A method for the accurate measurement of opsonic activity in uterine secretions of the mare

This study was undertaken to develop a method for accurately measuring opsonic activity in the uterine secretions of the mare. Ten mares were used in the study. They ranged in ages from 6 to 19 years and were of various genital health status. Undiluted uterine secretions were collected by inserting a tampon into the uterus during estrus; serum samples were collected simultaneously Opsonic activity in the secretions and serum was analyzed in a chemiluminescence assay, in which zymosan particles were opsonized. Opsonic activity was determined as peak chemiluminescence, time to peak chemiluminescence, and total chemiluminescence (area under curve). The peak chemiluminescence was 16 to 17 times higher when uterine secretions were used for opsonization rather than when buffer was used. Compared to the opsonic activity in serum, the peak chemiluminescence was 21% (P相似文献   

Hormonal and immunological changes in blood and mammary secretion in the sow at parturition

The purpose of the present study was to record possible variations of estradiol-17 beta (E2) and cortisol concentrations, and parameters related to granulocyte phagocytosis in mammary secretions from healthy sows at parturition. The study was comprised 8 primiparous sows (Landrace x Yorkshire). Blood and mammary secretion samples were collected twice daily from 3 d before (only blood) until 3 d after farrowing. Estradiol-17 beta and cortisol concentrations were determined in plasma and in cell-depleted skimmed mammary secretions. Phagocytic capacity of polymorphonuclear cells (PMN) was assessed in whole blood and in cell suspensions derived from mammary secretions. Opsonic activity was assessed in serum and in cell-depleted skimmed mammary secretions. The 2 assays were based on chemiluminescence. Estradiol-17 beta concentration in plasma decreased (P < 0.001) directly after parturition. In skimmed secretions, the highest E2 concentration was recorded in the first sample after parturition and decreased (P < 0.01) thereafter. The highest cortisol concentration in plasma was recorded in the evening before parturition (P < 0.01). In skimmed secretions, there was no significant variation in cortisol concentration. The concentrations of both steroid hormones were lower in mammary secretions than in plasma. The phagocytic capacity of PMN in blood and mammary secretion, expressed as peak chemiluminescence per PMN, showed no significant change. This was also true for the opsonic activity in serum. In skimmed secretions the opsonic activity increased (P < 0.01) after parturition. These data emphasize the differences between plasma and mammary secretion concentrations of steroid hormones as well as between systemic and mammary gland immune competence. Regarding the phagocytosis process in mammary secretions, the part directly related to the PMN function seemed not to be altered at parturition compared with later on in lactation, whereas the part related to opsonic activity seemed to be impaired at parturition. The latter may play a role in the development of coliform mastitis at this time.     

Immunoglobulins in uterine secretions of mares with differing resistance to endometritis

The immunoglobulins IgA, IgG, IgG(T) and IgM were measured in uterine secretions from mares with normal uterine defense capability against bacterial contamination, and in mares with lowered resistance. Samples were collected for analysis at two stages of estrus and two stages of diestrus. All mares were then challenged with a pathogenic culture of Streptococci inoculated into the uterus. The immunoglobulins were quantitated on a similar schedule post-inoculation. Generally higher amounts of IgA, IgG and IgG(T) were found in the uterine secretions of mares which had an imparied resistance to endometritis than in mares with an efficient defense mechanisms. IgM was not detected in enough samples to suggest any differences.     

Lysozyme, alkaline phosphatase and neutrophils in uterine secretions of mares with differing resistance to endometritis

A study was conducted to 1) determine differences in the inflammatory response following bacterial challenge between normal mares and mares with chronic endometritis and 2) to determine if enzyme activity in uterine fluid can be used to evaluate degree of inflammation in the equine uterus. Six normal mares (Group 1) and four mares with chronic endometritis (Group 2) received an intrauterine infusion of beta-hemolytic streptococci on the second day of estrus. Neutrophil concentration as well as lysozyme and alkaline phosphatase activity were determined in uterine secretions obtained by placing tampons in the uterus of mares. All mares had a similar inflammatory response following bacterial challenge of the uterus, as indicated by a neutrophil response of the same magnitude. Neutrophil numbers, lysozyme and alkaline phosphatase concentrations were all increased 12 h postinoculation and declined rapidly to normal preinoculation values by 48 h after inoculation. In spite of the similarity of the clinical signs, neutrophil concentrations and enzyme activity, mares in group 1 demonstrated a markedly higher ability to eliminate the infection than mares in group 2. It is concluded that factors other than neutrophil numbers, lysozyme and alkaline phosphatase activity account for the inability of the mare to eliminate uterine infections.     

Hastened transport of equine embryos through the oviduct of the mare

The purposes of this experiment were 1) to test the hypothesis that placing rabbit embryos into the mare's uterus would hasten oviduct transport and 2) to determine if placing fluid into the uterus of bred mares on Day 4 and/or Day 5 would subsequently disrupt the mare's pregnancy. The hypothesis that placing rabbit embryos into the mare's uterus would hasten oviduct transport was not supported, since the uterine recovery rate of equine embryos on Day 5 was not significantly higher (P>0.05) for mares receiving rabbit embryos on Day 4 than for mares receiving no uterine infusion on Day 4 (1 10 vs 0 10 , respectively). However, placing fluid into the mare's uterus on Day 4 was apparently responsible for hastened oviduct transport, since mares with media infused into the uterus on Day 4 had a significantly higher (P<0.05) recovery rate of equine embryos on Day 5 than did mares receiving either rabbit embryos or no uterine infusion on Day 4 post ovulation (5 10 vs 1 10 or 0 10 , respectively). The Day-14 pregnancy rate was significantly higher (P<0.05) for mares receiving no uterine infusion on Day 4 or Day 5 than for mares receiving uterine infusion on Day 5 or uterine infusion on both Days 4 and 5 (9 10 vs 4 10 , 2 10 and 0 10 , respectively).     

Effects of long‐distance running on serum opsonic activity measured by chemiluminescence

Exhaustive exercise such as long‐distance running has been shown to increase susceptibility to infection. In order to investigate whether serum opsonic activity plays a role in such conditions, we utilized luminol‐dependent and lucigenin‐dependent chemiluminescence (LmCL and LgCL). We took serum samples from 24 male marathon runners before and after running 30 km. Neutrophils were isolated from the peripheral blood of healthy volunteers. Serum opsonic activity was examined by measuring neutrophil ROS stimulated with zymosan particles opsonized by the serum samples. Immunoglobulin and complement levels in the serum were also measured. After a 30 km run, the maximum light emission was increased and the time to reach the maximum light emission was shortened significantly (p < 0.05) in LmCL. However, there were no significant changes in the immunoglobulin and complement levels. The increase of ROS production may suggest that serum opsonic activity is accelerated after running 30 km. Thus, serum opsonic activity might not play a significant role in the susceptibility to infection after long‐distance running. Copyright © 2003 John Wiley & Sons, Ltd.     

Differences in uterine position of reproductively normal mares and those with delayed uterine clearance detected by scintigraphy

The position of the uterus within the abdomen may affect a mare's ability to rapidly clear the uterine lumen of contamination. In this study, the position of the uterus was determined from left and right lateral flank scintigrams taken 1 and 2 h after intrauterine infusion of radiocolloid. Scintigraphy was performed during estrus in 44 mares, 24 were reproductively normal and 20 exhibited a delay in uterine clearance. Reproductively normal mares were nulliparous (n = 14) or pluriparous (n = 10), 3 to 21 yr of age, had no history of persistent uterine infections and cleared > 50% of a radiocolloid within 2 h of infusion into the uterus. Mares that exhibited a delay in uterine clearance were pluriparous (n = 18) or nulliparous (n = 2), 12 to 24 yr of age, had a history of endometritis and cleared < 30% of a radiocolloid within 2 h. The angle between the caudal-ventral aspect of the uterine image and cervix relative to horizontal as visualized on the scintigram was measured with a protractor. Results were analyzed by the General Linear Model System. The uterine-cervical angle relative to horizontal was more ventral in mares with delay in uterine clearance and was more horizontal in reproductively normal mares (mean +/- SEM-111.6 +/- 3.6 for delay in uterine clearance mares; 147.6 +/- 3.9 for reproductively normal mares; P < 0.0001). The mean angle for reproductively normal, pluriparous mares was steeper than that for nulliparous mares (141.1 +/- 2.9, 152.3 +/- 2.44 respectively; P = 0.004). There were no differences in angles between left and right lateral views within individuals. We conclude that a uterus that tilts ventrally in relation to the pelvic brim may contribute to the inability of delay in uterine clearance mares to rapidly clear their uterine lumen of contamination. Parity may contribute to the more ventral orientation of the uterus.     

Immunoglobulin levels,protein concentrations and alkaline phosphatase activity in uterine flushings from mares with endometritis

Uterine samples and flushings were obtained from 87 mares to compare endometrial bacteriology and biopsy with immunoglobulin and protein concentration and alkaline phosphatase activity in uterine flushings. Mares were designated as infected if both bacteriology and biopsies were positive. The immunoglobulin levels, protein concentration and alkaline phosphatase activity in uterine flushings from infected and non-infected mares were compared. Twenty (23%) of the mares were classified as infected. A significantly higher proportion of infected mares (cf. non-infected) had elevated IgA and protein concentrations. Levels of IgG, IgG_T or alkaline phosphatase were not significantly elevated in infected mares. These results suggest that IgA and protein levels are elevated in the uterus in the presence of active infection.     

The effect of postbreeding uterine lavage on pregnancy rate in mares

One stallion and 54 mares were used in an experiment to evaluate the effect of postbreeding uterine lavage on pregnancy rate in mares. All mares were inseminated with 250 x 10(6) progressively motile sperm every other day during estrus until detection of ovulation. Mares (n = 18) were randomly assigned to one of three treatment groups: 1) no postbreeding uterine lavage (control); 2) uterine lavage at 0.5 h postbreeding; or 3) uterine lavage at 2 h postbreeding. A dilute solution of povidone-iodine (PIS; 0.05%) previously determined to render spermatozoa immotile in vitro was used to lavage the mare uteri. One liter PIS, prewarmed to 40 degrees C, was used for each lavage. Pregnancy status of mares was determined at 21 d and 36 d post ovulation, using transrectal ultrasonography. The pregnancy rate of Group 1 (66.7%) was higher than that of Group 2 (22.2%; P<0.05) or Group 3 (33.3%); P<0.10). The pregnancy rates of Groups 2 and 3 were similar (P>0.70). Evaluation of endometrial biopsies obtained from a separate set of mares (n = 3) on Day 6 post ovulation, both before and after uterine lavage, revealed no difference in the accumulation of inflammatory cells, suggesting adverse effects of lavage on fertility may have been due to excessive removal of spermatozoa from the uterus during the lavage process or damage to oviductal spermatozoa.     

Changes in intrauterine pressure after oxytocin administration in reproductively normal mares and in those with a delay in uterine clearance

Intrauterine pressure was measured in 4 reproductively normal mares and 4 mares with delay in uterine clearance after administration of oxytocin to determine if intrauterine pressure varied between dosage and group. Changes in intrauterine pressure were measured during estrus, when a follicle was > or =35 mm, using a Millar "Mikro-tip" catheter that had 3 discrete pressure sensors/channels. Mares received 4 different treatments of 10, 5, 2.5 or 0 IU (vehicle) of oxytocin. The protocol for each treatment consisted of a 10-min baseline recording, administration of treatment and measurement of changes in intrauterine pressure for 65 min. After administration of the first two treatments, mares were rested for 2 h and the protocol repeated for the remaining 2 treatments. Changes in intrauterine pressure were measured on a physiograph and stored in a computer. The results were analyzed by 4x4 Latin Square Design analysis of variance (ANOVA) using the GLM procedure of the Statistical Analysis System. The ANOVA detected a main effect of treatment (P<0.01) and mare (nested within group; P<0.01) but no effect of channels, group or treatment-by-group interaction. There was a dose-dependent increase in uterine activity in both normal mares and those with delayed uterine clearance. A dose of 10 IU of oxytocin induced a larger number of uterine contractions (5.67+/-0.06) for a longer time (24.09+/-1.18 min) than the 5 IU (4.16+/-0.06 contractions and 16.31+/-1.18; P<0.01 min) or 2.5 IU dose (4.08+/-0.06 contractions and 17.61+/-1.18 min). The first intrauterine wave occurred most often near the tip of the horn in 10 of 12 recordings in normal mares and in 8 of 12 recordings in mares with delayed uterine clearance. It was then propagated from the middle of the horn to the uterine body just cranial to the cervix. There was no pattern of propagation for subsequent intrauterine pressure waves. We conclude that the difference in spontaneous clearance of the uterus between the 2 groups is not reflected in their response to exogenous oxytocin as determined by changes in intrauterine pressure.     

Purification and characterization of a humoral opsonin from the solitary urochordate Styela clava.

1. We have previously identified opsonic activity in the plasma of the solitary urochordate, Styela clava. 2. Here, we report the purification and further characterization of the opsonic molecule. 3. Two purification methods were employed. 4. Gel filtration yielded one strongly opsonic fraction that contained a single, electrophoretically-resolved protein. 5. Opsonic activity was dose-dependent and sensitive to tryptic digestion and heat denaturation. 6. SDS-PAGE and calibrated gel filtration indicated the opsonic protein was a 17.5 kDa monomer while isoelectrofocusing indicated a single pI of 7.0. 7. In an alternative procedure, a similar opsonic activity and protein were isolated by affinity purification using whole yeast cells.     

Effects of estrous cycle and season on ultrasonic uterine anatomy in mares

The morphological changes in ultrasound images of the uterus at various times of the year were characterized in nonbred mares, using a linear-array scanner. The uterus was recorded as having an ultrasonic morphology characteristic of diestrus (uterine score 1, endometrial folds not visible), estrus (score 3, distinct endometrial folds), or an intermediate stage (score 2). In Experiment I, uterine scores for the first ovulatory period of the year were compared to scores for the second period in 23 pony mares. More mares (P<0.05) showed endometrial folding prior to the second ovulation of the year (14 23 ) than prior to the first (5 23 ). Mean uterine scores were higher (P<0.05) on Day -10 (ovulation = Day 0) and tended to be higher (P<0.1) on Days -14, -13, and -11 of the first ovulatory period than on the corresponding days of the second period. Uterine scores for the first ovulatory period were lower (P<0.05) on Days -5, -4, -3, -2, -1, and 0 and tended to be lower (P<0.01) on Day -6. In addition, the pattern of change in uterine scores paralleled the pattern of change in the intensity of estrous behavior. In Experiment II, in 20 horse mares, the curve for uterine scores during interovulatory intervals in May-June, but not in September-October, was bimodal due to a small rise (P<0.05) and subsequent return to baseline between Days +3 and +6. The mean uterine scores for both May-June and September-October began to increase on Day -7 or -8, reached maximum on Day -3, declined between Days -2 and 0, continued to decline after Day 0, and reached a value characteristic of diestrus by Day +2. Results indicated that the ultrasonic characteristics of the uterus may provide an instant indicator of estrogen exposure and may have practical value in judging the optimal time to breed.     

Interaction between equine semen and the endometrium: the inflammatory response to semen.

Insemination of mares with bacteria-free equine spermatozoa results in an influx of polymorphonuclear neutrophils (PMNs) into the uterine lumen. In vitro studies have demonstrated that equine spermatozoa activate complement, resulting in cleavage of factors C5a and C3b. Since uterine secretion is rich in complement, it is likely that an interaction between spermatozoa and uterine secretion results in C5a-mediated chemotaxis and migration of PMNs into the uterine lumen. Once in the uterine lumen, the PMNs phagocytize bacteria and spermatozoa, which is an important part of sperm elimination from the reproductive tract. It is not clear how the spermatozoa are opsonized, or if phagocytosis of equine spermatozoa is a selective or non-selective process. Breeding-induced endometritis appears to be both up and down regulated by seminal components. A modulatory role on the inflammation has been suggested for equine seminal plasma. Seminal plasma suppressed complement activation, PMN-chemotaxis and phagocytosis in vitro. Preliminary in vivo experiments also support a suppressive role of seminal plasma in breeding-induced endometritis. The duration but not the magnitude of the PMN-influx into the uterine lumen was shortened when seminal plasma was included in an insemination dose. The presence of PMNs in the uterus affects the motion characteristics of spermatozoa in vitro. Both progressive motility and mean path velocity were impaired when spermatozoa were incubated in uterine secretion from mares with ongoing breeding-induced endometritis. The binding of spermatozoa to PMNs was prominent in all samples collected from mares with an ongoing endometritis. The motility remained impaired, but the binding of the spermatozoa to PMNs was reduced when the spermatozoa were incubated in uterine secretion in the presence of seminal plasma. Preliminary characterization of the immune-suppressive component in seminal plasma suggests that it is one or more molecule(s) with a molecular weight between 50 and 100 kDa, partially inactivated by charcoal stripping and partially heat-inactivated at 95 degrees C for 45 min.     

Opsonic activity of antisera to ribosomal vaccine fractions with live and formalinized Pseudomonas aeruginosa

The opsonic capacity of antisera to Pseudomonas aeruginosa ribosomal vaccine fractions was determined by a chemiluminescent technique. Antiserum to a vaccine fraction ("peak A") containing lipopolysaccharide (antiserum A), and antiserum to a vaccine fraction ("peak B"), which did not contain detectable amounts of lipopolysaccharide (antiserum B), were used to opsonify live or formalin-treated bacteria. Polymorphonuclear leukocytes were then stimulated by the opsonified bacteria in the presence of the chemiluminigenic probe, luminol, resulting in the observed chemiluminescence. The data obtained indicated that the antisera had comparable opsonic activity with live (untreated) bacteria. However, antiserum B had far less opsonic activity than did antiserum A when formalinized bacteria were used. Owing to the effects of formaldehyde on protein, these results were interpreted as evidence to suggest that the opsonic activities of the two antisera are dependent on different antigens on the bacterial cell surface. Antiserum A activity is probably dependent on lipopolysaccharide to a great extent, whereas antiserum B activity is most likely dependent primarily on a protein(s).     

Characterization of intrauterine mobility of the early equine conceptus

Intrauterine mobility patterns of the embryonic vesicle were characterized on Days 9 to 17 after ovulation in pony mares using real-time ultrasonography (n=5 or 7 mares per day). The location of the vesicle was determined by dividing the uterus into right horn, left horn, and body. Each uterine horn was further divided into three approximately equal portions (cranial third, middle third, caudal third), yielding seven segments (body plus three portions of each horn). Location of the vesicle within the uterus was recorded every five minutes for two consecutive hours (25 location determinations per trial). The number of times the vesicle was found in the uterine body versus one of the uterine horns was greater for the body on Day 9 (15.2 vs 9.8; not significant) and Day 10 (17.3 vs 7.7 P<0.05) and greater (P<0.05) for the horns on Days 12 (7.3 vs 17.7) through 17 (0.0 vs 25.0). Averaged over all days, when the vesicle was in one of the uterine horns it was present 56% of the time in the caudal third, 30% of the time in the middle third, and 14% of the time in the cranial third. Mobility was determined by the number of times the vesicle changed locations during successive examinations. On Day 9, the mean number of location changes per trial was minimal (horn to horn, 0.2; body to horn or vice versa, 1.8; between two segments, 4.2). The extent of mobility increased on Day 10 and reached an apparent plateau from Day 11 to Day 14. The mean number of location changes per trial during the plateau was as follows: horn to horn, 1.6; body to horn or vice versa, 5.6; between two segments, 10.7. Fixation (cessation of mobility) occurred in one of the horns in 5 7 mares on Day 15 and in 7 7 mares by Day 16. Mobility was present on the earliest day the embryonic vesicle was detected (Day 9), but Days 11 to 14 were characterized as the days of maximum mobility.     

Uterine contractile activity in mares during the estrous cycle and early pregnancy

Transrectal ultrasonography was used to quantitate uterine contractile activity during the estrous cycle and early pregnancy in pony mares (nonbred, n = 9; pregnant, n = 16). Continuous 1-min scans of longitudinal sections of the uterine body were videotaped, and uterine activity scores (1=minimal activity, 5=maximal activity) were assigned to each tape segment. There was a tendency (P<0.06) for a main effect of reproductive status (nonbred versus pregnant), a main effect of day (P<0.0001), and a reproductive status by day interaction (P<0.006). Uterine activity scores were higher (P<0.05) in pregnant mares on Days 1, 11, 12, and 17 (Day 0=day of ovulation) than in nonbred mares. Maximal activity in pregnant mares occurred on Days 11 to 14 during the reported period of maximal embryo mobility. Activity scores decreased (P<0.05) between the day prior to and the day of fixation (mean = Day 15) of the embryonic vesicle. Activity scores were maintained at an intermediate level for several days following fixation before declining to minimal levels by 7 d postfixation. A postovulatory decrease (P<0.04) in activity scores was observed in nonbred mares, but not in pregnant mares, between Days 0 and 1 followed by a progressive increase (P<0.03) between Days 2 and 4. Maximal activity in nonbred mares occurred during the late luteal phase (Days 13 to 14), corresponding temporally to the reported onset of luteolysis.     

Effects of cervical dilation and intrauterine infusions on the timing of oviductal transport of equine embryos

Equine embryos spend 5 to 6 d in the oviduct before entering the uterus as expanded blastocysts, and cannot be consistently collected nonsurgically until Day 7. Technologies such as cryopreservation and embryo splitting, which are most successful with embryos at the morula or early blastocyst stage, have not been used in mares because equine morulae and early blastocysts are located in the oviduct and cannot be recovered nonsurgically. These experiments test the hypothesis that transport of equine embryos through the oviduct can be hastened by cervical dilation or by acute, sterile endometritis induced by intrauterine oyster glycogen treatment. Cervical dilation with or without intrauterine infusion of 0.5 ml PBS on Day 4 did not appear to hasten the transport of embryos into the uterus since Day 5 uterine embryo recovery rates were not higher (P > 0.1) for mares with cervical dilation or cervical dilation plus PBS infusion vs mares receiving no treatments (0 of 5 and 0 of 5 vs 0 of 10, respectively). Intrauterine infusions of 40 ml of 1% oyster glycogen or 40 ml of PBS on Day 3 did not appear to hasten the transport of embryos into the uterus since Day 5 uterine embryo recovery rates were not higher (P > 0.1) for oyster glycogen- or PBS-treated vs untreated mares (2 of 12 and 3 of 11 vs 0 of 10, respectively). Cervical and uterine treatments on Day 3 or Day 4 and uterine lavages on Day 5 decreased (P < 0.05) Days 11 to Day 15 pregnancy rates compared with that of untreated mares. Day 11 to Day 15 pregnancy rates were 1 of 5 for mares with Day 4 cervical dilation and Day 5 uterine lavage, 1 of 5 for mares with Day 4 PBS infusion and Day 5 uterine lavage, 2 of 12 for mares with Day 3 oyster glycogen infusion and Day 5 uterine lavage, and 3 of 11 for mares with Day 3 PBS infusion and Day 5 uterine lavage vs 7 of 10 for mares that received no treatment or lavage. Cervical and uterine manipulations on Day 3 or 4 and uterine lavage on Day 5 appeared to decrease pregnancy rates by Days 11 to 15. The results of these experiments do not support the hypothesis that cervical dilation or uterine infusion hasten oviductal transport, since neither cervical manipulation nor transcervical infusion of oyster glycogen or PBS into the uterus significantly hastened the rate of embryo transport into the uterus.     

Effects of day of estrous cycle, time of day, luteolysis, and embryo on uterine contractility in mares

One-minute continuous ultrasonic scans of longitudinal sections of the uterine body were videotaped, and contractility scores (1 to 5, minimal to maximal contractility) were assigned without knowledge of mare identity, day of the estrous cycle or pregnancy status. Contractility was assessed, and plasma progesterone concentrations were determined for each of 3 daily examinations (at 0800, 1600 and 2400 hours) from Day 9 to Day 19 (Day 0 = day of ovulation). For both the nonbred (n=11) and pregnant (n=11) mares, there was no effect of hour of scan on the extent of uterine contractility. When data for the nonbred mares were normalized to the onset of luteolysis (defined for each mare as the first >/=25% decrease in plasma progesterone concentrations between successive samples), there was an abrupt increase (P<0.05) in contractility 24 hours prior to the onset of luteolysis. Contractility was also assessed daily in 20 nonbred and 27 pregnant mares from Day 0 to Day 17. For the nonbred mares, a biphasic profile in contractility occurred during the estrous cycle as indicated by the following significant changes: a decrease between Days 0 and 2, an increase between Days 2 and 4, a plateau between Days 4 and 7, a decrease between Days 7 and 11, an increase between Days 11 and 13, and a decrease between Days 14 and 16. For pregnant mares, contractility increased (P<0.05) prior to the late-diestrous increase for nonbred mares. In addition, a significant reduction in contractility was detected on Day 5 in these mares compared with that in the nonbred mares. Contractility in the uterine body in 7 mares was assessed every 5 minutes after departure of the embryonic vesicle from the uterine body. Levels of contractility in the uterine body were lower (P<0.05) 55 minutes after the vesicle had exited the body than 相似文献   

Uterine infusion of conceptus fragments changes the protein profile from cyclic mares

This experiment aimed to compare at day seven after ovulation, the protein profile of uterine fluid in cyclic mares with mares infused two days before with Day 13 conceptus fragments. Experimental animals were ten healthy cyclic mares, examined daily to detect ovulation (Day 0) as soon as estrus was confirmed. On day seven, after ovulation, uterine fluid was collected, constituting the Cyclic group (n = 10). The same mares were examined in the second cycle until ovulation was detected. On day five, after ovulation, fragments from a previously collected concepti were infused into each mare''s uterus. Two days after infusion, uterine fluid was collected, constituting the Fragment group (n = 10). Two-dimensional electrophoresis technique processed uterine fluid samples. A total of 373 spots were detected. MALDI-TOF/TOF and NanoUHPLC-QTOF mass spectrometry identified twenty spots with differences in abundance between the Cyclic and Fragment group. Thirteen proteins were identified, with different abundance between groups. Identified proteins may be related to embryo-maternal communication, which involves adhesion, nutrition, endothelial cell proliferation, transport, and immunological tolerance. In conclusion, conceptus fragments signalized changes in the protein profile of uterine fluid seven days after ovulation in comparison to the observed at Day 7 in the same cyclic mares.     

Work in progress: A simple technique that may improve the rate of embryo recovery on uterine flushing in mares

Embryo recovery rates from uterine flushings of normal mares on Day 7 or later after ovulation currently range from 55% to 80%. In contrast, pregnancy rates at 14 d in experimental mares are often higher. There appears to be a discrepancy between pregnancy rates and recovery rates of embryos on uterine flushing, indicating that some embryos are not recovered from the uterus on flushing. Per rectum ultrasound examination of the uterus of mares during flushing suggested that in some mares, the infused fluid may accumulate in the uterine body and not extend to contact the entire uterus, even after massage of the filled uterus per rectum. To increase embryo recovery rates, the flusing technique was altered to allow 3 min contact time of the flush fluid with the uterus during each of three flushes. It was thought that during this time, if the embryo was not directly contacted by the infused fluid, mobility of the embryo might cause it to move into the fluid, and thus be collected. This technique was used in 20 flushes on 14 mares, from 7 to 11 d after ovulation. Embryos were recovered on 18 of the 20 flushes. A total of 21 embryos was recovered, for an embryo recovery rate of 105%. The recovery rate from mares with single ovulations was 13/15 (87%); the recovery rate from mares with multiple ovulations was 8/5 (160%). These rates appear to be higher than those obtained previously in our laboratory and those reported by other workers in the field. These results indicate that further investigation into the efficacy of this procedure is warranted.