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Query: UMLS:C0016382 (flushing)
 6,387 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

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.
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PMID:Work in progress: A simple technique that may improve the rate of embryo recovery on uterine flushing in mares. 1672 90

Embryos were collected from superovulated donor cows on Day 7 of the cycle either in vivo by a standard nonsurgical method (A) or in vitro from the excised uterus after slaughter of the donor cow (B). In Method B, the time between slaughter and flushing of the uterus ranged from 0.5 to 4 hours. Flush yield was 5.6 +/- 5.0 and 8.4 +/- 5.3 embryos (P<0.01); the recovery rate was 0.6 +/- 0.4 and 0.8 +/- 0.3 (P<0.05) for Methods A and B, respectively. Method B resulted in more Grade 3 (P<0.001) and 4 embryos, while Method A resulted in more Grade 1 and 2 embryos. The correlation between the percentage of Grade 1 and 2 embryos and the time interval between slaughter of the donor cow and flushing the excised uterus was -0.42 for in vitro flushes. Viability of fresh and frozen-thawed embryos, as determined by in vitro culture of Grade 1 and 2 morulae and early blastocysts, was considerably lower for Method B than Method A. The percentage of embryos developing into expanded blastocysts was 100% (10 10 ) and 40% (4 10 ) for fresh embryos (P<0.01) and 52.7% (29 55 ) and 0% (0 25 ) for the frozen-thawed embryos (P<0.001) for Methods A and B, respectively. This reduction in viability might be the result of a postmortem pH decrease in the uterine fluids within the first 30 minutes from 7.0 to 5.8 and 6.0. Flushing of the uterus directly after slaughter (within 5 to 10 minutes) may prevent the possible detrimental effect of a low pH on the embryos.
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PMID:Number and viability of embryos collected in vivo or from the excised uteri of slaughtered donor cows. 1672 90

In Experiment 1, 12 unmated cyclic ewes received twice-daily intrauterine injections on Days 12 to 14 of one of the following treatments: 1) ovine conceptus secretory proteins (oCSP) containing 25 microg of ovine trophoblast protein-1 (oTP-1) as determined by RIA; 2) 25 or 50 microg recombinant human interferon alpha1 (rhlFN); or 3) 1500 microg of serum proteins (oSP) from a Day-16 pregnant ewe (estrus = Day 0) per uterine horn. Ewes receiving oCSP had longer interestrous intervals (27 +/- 2 days; P<0.05) than ewes receiving oSP (17 +/- 2 days). Ewes receiving either dose of rhlFN had an interestrous interval of 16 +/- 2 days which did not differ (P>0.10) from that of oSP-treated ewes. In Experiment 2, 59 normally cycling ewes, mated on Day 0, received twice-daily intramuscular injections of either 2 mg recombinant bovine interferon alpha1 (rblFN) or placebo on Days 12 to 15 post estrus. On Day 16, pregnancy was confirmed by flushing a morphologically normal conceptus from the uterus. Pregnancy rates for rblFN-treated (80%) and placebo-treated (62%) ewes were not different (P>0.10). Uterine flushings and conceptus-conditioned medium were assayed for oTP-1. Total oTP-1 in conceptus-conditioned culture medium was higher (P<0.02) when conceptuses were from placebo-treated (104 +/- 14 microg/conceptus) than from rblFN-treated (56 +/- 12 microg/conceptus) ewes; while total oTP-1 in uterine flushings was similar (P>0.10) for placebo-treated (132 +/- 15 microg/conceptus) and rblFN-treated (147 +/- 17 microg/conceptus) ewes. The interval from mating to subsequent estrus following conceptus removal was 31 +/- 1 and 28 +/- 1 days for pregnant ewes treated with rblFN and placebo, respectively. Interestrous intervals for nonpregnant ewes were longer (P<0.02) for rblFN-treated (27 +/- 3 days) than for placebo-treated (18 +/- 2 days) ewes.
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PMID:Effect of recombinant alpha interferons on fertility and interestrous interval in sheep. 1672 86

Variability in the superovulation response is an important problem for the embryo transfer industry. The objective of this study was to determine whether FSH treatment at the beginning of the cycle would improve the ovulation rate and embryo yield in dairy cows. Twenty-eight postpartum cyclic dairy cows were allocated at random to 4 treatment groups (A, B, C and D). Group A cows (n=10) received FSH (35 mg) at a decreasing dose, starting on Day 9 (Day 0=day of estrus) for 5 days followed by PGF2alpha (35 mg) on Day 12. Cows assigned to Groups B, C and D (n=6 cows each, respectively) were given 35 mg FSH at a decreasing dose from Days 2 to 6 followed by PGF2alpha on Day 7. Group C and D cows received PRID inserts from Day 3 to Day 7. Cows in Group D additionally received 1000 IU hCG 60 hours after PGF2alpha treatment. Ovaries were scanned daily using a real time ultrasound scanner from the beginning of FSH treatment until embryo recovery, to monitor follicular development, ovulation and the number of unovulated follicles. Embryos were recovered from the uterus by a nonsurgical flushing technique 7 days after breeding. There were no differences (P>0.01) in the number of follicles>10 mm at 48 hours after PGF2alpha treatment among the 4 groups. The mean numbers of follicles were 10.6+/-1.2, 9.3+/-1.3, 12.2+/-1.3 and 15.0+/-2.9 for Groups A, B, C and D, respectively. A significantly (P<0.001) higher number of ovulations was observed and a larger number of embryos was recovered in Group A than in the other groups. The results of this study indicate that superovulation with FSH at the beginning of the cycle causes sufficient follicular development but results in very low ovulation and embryo recovery rates.
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PMID:Follicular growth, ovulation and embryo recovery in dairy cows given FSH at the beginning or middle of the estrous cycle. 1672 12

A total of 141 embryos was recovered by nonsurgical flushing of the uterus of 31 superovulated buffalo. A total of 66 good quality embryos (Grade I and Grade II) was frozen using 1.4 M glycerol. Forty-two of the frozen embryos were thawed randomly over a 1-year period, and a total of 39 embryos (Grades I, II or III post thaw) were transferred into an equal number of estrus synchronized recipients. Of 11 confirmed pregnancies, 9 calves were born live.
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PMID:Successful culmination of pregnancy and live birth following the transfer of frozen-thawed buffalo embryos. 1672 86

Beginning on Day 10 or 11 of the estrous cycle, mature Holstein heifers were given a superovulatory regimen of twice-daily injections of porcine FSH, together with injections of PG with the fifth and sixth FSH injections. Every 12 h from 24 to 60 h after PG administration, the animals received im injections of different doses of the LH releasing hormone antagonist [N-Ac-D-Nal(2)(1), D-pCl-Phe(2), D-Trp(3), D-Arg(6), D-Ala(10)]-LHRH or vehicle. Follicular development was monitored by transrectal ultrasonography every 12 h from 24 to 120 h after PG administration. All animals were given hCG at 72 h after PG injection, and were artificially inseminated. At Day 7 of gestation, the corpora lutea were counted by ultrasonography, and embryos were collected by nonsurgical flushing of the uterus. Treatment with the antagonist resulted in a dose-dependent decrease in the amplitude of the LH surge and in delays in the time of occurrence of the LH surge, ovulation and the shift from estradiol to progesterone production. These results indicate that LHRH antagonists can be used to delay the LH surge and ovulation in superovulated heifers. This finding may be beneficial to studies in the superovulation of cattle.
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PMID:Effects of an LHRH antagonist on the time of occurrence and amplitude of the preovulatory LH surge, progesterone and estradiol secretion, and ovulation in superovulated Holstein heifers. 1672 48

We describe the first complete embryo transfer program, including flushing of embryos from the oviducts via the uterine horns, transfer of embryos into the Fallopian tubes or the uterine horns and recording of the number of piglets born live. The described procedure is minimally invasive and allows the use of pigs simultaneously for embryo collection and production of normal pregnancies. A 30 degrees forward oblique endoscope provided optimal visualization of the reproductive organs and free access to the organs for embryo flushing and transfer. In contrast to surgical and nonsurgical methods, endoscopy allows to pre-examine the genital tract for reproductive abnormalities and successful ovulation. A total of 95 prepuberal gilts or cyclic sows were used in this trial. Embryos or oocytes were collected from hormonally treated pigs via endoscopy(n = 17) on Day 3 and via laparotomy or post mortem after slaughter (control group, n = 38) on Day 3 and 6 after insemination. One (unilateral collection, n = 7) or both oviducts (bilateral collection, n = 10) were flushed endoscopically. We recovered 114 (average 16/pig) and 279 (average 28/pig) oocytes or embryos with fertilization rates of 89% and 72%, respectively. In the control group 834 oocytes or embryos were collected at Day 3 and 6 after insemination (fertilization rate 64%, total 534 embryos, 33 at 2-, 367 at 4-, 2 at 8-cell stage, 24 morulae and 108 blastocysts). Of 836 embryos recovered by endoscopy, surgery or slaughter 528 Day 3 embryos at 2- to 4-cell stage were transferred into (one) oviducts (n = 27 pigs, about 20/pig) resulting in 9 pregnant pigs diagnosed at Day 28 by sonography. Of the 9, 8 carried a total of 49 piglets to term. A total of 195 Day 6 embryos were transferred into uterine horns (n = 12 pigs, about 16/pig), resulting in 5 pregnant pigs carrying a total of 38 offspring to term. The use of endoscopy in assisted reproduction of pigs has the advantages of allowing easy access to the ovary, oviduct and uterus, clear view of the organ manipulation without exposure and exteriorization of viscera during surgery.
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PMID:Endoscopic embryo collection and embryo transfer into the oviduct and the uterus of pigs. 1672 55

The objective of our study was to determine the effect of conditioning media with homologous porcine uterine cells on the developmental rate of porcine embryos. Cell monolayers were prepared by selective dissection and digestion of sections from the uterus of prepuberal gilts that were primed with PMSG and hCG. Conditioned media were used with 2 type of embryos: 4-cell stage (Experiment 1) or blastocyst stage (Experiment 2). In Experiment 1, embryos were collected surgically by flushing the oviducts, 36 to 48 h following the first of 2 inseminations. Embryos were cultured in Whitten's medium containing 1.5% BSA as a protein source until they attained the 4-cell stage. Embryos at the 4-cell stage were cultured randomly in either Whitten's medium with 1.5% BSA or Whitten's medium with 1.5% BSA that was previously conditioned for 24 h with an endometrial epithelial cell monolayer. Embryos were cultured in 50-microl drops covered with oil in a 38.5 degrees C, 5% CO(2) in air incubator. There was no advantage to using the conditioned media with the 4-cell stage embryos. The embryos were less developed than those cultured in nonconditioned Whitten's medium (P <0.001). In Experiment 2, embryos were cultured at the blastocyst stage. They were recovered the same way as in Experiment 1 and then cultured in Whitten's medium containing 1.5% BSA until they reached the blastocyst stage. At the blastocyst stage (Day 6), embryos were randomly assigned to 1 of the 6 following treatments: Whitten's with 1.5% BSA or Whitten's plus 1.5% BSA that was previously conditioned with endometrial epithelial cell monolayer, TCM-199 containing 0.4% BSA or TCM-199 plus 0.4% BSA that was previously conditioned with endometrial epithelial cell monolayer, finally, TCM-199 containing 10% serum or TCM-199 plus 10% serum that was previously conditioned with endometrial epithelial cell monolayer. Results show that initiation of hatching was significantly enhanced by conditioning the Whitten's media.
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PMID:Effects of conditioned media on porcine embryos at different stages of development. 1672 81

Our objective was to determine whether the concentration of cooled sperm inseminated influenced sperm transport and intensity of the uterine inflammatory reaction 2, 4 and 24h after insemination. Experimental subjects were 189 estrous mares with a dominant follicle > or =35 mm in diameter and no bacterial growth or neutrophils detected in uterine smears. Each mare was randomly assigned to receive one of the following intrauterine treatments (volume, 20 mL): insemination with 5x10(6) mL(-1) or 25x10(6) mL(-1) or 50x10(6) mL(-1) sperm diluted in 3 mL seminal plasma (SP) and 17 mL skim milk; seminal plasma or skim milk extender. Mares in a control group received no intrauterine treatment. Mares were slaughtered 2, 4 or 24h after insemination or infusion. Oviducts were separated from the uterus, and uterus and oviducts were then flushed with phosphate-buffered saline (PBS). After flushing, an endometrial sample was collected for further histopathological examination. The grade of uterine fibrosis and the amount of neutrophils in the stratum compactum were evaluated. A sample of each tubal flushing was examined for sperm count, and a sample of each uterine flushing was examined for PMN count. It was concluded that compounds in the insemination dose provoked a uterine inflammatory response, which was more rapid and intense as sperm concentration increased. In contrast, sperm transport through 4h after insemination was not influenced by sperm concentration.
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PMID:Effect of sperm numbers and concentration on sperm transport and uterine inflammatory response in the mare. 1703 41

The aim of this study was to compare (a) two different umbilical cord blood (UCB) collection methods while the placenta is still in the uterus (in utero), and (b) to evaluate the efficacy of four cryopreservation protocols based on UCB haematopoiestic stem cell (HSC) recovery. We analysed UCB samples collected with our original collection system designed for active Syringe/Flush/Syringe method or by standard in utero method. For comparing different cryopreservation procedures, dimethyl sulphoxide (DMSO) at final concentration of 5 and 10% was used and combined with our own controlled-rate or uncontrolled-rate cryopreservation. A total of 99 samples were collected. A significantly higher UCB volume, total nucleated cell and mononuclear cell were seen following the first collection strategy (n= 49; mean +/- SD, 103 +/- 35.4 mL; 12.34 +/- 5.27 x 10(8); 595 +/- 3.47 x 10(6)) vs. the second strategy (n= 50; 86 +/- 29.3 mL; 9.87 +/- 4.47; 424 +/- 2.82 x 10(6)) respectively (P < 0.01). The discard rate was 14% for the first and 36% for the second collection strategy (P < 0.01). It was shown that the most efficient procedure was the controlled-rate protocol combined with lower (5%) DMSO concentration. Using active Syringe/Flush/Syringe method, we collected UCB with greater volumes and with lower discard rate compared to the standard by gravity technique. The data presented also showed much better recovery of UCB cells when controlled-rate freezing procedure and 5% DMSO were combined.
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PMID:Collection strategies and cryopreservation of umbilical cord blood. 1743 Apr 66


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