Qualitative analysis and functional classification of the uterine proteome of mares in oestrus and dioestrus.

Quantitative analysis of the uterine flush fluid proteome of mares in oestrus and dioestrus has been previously reported. The objectives of this study were to: a) evaluate qualitative differences in the uterine flush fluid proteome between mares in oestrus and mares in dioestrus and b) perform a functional classification of proteins either unique to each stage or common between the two stages. Uterine flush fluid samples were collected from 8 light breed mares in either oestrus (n = 5) or dioestrus (n = 3). Proteomic analysis of the samples was conducted using liquid chromatography-tandem mass spectrometry. Proteins exclusively detected in oestrus or dioestrus and those common to both stages were identified using the Scaffold software (version 4.4.8, Proteome Software Inc., Portland, OR). The identified proteins were classified into gene ontology (GO) categories (cellular component [CC], molecular function [MF] and biological process [BP]) using the PANTHER (www.pantherdb.org) classification system version 14.0. Of 172 proteins identified, 51 and 28 were exclusively detected in mares in oestrus and dioestrus, respectively, and 93 proteins were common to both stages. The most represented terms in various GO categories were similar among the three subsets of proteins. The most represented CC terms were extracellular region and cell, the most represented MF terms were catalytic activity and binding, and the most represented BP terms were metabolic process and cellular process. In conclusion, proteomic analysis of the uterine flush fluid enabled the identification of subsets of proteins unique to oestrus or dioestrus, or common to both stages. The results of this study can serve as a baseline for future research focused on finding stage-specific protein markers or evaluating differences in the uterine flush fluid proteome between normal mares and those with uterine disease.

Analysis of the uterine flush fluid proteome of healthy mares and mares with endometritis or fibrotic endometrial degeneration.

The objective of this study was to evaluate the differences in the uterine flush fluid proteome between healthy mares and mares with endometritis or fibrotic endometrial degeneration (FED). Uterine flush fluid samples were collected from healthy mares (n=8; oestrus n=5 and dioestrus n=3) and mares with endometritis (n=23; oestrus n=14 and dioestrus n=9) or FED (n=7; oestrus n=6 and dioestrus n=1). Proteomic analysis was performed using label-free liquid chromatography-tandem mass spectrometry. Of 216 proteins identified during oestrus, 127 were common to all three groups, one protein was exclusively detected in healthy mares, 47 proteins were exclusively detected in mares with endometritis and four proteins were exclusively detected in mares with FED. Of 188 proteins identified during dioestrus, 113 proteins were common between healthy mares and mares with endometritis, eight proteins were exclusively detected in healthy mares and 67 proteins were exclusively detected in mares with endometritis. Quantitative analysis revealed a subset of proteins differing in abundance between the three groups during oestrus and between healthy mares and mares with endometritis during dioestrus. These results provide a springboard for evaluation of specific proteins as biomarkers of uterine health and disease and for investigation of their roles in the establishment and maintenance of pregnancy.

The impact of ecbolic therapy in the early postpartum period on uterine involution and reproductive health in dairy cows.

Postpartum uterine disease due to poor uterine involution continues to be a significant factor that contributes to poor reproductive efficiency in dairy cattle. Therapy that increases the frequency, duration and strength of uterine contractions in the postpartum period might enhance uterine involution, resulting in improved reproductive performance. The objective of this clinical trial was to study the effect of two uterine ecbolic therapies, oxytocin and prostaglandinF2α on uterine involution, postpartum endometritis, and reproductive performance. A randomized double-blinded clinical trial was conducted in 118 dairy cows from two research herds that had normal parturition and expulsion of the fetal membranes. Within 24 hr after calving, cows were randomly assigned to receive intramuscular injections twice a day of 50 IU of oxytocin, or 25 mg of dinoprost (PGF2α) or saline (control) for 7 days. Cows were monitored from day 3 to day 63 ± 3 days postpartum by transrectal palpation of the uterus, vaginoscopy, Metricheck® examination and by endometrial cytology. Blood samples were collected for measurement of progesterone in weeks 3 (21 ± 3 days), 5 (35 ± 3 days), 7 (49 ± 3 days), and 9 (63 ± 3 days) postpartum. Herd breeding records were obtained to determine reproductive performance in all cows. Neither oxytocin nor prostaglandin F2α therapy during the first week postpartum had any significant effect on the rate of uterine involution, prevalence of endometritis or reproductive performance, compared to untreated controls. Ecbolic drugs, as used here, are not recommended for use in clinical practice to improve involution or reproductive tract health in normal cows.

Treatment of Hydropsical Conditions Using Transcervical Gradual Fetal Fluid Drainage in Mares With or Without Concurrent Abdominal Wall Disease.

Hydropsical conditions are exceedingly rare in the horse. However, when they occur, they are true emergencies due to the severe enlargement of the pregnant uterus, which can result in clinical signs, such as an enlarged round abdomen, dyspnea, reluctance to walk, and colic, and may lead to the development of abdominal wall disease. The pathogenesis of hydropsical conditions is not fully elucidated, but they have been associated with placentitis and fetal abnormalities. This report describes six cases of hydropsical conditions in mares with or without concurrent abdominal wall disease. Five out of six cases were hydrallantois, and of these five, two mares had abdominal wall disease; the remaining one out of six cases was hydramnios. All mares were treated by termination of the pregnancy through gradual fluid drainage transcervically over a number of hours, and their fetuses were delivered vaginally. All fetuses were euthanized immediately after vaginal delivery. Of the six mares, two had signs of placentitis, two were confirmed seropositive for leptospirosis, and two were euthanized (one because of a vaginal tear that communicated through the peritoneum and one mare that developed abdominal wall rupture and laminitis). The remaining 4 mares were available for follow-up; three mares were not rebred, and one mare became an embryo donor, with a successful embryo recovery. We reported the prevalence of leptospira involvement in two out of six cases of hydrallantois and also described the clinical outcome of the mares after treatment with slow fetal fluid drainage.

Dose-dependent inhibition of uterine contractility by nitric oxide: A potential mechanism underlying persistent breeding-induced endometritis in the mare.

Nitric oxide (NO) may have a role in persistent breeding-induced endometritis in mares through an inhibitory effect on uterine contractility. The objectives of this study were to test the effect of NO on spontaneous uterine contractility in-vitro and to evaluate whether this effect varied between the longitudinal and circular muscle layers of the uterus. Reproductive tracts were collected from eight euthanized non-pregnant mares (age 4-19 years; body weight 405-530 kg). Transrectal examination of the reproductive tract was performed before euthanasia to evaluate stage of the estrous cycle and presence of any apparent abnormality. After euthanasia, one uterine tissue sample was collected for histological evaluation and four full-thickness uterine tissue strips (10-12 mm × 2 mm), two parallel to each muscle layer, were excised for in-vitro contractility evaluation. Strips were suspended in tissue chambers containing Krebs-Henseleit solution, with continuous aeration (95% O2-5% CO2; pH 7.4) at 37 °C. After equilibration, spontaneous contractility was recorded (pre-treatment) and strips excised in each direction were randomly allocated to each of two groups: 1) SNAP (S-nitroso-N-acetylpenicillamine, an NO donor); or 2) NAP (N-acetyl-d-penicillamine, vehicle and time-matched control). These were treated at 15 min intervals with increasing concentrations (10-7 M to 10-3 M) of SNAP and NAP, respectively. Contractility data was recorded throughout the experiment. An interaction effect of group-by-concentration was observed (P < 0.0001). The mean contractility after treatment with 10-4 M and 10-3 M SNAP were significantly lower than the pre-treatment contractility and the mean contractility after treatment with lower SNAP concentrations. In contrast, contractility did not change significantly in the NAP treated controls. The effect of treatment on uterine contractility was not influenced by age or weight of the mare, stage of estrous cycle, uterine histology grade, or muscle layer. Secondary findings included significant main effects of stage of estrous cycle (increased contractility in estrus compared to diestrus), uterine histology grade (decreased contractility in grade IIB compared to grade I) and age (decreased contractility in mares aged > 8 years compared to mares aged ≤ 8 years). In conclusion, results of this study indicate that NO has a dose-dependent inhibitory effect on spontaneous uterine contractility irrespective of the muscle layer in the mare.

Persistence of oxytocin receptors in the bovine uterus during the first 7 d after calving: an immunohistochemical study.

Persistence of the oxytocin receptor (OTR) in the bovine uterus during the first 7 d after calving was investigated by means of immunohistochemical staining. Immunoreactive OTRs were present in different locations in the uterus on almost all days except day 2, when staining was seen only in the endothelium of blood vessels in the endometrium and myometrium. This finding supports the hypothesis that oxytocin may be ecbolic in cows through the 1st week post partum, but further studies are required to assess the receptor functionality during this period.

La persistance de récepteurs pour l'ocytocine (OTR) dans l'utérus bovin durant les 7 premiers jours suivant le vêlage a été examiné par coloration immunohistochimique. Des OTR immuno-réactifs étaient présents dans différentes localisations dans l'utérus à presque tous les jours sauf au jour 2, alors que la coloration ne fut notée que dans l'endothélium des vaisseaux sanguins dans l'endomètre et le myomètre. Cette trouvaille soutient l'hypothèse que l'ocytocine pourrait être ocytocique chez les vaches durant la 1ère semaine post-partum mais des études supplémentaires sont requises pour évaluer la fonctionnalité des récepteurs durant cette période.(Traduit par Docteur Serge Messier).

Foaling rates and risk factors for abortion in pregnant mares presented for medical or surgical treatment of colic: 153 cases (1993-2005).

The purpose of this study was to determine foaling rates in mares presented for medical or surgical treatment of colic, and to examine risk factors associated with abortion following colic. A retrospective analysis of 153 medical records found that mares treated surgically for colic (P = 0.0007) were 3.5 times more likely to have a negative pregnancy outcome than were mares treated medically for colic. Anesthetic time (P = 0.01) and intra-operative hypotension (P = 0.03) were significantly associated with negative pregnancy outcome. Mares with an anesthetic time > or = 3 h were 6 times more likely to abort. Signs of endotoxemia (P = 0.30), hypoxia (P = 0.89), flunixin meglumine administration (P = 0.13), mucous membrane color at the time of presentation (P = 0.82) and capillary refill time (P = 0.76) were not associated with pregnancy outcome. There was no difference in the foaling rate for mares that had received progestin supplementation versus those that had not (P = 0.42). In this study, the significant risk factors for abortion were surgically treated colic, long anesthetic time, and intraoperative hypotension.