparturient mares on all days between Day 0 and 25. An unexpected, transient increase in uterine tone was detected on Day 5 of pregnancy in both postpartum mares and nonparturient mares. No differences were found between reproductive statuses in patterns of embryo mobility, the day of fixation of the embryonic vesicle (postpartum, Day 15.3 ±0.4; nonparturient, Day 15.0 ±0.3), and diameter of the embryonic vesicle on the day of fixation (postpartum, 22.1 ±1.4 mm; nonparturient, 19.4 ±l.6mm). However, mean uterine tone and mean horn diameters on the side of fixation were greater (cranial and middle cornual segments; P<0.05) or tended to be greater (caudal segment; P<0.1 ) on the day of fixation in postpartum mares than in nonparturient mares. In all postpartum mares, fixation occurred in the formerly nongravid horn. Enhanced uterine tone in postpartum mares may account for the occurrence of fixation on the same day for the two reproductive statuses, despite the larger uterus in postpartum mares.     

Equine post-breeding endometritis: A review

The deposition of semen, bacteria and debris in the uterus of the mare after breeding normally induces a self-limiting endometritis. The resultant fluid and inflammatory products are cleared by 48 hours post cover. Mares that are susceptible to persistent post-breeding endometritis (PPBEM) have impaired uterine defence and clearance mechanisms, making them unable to resolve this inflammation within the normal time. This persists beyond 48 hours post-breeding and causes persistent fluid accumulation within the uterus. Mares with PPBEM have an increased rate of embryonic loss and a lower overall pregnancy rate than those without the condition. To enhance conception rates, mares at high risk need optimal breeding management as well as early diagnosis, followed by the most appropriate treatment. This article reviews the pathogenesis, diagnosis and treatment of PPBEM and the management of affected mares.     

Progesterone and equine chorionic gonadotropin concentrations around the time of pregnancy loss in mares impregnated by donkeys or stallions

The objective of this study was to compare the incidence of pregnancy loss of mares carrying a mule embryo with that of mares carrying a horse embryo. The possible causes of such mortality were evaluated through serial ultrasonographic evaluations and hormonal monitoring, paying special attention to the role of premature regression of the endometrial cups and its relation to inadequate luteal function.

Twenty-eight mares impregnated by stallions and 19 mares impregnated with donkey semen were evaluated ultrasonographically every week from day 20 to day 150 of pregnancy. The viability of the product was assessed each time, and the diameter of the embryonic vesicle was measured from day 25 to day 60. Blood samples for progesterone and equine chorionic gonadotropin (eCG) determination were taken every week.

Both progesterone and eCG concentrations during normal pregnancies wegre significantly lower in the mares inseminated with donkey semen than in the mares impregnated by stallions (P<.05). In 7 of the mares carrying mule conceptuses to term, the concentrations of eCG remained basal throughout the study. In the other animals from this group, the levels of this hormone did increase but returned to baseline much earlier (on day 77 of pregnancy) than in the mares served by stallions (on day 126 day of pregnancy). There was no significant difference between the growth rate of embryonic vesicles of mares carrying mule embryos and that of mares carrying horse embryos (P>.05).

The incidence of pregnancy loss was significantly higher (P<.05) in mares carrying a mule embryo (36.8 %) than in mares carrying a horse embryo (21.4%); it occurred on average on day 93 of pregnancy in mares carrying mule embryos and on day 43 on mares carrying horse embryos. There was only 1 case in which pregnancy loss was associated with concentrations of both eCG and progesterone that were much lower than the average for the normal pregnancies of the same group, and this was in a mare carrying a horse embryo. The most frequent cause of pregnancy loss was premature luteal regression due to primary luteolysis, as evaluated via peripheral progesterone concentrations. This occurred in 2 mares carrying horse embryos and in 4 mares carrying mule embryos. Three mares carrying mule embryos and 1 carrying a horse embryo had abortions that were not preceded or accompanied by any alteration in progesterone or eCG levels and were thus classified as fetal deaths of non-endocrine origin.

It is concluded that the incidence of pregnancy loss is higher in mares carrying a mule embryo than in mares carrying a horse embryo. However, this is not due to the low progesterone concentrations associated with the premature regression of the endometrial cups that occurs in mares with interspecific pregnancy.     

Laparoscopic Application of PGE2 to the Uterine Tube Surface Enhances Fertility in Selected Subfertile Mares

Topical application of prostaglandin E₂ (PGE₂) gel to the surface of the uterine tubes via a laparoscopic procedure improved embryo recovery rates or pregnancy rates in 28 subfertile mares suspected of uterine tubal pathology. Gelatinous masses may occlude the lumen of the uterine tube and prevent sperm from reaching the site of fertilization or prevent embryos from reaching the uterus. PGE₂ is secreted by the early equine embryo, promoting passage of the embryo into the uterus; topical administration of PGE₂ onto the surface of the uterine tube has been shown to stimulate early transport of the embryo into the uterus. Embryos were produced or a pregnancy was obtained from 24 of the 28 barren mares treated with direct laparoscopic application of 0.2 mg of PGE₂ to their uterine tubes. Mares had been barren for an average duration of 1.9 ± 0.6 years and an average of 6.9 ± 3.8 estrous cycles prior to treatment, without donating an embryo or becoming pregnant. Seventeen of 20 mares bred as embryo donors produced one or more embryos with an average of 2.1 ± 1.9 embryos collected per mare (0.45 embryos per cycle) after PGE₂ treatment. Seven of 8 mares bred to carry their own pregnancy became pregnant within the first two cycles following PGE₂ treatment. These 8 mares were bred an average of 5.6 ± 1.8 cycles without a pregnancy prior to treatment. The laparoscopic PGE₂ procedure was performed during various stages of the estrous cycle; the stage varied among treated mares.     

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.     

Escherichia coli Lipopolysaccharide-Induced Immunomodulation Along With Oxytocin Administration After Mating as a Treatment Protocol for Persistent Endometritis in Mares

Endometritis is accepted as a major hindrance to achieve optimal reproductive efficiency in mares. The objective of the present study was to evaluate the therapeutic efficacy of combined therapy of immunomodulator and ecbolic as an alternative stand-alone therapy for mares with persistent endometritis. On the basis of history, culture, endometrial cytology, and per rectal and/or ultrasonographic genital examinations, 76 subfertile mares were selected and assigned to three age groups and four treatment (G-1, 2, 3) and control (G-4) groups. At estrus, all the mares were bred once naturally. Thereafter, the mares of G-1 (n = 28) were aseptically treated at 6-hours after natural service with intrauterine infusion (in 50 mL normal saline solution) of 100 μg of Escherichia coli lipopolysaccharide (LPS). Additionally, these mares received two injections of 20 IU of oxytocin (IV) at 12 and 24-hours after infusion. Mares in group G-2 (n = 11) were treated with LPS as mares of G-1, whereas mares in G-3 (n = 12) received oxytocin injections only. The mares of G-4 (n = 23) did not receive any treatment. Pregnancy rates at day 21 and foaling rates were higher (P < .001) in group G-1 than in G-4. In G-1, higher percentage of mares at ages 6–10 years conceived and foaled than mares aged ≥16 years. On re-swabbing of mares that remained nonpregnant, the majority of G-1 and G-2 mares demonstrated sterile cultures and negative cytology, whereas uterine inflammation persisted in mares of G-3 and G-4. In conclusion, the combined therapy was effective for the elimination of persistent endometritis and improved reproductive performance of subfertile mares.     

Recovery and evaluation of embryos from normal and infertile mares

To evaluate embryo transfer as a possible method to circumvent infertility in mares, embryos from 14 normal and 14 infertile mares were collected three times and examined. Fewer flushes (p less than 0.05) from normal than infertile mares (1/42 vs 9/42) contained only abnormal embryos whereas more flushes (p less than 0.05) from normal than infertile mares contained one or more normal embryos (28/42 vs 8/42). More flushes (p less than 0.05) from normal than infertile mares contained embryos (29/42 vs 17/42). The embryo diameters (mm) at either day-7 or day-8 post ovulation were greater (p less than 0.01) for normal than infertile mares (day 7: 07 +/- 0.08 vs 0.3 +/- 0.07; day 8: 1.1 +/- 0.18 vs 0.7 +/- 0.23). Six of the 10 (60%) flushes that contained only abnormal embryos were recovered from ares with positive uterine cultures or moderate to severe endometritis. The embryos recovered from normal mares were greater in quantity and better in quality.     

Early foetal death in the mare. Histological, bacteriological and cytological findings in the endometrium.

Fifteen mares which had each lost their pregnancies 2 to 8 times during the preceding 6 years were used in a study comprising 1 or 2 breeding seasons for each mare. During the research period all 15 mares conceived and 10 of the mares had normal pregnancies in the 1st experimental year. Five mares resorbed/aborted once or twice during the 1st year. This was followed by a pregnancy that terminated in a live foal. Histopathological examinations of uterine biopsies showed a wide range of histopathological conditions, from absence of changes excessive for the mare's age and parity to the most severe changes of the endometrium. Most likely, endometritis, acute as well as chronic degenerative endometritis, is the most common cause of early foetal death in the mare. Good breeding management is essential and was often enough to make the mares produce a foal. Additional treatment was needed in some of the mares and was based on the course of the conceptus loss and the condition of the endometrium, as determined by clinical, histological, bacteriological and cytological examinations.     

Effect of pre- and postmating nutritional manipulation on plasma progesterone,blastocyst development,and the oviductal environment during early pregnancy in gilts

Two experiments were conducted to determine mechanisms mediating effects of nutritional manipulation before and after mating on embryonic survival in pigs. Experiment 1 studied the mechanisms by which continued high feeding levels after mating result in differences in plasma progesterone during early pregnancy. Gilts fed 2.0 times maintenance energy requirements either remained on this high level or feed was reduced to 1.5 times maintenance immediately after mating. Ovarian, oviductal, and jugular vein progesterone concentrations were determined 72 h after onset of estrus, and samples taken every 4 h were used to determine LH and progesterone during the periestrous period. Treatment did not affect peripheral progesterone concentrations, the timing or rate of rise of progesterone, or progesterone in ovarian, oviductal, or jugular veins at the time of surgery. Time after the LH peak was highly correlated (P = 0.0001) with jugular progesterone concentrations, but not with those in oviductal and ovarian veins, suggesting that responses in the reproductive tract mediated by peripheral progesterone concentrations will be temporally different to effects within tissues supplied by the ovarian and oviductal vasculature. Experiment 2 studied mechanisms mediating nutritional manipulation in the preovulatory period on postovulatory reproductive function, using feed restriction during the first (RH) or second (HR) week of the estrous cycle. Surgeries were performed 12 to 20 h after ovulation, and fertilized oocytes were cultured for 144 h in vitro. Ovulation rate was not affected by previous nutritional regimen. Fertilization rate was higher (P = 0.056) in RH vs HR gilts, but development of cultured oocytes was not affected by treatment. There were no treatment differences in peripheral or oviductal plasma progesterone, estradiol, or insulin-like growth factor-I (IGF-I) at surgery, or in porcine oviductal secretory protein abundance and IGF-I concentrations in oviduct flushings, but treatment affected total protein concentration (P = 0.002). These results indicate that either previous nutritional treatment does not affect the early developmental competence of fertilized oocytes in vitro or differences in developmental competence of oocytes are not expressed up to the early blastocyst stage. However, the lack of an effect of previous nutrition on steroids in the local oviductal circulation may also be related to the lack of effects on oviductal function and embryonic development.     

Reduced quality of bovine embryos cultured in media conditioned by exposure to an inflamed endometrium

OBJECTIVE: A suboptimal uterine environment contributes to the bovine repeat breeder syndrome. Subclinical endometritis is a component, so the mechanism by which inflammation affects embryo survival was investigated by assessing the effect of non-cellular products of an inflamed endometrium on embryo development to blastocyst. DESIGN: Endometrial fluid from a lactating dairy cow was collected by lavage using embryo culture medium. Aseptic inflammation was then induced by infusion of glycogen and lavage was repeated 6 h later. The recovered fluid was used to culture Day 5 in-vitro-derived embryos for 2 days. Embryo development and quality were compared for two treatment groups (culture media conditioned by inflamed or non-inflamed endometrium) and two controls (control or control + serum). RESULTS: Development to blastocyst was higher for conditioned media or media + serum (inflamed 42.2%; non-inflamed 49.3%; control + serum 50.9%; control 16.9%; P < 0.001). Blastocyst cell numbers were lower for the conditioned-inflamed group (inflamed, 83.1; non-inflamed, 99.8; control + serum, 100.6; control, 110.1; P < 0.001). Trophectoderm cell number, but not inner cell mass number, was reduced in the conditioned-inflamed group and the inner cell mass:trophectoderm ratio was increased (P < 0.001). CONCLUSION: Altering the embryo culture environment with the products of endometrial inflammation had a subtle effect on embryo quality. An increased inner cell mass:trophoblast ratio is likely to negatively affect embryonic survival. Altered embryo quality is a mechanism for early embryonic failure in cows with subclinical endometritis. Culture of embryos with normal endometrial fluid may improve in vitro embryo production.     

Assessment of the breeding prognosis of mares using paired endometrial biopsy techniques

Paired endometrial biopsy samples were taken from 530 subfertile mares, before and after treatment (where indicated) and a period of sexual rest. Prognoses were made after each biopsy (Categories 1A-4A before treatment and Categories 1B-4B after treatment), using histopathological criteria similar to those described by Kenney and Doig (1986). Eighty-seven per cent of the mares were assigned to first biopsy prognosis Category 3A. The second biopsy prognosis produced a more even population distribution (10, 47, 40 and 3 per cent respectively for Category 1B, 2B, 3B and 4B mares). First biopsy Category 3A and 4A mares produced mean foaling rates of 51 and 40 per cent respectively. None of the 14 second biopsy Category 4B mares delivered a live foal (12 were barren, one aborted and one suffered early foetal death). This study supports the hypothesis that a breeding prognosis assigned on the basis of two endometrial biopsy samples, taken before and after specific uterine treatment, where indicated, which takes into consideration the mare's ability to respond to treatment, is likely to be more accurate than one assigned after a single biopsy sample.     

Evaluation of clinical and luteolytic effects of a novel prostaglandin analogue in normal and problem mares

Alfaprostal (K 11941), a novel prostaglandin F2 alpha analogue was clinically tested in 24 dioestrous mares, 40 anoestrous mares and 31 postpartum mares, all being given 2 or 3 mg intramuscularly. Blood samples were taken for the determination of plasma progesterone levels and the objective confirmation of luteolytic drug effects. Two hundred and thirty-six mares at the same location were used for comparisons of the rates of pregnancy, early embryonic loss and foaling. Alfaprostol was found to act as a potent luteolytic agent with good oestrus induction, follicular development and normal conception (72.6%) and foaling rate (65.3%). No side effects were observed. Treatments commenced early in the season (October), when only 35% of the anoestrous mares showed elevated progesterone levels, but in 80% of anoestrous mares with baseline progesterone levels, alfaprostol initiated heat and ovulation followed by normal fertility.