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Biomedical subjects

G S Lewis

Publications and source records attributed to G S Lewis.

At least 19 recordsLinked to original sources
PubMed2006

Effect of early and late exposure to estrual ewes on ram sexual performance classifications.

This study was conducted to determine whether exposure of ram lambs to estrual ewes during their first autumn and again as adults just before serving capacity tests (SCT) affected the outcome of the sexual performance tests. Treatments were either early exposure of Polypay ram lambs (i.e., 7-8-mo-old rams with ewes for 17 d [n=30] or no early exposure [n=30]), and late exposure (i.e., 16-19-mo-old rams with estrual ewes for 3 d) or no exposure to estrual ewes in a 2x2 factorial arrangement. Three serving capacity tests were conducted immediately after the early exposure period for individual ram lambs that were exposed to ewes early. Three sham sexual performance tests (i.e., four ram lambs placed in test pens for 30-min without ewes) were conducted with ram lambs that were not exposed to ewes early. All rams were evaluated during nine 30-min serving capacity tests over a 2-mo period at 16-19 mo of age to determine sexual performance. Prior to serving capacity tests, one half of the rams from each early exposure treatment were exposed to estrual-induced ewes for 3 d. Specific sexual behaviors (e.g., sniffs, flehmens, foreleg kicks, vocalizations, mount attempts, mounts, and ejaculations) were recorded during serving capacity tests. Number of sniffs, flehmens, foreleg kicks, vocalizations, and mount attempts were summed without estimating the value of importance and analyzed as courtship behaviors. Sexual performance data were analyzed with Mixed model procedures for repeated measures. During serving capacity tests, the early exposed rams exhibited more courtships (40.3+/-8.0 versus 23.4+/-4.6; P<0.05; LSM+/-estimated SE), mounts (11.3+/-1.0 versus 7.7+/-0.9; P<0.01), and ejaculations (3.3+/-0.2 or 2.4+/-0.2; P<0.01) than rams not exposed to ewes as ram lambs, respectively. We conclude that early exposure of 7-8-mo-old ram lambs to estrual ewes improves sexual performance in serving capacity tests at 16-19 mo of age in most rams whereas, late exposure to estrual ewes for 3 d prior to serving capacity tests did not improve sexual performance scores.

Animals↗
PubMed2006

Relationship between sexual behavior classifications of rams and lambs sired in a competitive breeding environment.

The objectives for this study were to 1) determine the relationship between sexual performance class and lambs sired in a competitive mating environment, and 2) determine whether the male-oriented ram test is valid. Fifteen 2- to 3-yr-old whitefaced rams classified as female-oriented, with high or low sexual performance, or classified as male-oriented were used in a multiple-sire breeding arrangement. Five groups of approximately 200 ewes each were exposed for 21 d to 3 rams per group consisting of 1 ram from each class. Rams were blocked for sexual class, and those with close genetic relationship were assigned to different pens. Genomic DNA was prepared from blood collected from 15 rams, 934 ewes, and 1,757 lambs. Up to 4 microsatellite markers were used to determine a lamb's sire. Of 884 ewes with identifiable lambs (known sires), 178 ewes had single lambs, 408 had multiples sired by 1 ram, and 298 had multiples sired by more than 1 ram. The sexual partner preference test used to identify male-oriented rams did not absolutely reflect their sexual performance during competitive breeding. In contrast to only mounting and servicing males in preference tests before breeding, male-oriented rams sired 480 lambs from 330 ewes. Serving capacity tests predicted sexual performance of high and low sexual performance rams. High performance rams impregnated more ewes (499 vs. 258; P < 0.05) and sired more lambs (756 vs. 357; P < 0.05) than did low performance rams, respectively. Low performance and male-oriented rams did not differ for ewes impregnated or lambs sired. We conclude that 1) sexual partner preference tests used to classify male-oriented rams were not absolute in reflecting their breeding performance in a competitive breeding environment; 2) serving capacity tests predicted that high performance rams would breed more ewes than low performance rams and sire more lambs than either low performance or male-oriented rams; and 3) under the conditions of this study, low performance and male-oriented rams did not have an adverse impact on the overall breeding outcome. Combined, low performance and male-oriented rams sired 81 more lambs than did high performance rams, but this required twice as many rams to obtain approximately equal breeding results. Therefore, we suggest that serving capacity tests should be used to select high performance rams, reduce number of rams with marginal sexual performance, and make decisions on ram numbers needed.

Animals↗
PubMed2005

The incidence of genotypes at codon 171 of the prion protein gene (PRNP) in five breeds of sheep and production traits of ewes associated with those genotypes.

Scrapie is one of several transmissible spongiform encephalopathies of livestock. Disease susceptibility is linked to polymorphisms in the normal prion protein gene that encodes the mammalian prion precursor. Codon 171 of this gene is a major determinant of scrapie susceptibility. Selection for arginine (R) at codon 171 is encouraged by the USDA to decrease the incidence of scrapie. Objectives of this study were to determine the frequency of R allele variants at codon 171 in a sample of sheep from five breeds (Columbia, Hampshire, Rambouillet, Suffolk, and Targhee) and western white-faced commercial ewes and to determine whether the R allele is associated with ewe and lamb production traits. Genotyping was performed on 532 ewes and 901 lambs from the University of Wyoming flock, in addition to 820 rams from 52 sheep producers from Wyoming and surrounding areas, using a DNA mismatch assay that discriminated the R allele from others at codon 171. Genotyping was performed by DNA sequencing on 127 rams representing all breeds, except Hampshire from the USDA Sheep Experiment Station at Dubois, ID. The 171R allele was found in all five breeds and in the commercial western white-faced ewes. Genotype frequencies varied (P < 0.001) by breed in ewe and ram populations. Influence of R-allele frequency on ewe lambing records and individual lamb records was analyzed for Columbia (62, 161, 121), Hampshire (89, 193, 162), Rambouillet (87, 179, 133), Suffolk (67, 178, 161), and commercial sheep (227, 463, 324) for numbers of ewes, total number of ewe production records, and individual lamb records, respectively. Suffolk ewes without the R allele (non-R/non-R) gave birth to more (P or= 0.08) by ewe genotype. Lamb birth and weaning weights were not influenced (P >or= 0.12) by lamb genotype in any of the breeds or in the commercial flock. In this population, ultimate lamb production was only influenced by genotype at codon 171 in the Suffolk flock.

Animals↗
PubMed2004

Steroidal regulation of uterine immune defenses.

Progesterone suppresses uterine immune defenses and predisposes postpartum animals to nonspecific uterine infections. Progesterone can also suppress uterine eicosanoid synthesis. This effect of progesterone seems to be an important factor in the onset of uterine infections because eicosanoids can enhance uterine immune defenses. In fact, exogenous prostaglandin F(2alpha) (PGF(2alpha)), an eicosanoid that stimulates uterine PGF(2alpha) production, enhances uterine immune defenses and promotes the ability of ewes and sows to resolve uterine infections, even when progesterone is maintained at luteal phase concentrations. Prostaglandin F(2alpha) is also a proinflammatory molecule that stimulates the production of proinflammatory cytokines and may enhance uterine production of leukotriene B(4) (LTB(4)), which stimulates various neutrophil functions. Neutrophils seem to mount the initial response to bacteria that enter the uterus, and proinflammatory cytokines and LTB(4) enhance phagocytic activity of neutrophils. Even though there are clear associations among PGF(2alpha), LTB(4), proinflammatory cytokines, phagocytosis, and the ability of the uterus to resist or resolve infections, the mechanisms of action of exogenous PGF(2alpha) in mitigating the immunosuppressive effects of progesterone have not yet been defined. However, defining the PGF(2alpha) mechanisms should yield important new information that can be used to develop novel prevention and treatment strategies that do not rely on antibiotic and antimicrobial compounds for managing uterine infections.

Animals↗
PubMed2003

Role of ovarian progesterone and potential role of prostaglandin F2alpha and prostaglandin E2 in modulating the uterine response to infectious bacteria in postpartum ewes.

In sheep and cattle, the postpartum uterus is resistant to bacterial challenge until after corpora lutea develop. A 2 x 2 factorial arrangement of treatments was used to determine whether prostaglandins may mediate the effects of progesterone in transforming the postpartum uterus from resistant to susceptible. On d 14 postpartum, ewes (n = 6/group) were ovariectomized or sham ovariectomized, and the vena cava was catheterized for daily collection of uteroovarian-enriched blood. From d 15 to 20, ewes received twice daily intramuscular injections of progesterone in sesame oil or plain sesame oil. On d 20, each uterus received 75 x 10(7) cfu of Arcanobacterium pyogenes and 35 x 10(7) cfu of Escherichia coli. Uteri were collected on d 25 and examined for signs of infection. For each blood sample, unstimulated and mitogen-stimulated lymphocyte proliferation was measured as [3H]thymidine incorporation, smears were prepared for differential white blood cell (WBC) counts, and progesterone, prostaglandin F2alpha, (PGF2alpha), and prostaglandin E2 (PGE2) were quantified. All 12 progesterone-treated, but only two of the 12 oil-treated, ewes developed uterine infections (P < 0.001). Progesterone treatment increased (P < 0.001; 3.1 vs 1.5 ng/mL) and ovariectomy decreased (P < 0.001; 3.7 vs 0.9 ng/mL) vena caval progesterone. Progesterone treatment reduced (P < 0.01) PGF2alpha, (303.9 vs 801.3 pg/mL), and PGF2alpha was greater (P < 0.05) before than after inoculation (626.4 vs 478.8 pg/mL). The PGE2 concentration was greater in progesterone-treated, ovary-intact ewes than in ewes in the other groups (ovariectomy x progesterone treatment; P < 0.01). Ovariectomy increased (P < 0.005; 4.4 vs 2.9 pmol) and progesterone treatment decreased (P < 0.05; 3.2 vs 4.1 pmol) concanavalin A-stimulated lymphocyte proliferation. Ovariectomy increased lipopolysaccharides-stimulated proliferation (P < 0.05; 2.4 vs 1.9 pmol). For neutrophils per 100 WBC, the ovariectomy x progesterone and progesterone x period interactions were significant (P < 0.01). The ovariectomy x progesterone interaction was significant (P < 0.01) for lymphocytes per 100 WBC. Ovariectomy decreased monocytes (P < 0.001; 10 vs 13) and increased eosinophils (P < 0.001; 10 vs 5) per 100 WBC. Progesterone makes the postpartum uterus in ewes susceptible to infection, but ovariectomy allows ewes to remain resistant; uterine prostaglandins may mediate this change. This model creates opportunities to determine the mechanisms responsible for the shift from resistance to susceptible.

Actinomycetales Infections↗
PubMed2003

Progesterone increases susceptibility of gilts to uterine infections after intrauterine inoculation with infectious bacteria.

In cattle and sheep, a progestogenated uterus is susceptible to infections, but this is not well documented for pigs. Therefore, the effects of day of the estrous cycle and progesterone on the susceptibility to uterine infections were evaluated. Gilts (n = 5 per group) were assigned to treatments in 2 x 2 factorial arrays. In Exp. 1, day of cycle and bacterial challenge were main effects. On d 0 or 8, uteri were inoculated with either 70 x 10(7) cfu of Escherichia coli and 150 x 10(7) cfu of Arcanobacterium pyogenes in PBS or with PBS. In Exp. 2, ovariectomy (OVEX) and progesterone treatment were main effects. On d 0, gilts were ovariectomized or a sham procedure was performed. After surgery, gilts received i.m. injections of progesterone (10 mg/5 mL) or 5 mL of safflower oil diluent twice daily. On d 8, gilts were inoculated with the same doses of bacteria as in Exp. 1. In Exp. 1 and 2, vena caval blood was collected for 4 d, after which uteri were collected. Sediment and ability to culture E. coli and A. pyogenes from uterine flushings were used to diagnose infections. Differential white blood cell counts and lymphocyte response to concanavalin A (Con A) and lipopolysaccharides (LPS) were used to measure lymphocyte proliferation. Progesterone, estradiol-17beta, prostaglandin F2alpha, (PGF2alpha), and prostaglandin E2 (PGE2) were measured in vena caval blood. In Exp. 1, d-8 gilts receiving bacteria developed infections, but d-0 gilts receiving bacteria did not. Daily percentages of neutrophils and lymphocytes changed (P < 0.05) with cycle day and bacterial challenge. Basal- and Con A-stimulated lymphocyte proliferation were greater (P < 0.05) for d-0 than for d-8 gilts. Concentrations of PGF2, (P < 0.01) and PGE2 (P < 0.05) increased after bacterial challenge, regardless of stage of the estrous cycle at the time of inoculation. In Exp. 2, OVEX decreased (P < 0.001) and progesterone treatment increased (P < 0.001) progesterone concentrations, and OVEX decreased (P < 0.01) estradiol-17beta. Gilts with ovarian and/or exogenous progesterone developed infections. Daily percentages of neutrophils and lymphocytes changed in response to OVEX, and neutrophils changed (P < 0.05) in response to endogenous and exogenous progesterone. Lymphocyte proliferation in response to Con A and LPS increased (P < 0.05) with OVEX and decreased (P < 0.05) with progesterone treatment. We conclude that endogenous and exogenous progesterone reduce the ability of the uterus in gilts to resist infections.

Actinomycetaceae↗
PubMed2002

Relationship between postpartum changes in 13, 14-dihydro-15-keto-PGF2alpha concentrations in Holstein cows and their susceptibility to endometritis.

Uterine infections (i.e., endometritis) can have a major economic impact on dairy production. Identifying cows that are susceptible to endometritis and improving the diagnosis of endometritis could lead to a reduction in the impact of such infections. Thus, we used Holstein cows to determine whether postpartum changes in 13, 14-dihydro-15-keto-PGF2alpha (PGFM), a metabolite of PGF2alpha, could be used to identify cows that are susceptible to endometritis and to improve the diagnosis of endometritis. Cows were assigned to three treatments. 1) Control (n = 10) had no clinical or bacteriological signs of endometritis during the study. 2) Treated (n = 11) developed endometritis spontaneously and were treated i.m. with 25 mg of PGF2alpha immediately after clinical diagnosis (d 17.6 +/- 0.8 postpartum; mean +/- SEM). 3) Untreated (n = 10) developed endometritis spontaneously and were not treated after diagnosis (d 20.0 +/- 0.5). Examinations of external and internal genitalia and bacteriological data were used to diagnose endometritis. From d 0 (calving) until approximately d 63 postpartum, jugular blood was collected three times weekly. Progesterone and PGFM were quantified in plasma. For PGFM, the treatment x day interaction was significant (P < 0.01). Overall PGFM profiles for Control and Treated differed (P < 0.05), but the Untreated profile did not differ from either Control or Treated. To better understand the interaction, PGFM data from d 0 to 35 postpartum were partitioned into consecutive 7-d periods, and d-36 and greater data were partitioned into one period. Effects of treatment, day, and the treatment x day interaction were then evaluated within period. Except for the d-15 to -21 period, PGFM was greater (P < 0.03) in Control than in Treated and Untreated. In Treated and Untreated, PGFM increased during the d-15 to -21 period. For progesterone, treatment did not affect the profiles, but day was significant (P < 0.001). Progesterone concentrations were basal from d 0 until approximately d 12, and they generally increased after d 12. Onset of endometritis was associated with increased progesterone concentrations. Treatment did not affect the interval from calving to first detected estrus (29.5 +/- 4.9 d) or from calving to AI (73.3 +/- 8.7 d). We conclude that PGFM measures have the potential to be used to identify cows that are more likely to develop endometritis and that PGFM may aid in the diagnosis of endometritis.

Animals↗
PubMed2001

Scalings and structures in turbulent Couette-Taylor flow.

The scaling of velocity structure functions in Couette-Taylor flow [Lewis and Swinney, Phys. Rev. E 59, 5457 (1999)] is revisited to obtain more accurate values of the scaling exponents for the Reynolds number range investigated, 12,000 to 540,000 (Taylor Reynolds numbers, 34 10(5); this transition corresponds to a visually observed break up of the Taylor vortex roll structure with increasing R.

Journal Article↗
PubMed2001

Measuring nonextensitivity parameters in a turbulent Couette-Taylor flow.

We investigate probability density functions of velocity differences at different distances r measured in a Couette-Taylor flow for a range of Reynolds numbers Re. There is good agreement with the predictions of a theoretical model based on nonextensive statistical mechanics (where the entropies are nonadditive for independent subsystems). We extract the scale-dependent nonextensitivity parameter q(r,Re) from the laboratory data.

Journal Article↗
PubMed2001

Technical note: Artificial vagina vs a vaginal collection vial for collecting semen from rams.

The time required to train rams to an artificial vagina (AV) makes collecting semen from large numbers of rams difficult. To manage this problem, we developed a glass, round-bottomed, 1.9-cm i.d. x 9.8-cm long vaginal collection vial (VCV). Three experiments were conducted to determine whether the VCV affected 1) semen volume per collection, 2) percentage of motile spermatozoa, 3) forward progressive motility score before and after extension and after freezing and thawing, and 4) our ability to collect semen from untrained rams. A soft rubber cap with a hole in the center was used to cover the VCV. A VCV was inserted into the vagina of an estrual ewe, and a monofilament line attached to the VCV was clipped to the wool near the vulva. Rams were joined with unrestrained ewes in a pen until they ejaculated into the VCV. In Exp. 1, five rams trained to an AV were used in a switchback design with four collection periods. During each period (1 d), semen was collected with an AV and a VCV. Immediately after collection, semen volume and sperm motility were quantified. Semen was extended with an aloe vera gel-based diluent at a 1:4 dilution rate, motility was quantified again, and semen was frozen. At 1 h after freezing, semen was thawed and sperm motility was quantified. Ejaculate volume (mean = 0.7 mL) and all measures of motility after collection were similar (P > 0.05) for the two collection methods. In Exp. 2, 10 rams trained to an AV were used in a switchback design with five collection periods (period = 3 d). On d 1 and 3 of each period, an AV and a VCV were used to collect semen. Collection method did not affect (P > 0.05) ejaculate volume (mean = 1.0 mL), percentage of motile cells, or forward progressive motility score. In Exp. 3, 51 untrained rams were used in a switchback design with a single collection period (2 d). Semen was collected with an AV and a VCV. Ability to collect an ejaculate and time required for collection were recorded. The likelihood of collecting semen from untrained rams was greater (P < 0.01) using a VCV (mean = 31.4%) than using an AV (mean = 9.8%). Collection method did not affect (P > 0.05) ejaculate volume (mean = 0.8 mL), percentage of motile cells, or forward progressive motility score. We concluded that a VCV could be used to collect semen from rams that are not trained for semen collection without decreasing ejaculate volume or sperm motility.

Animals↗
PubMed2001

Oxytocin-induced cervical dilation and cervical manipulation in sheep: effects on laparoscopic artificial insemination.

The difficulty of cervical penetration severely limits the use of transcervical AI (TAI) in sheep, and trauma from cervical manipulation (CM) may reduce fertility after TAI. We investigated the effects of cervical dilation using exogenous oxytocin (OT) to facilitate TAI and its effects on reproductive variables after laparoscopic AI (LAI). Estrus was synchronized by inserting pessaries impregnated with 6alpha-methyl-17alpha-hydroxyprogesterone acetate (60 mg) for 12 d. In Exp. 1, we determined whether OT and CM before LAI affected the interval from pessary removal to ovulation and fertilization rate. Crossbred ewes (n = 16) were assigned to 1) saline-CM or 2) OT-CM. In Exp. 2, effects of OT and CM on lambing rates were evaluated with white-faced ewes (n = 220) in a 2 x 2 factorial experiment: 1) saline-sham CM; 2) saline-CM; 3) OT-sham CM; and 4) OT-CM. In both studies, eCG (400 IU i.m.) was injected at pessary removal, and LAI was performed 48 to 52 h later. In Exp. 1, ewes received i.v. either 400 USP units of OT or 20 mL of saline at 30 to 60 min before LAI, and CM was administered as for TAI. Beginning 32 h after pessary removal and continuing at 8-h intervals, ovaries were examined with ultrasonography to estimate time of ovulation. Treatment in Exp. 1 did not affect combined ovum/embryo recovery rate (69%), but OT-CM decreased fertilization rate (47 vs 59%; P < 0.05). The OT tended to reduce the interval to ovulation (OT, 59 h vs saline, 66 h; P < 0.06). The OT x CM interaction in Exp. 1 was not significant. For Exp. 2, approximately 25 min before sham CM or CM, 200 USP units of OT or 10 mL of saline was injected i.v. The LAI was performed immediately after sham CM or CM. At 10 to 12 d after AI in Exp. 2, ewes were mated with Suffolk rams. Blood was collected between 24 and 26 d after AI for pregnancy-specific protein B (PSPB) RIA. The PSPB pregnancy and lambing rates were both 62% in saline-sham controls. The CM did not affect pregnancy (69%) or lambing rate (64%). The OT treatment decreased (P < 0.05) PSPB pregnancy (59%) and lambing rates (56%) in OT-sham ewes and pregnancy and lambing rates in CM ewes (both 43%). Neither CM nor OT before LAI affected lambing rates to next estrus, indicating no long-term damage to the cervix or uterus. In summary, CM did not affect fertility after LAI, but OT decreased lambing rate independent of CM. If OT will not be usable for TAI, it may still be a tool for training TAI personnel.

Animals↗
PubMed2001

Intervals from norgestomet withdrawal and injection of equine chorionic gonadotropin or P.G. 600 to estrus and ovulation in ewes.

Synchronization of estrus and ovulation is essential for AI of ewes during a predetermined time frame, and progestogen-eCG treatments are typically used to prepare the ewes. However, eCG is not readily available in the United States, but P.G. 600 (400 IU of eCG and 200 IU of hCG) is available. Thus, we conducted a study to determine the effects of eCG and P.G. 600 on the timing of estrus and ovulation after progestogen withdrawal. Ewes were assigned to two replicates of an experiment with the following treatments: 1) 3-mg norgestomet implant (i.e., one-half of a Syncro-Mate-B [SMB] implant) for 10 d, plus 2 mL of saline i.m. at SMB removal (n = 11); 2) 3-mg SMB implant for 10 d, plus 400 IU of eCG i.m. at SMB removal (n = 13); and 3) 3-mg SMB implant for 10 d, plus P.G. 600 i.m. at implant removal (n = 9). On d 6 after SMB insertion, PGF2alpha was used to induce luteolysis. Beginning 12 h after implant removal, vasectomized rams were used at 12-h intervals to check for estrus. When a ewe was detected in estrus, each ovary was evaluated ultrasonically. Ovaries were evaluated again 16 h later and then at 8-h intervals until ovulation. Treatment altered the interval from implant removal to estrus (less [P < 0.05] in SMB + eCG than in the other two groups) and to ovulation (greatest [P < 0.05] in SMB). However, the treatment x replicate interaction was significant for the intervals from implant removal to estrus (P < 0.03) and from implant removal to ovulation (P < 0.05). An inconsistent response in the SMB-treated ewes seemed to be primarily responsible for the interaction. The intervals to estrus and to ovulation for the SMB-treated ewes were shorter (P < 0.05) in Replicate 1 than in Replicate 2. Also, both intervals seemed to be less consistent between replicates for the SMB + P.G. 600- than for the SMB + eCG-treated ewes; that is, eCG seemed to increase the predictability of the intervals to estrus and to ovulation. Neither the main effects of treatment and replicate nor their interaction were significant for the interval from estrus to ovulation (38.4 /- 3.3 h), size of the ovulatory follicle (7.7 +/- 0.8 mm), or ovulation rate (1.6 +/- 0.2). We concluded from this experiment that eCG is a better choice than P.G. 600 as the gonadotropin to use at the time of progestogen withdrawal to prepare ewes for AI during a predetermined interval.

Animals↗
PubMed1999

Technical note: development of a transcervical oocyte recovery procedure for sheep.

An oocyte recovery procedure was developed and evaluated to determine whether a transcervical embryo recovery procedure is feasible with our method, which includes estradiol-17beta (E2) and oxytocin (OT) treatments, for dilating the cervix in ewes. On d 6 of an estrous cycle, oocytes were recovered either transcervically or with a laparotomy procedure. In the laparotomy group, ovulation rate was determined during the procedure and was used to calculate the percentage ofoocytes recovered. The laparotomy procedure was a standard uterine flush, and 12 mL of PBS was used to flush each uterine horn. In the transcervical group, the ovaries in each ewe were evaluated ultrasonically to determine ovulation rate. For transcervical recovery, 100 microg of E2 were injected i.v. on d 5 to increase cervical OT receptors, and 100 USP units of OT were injected i.v. 10 to 12 h later to dilate the cervix. Approximately 25 min after OT, ewes were placed in dorsal recumbency in a Commodore cradle, and a modified Foley catheter was passed through the cervix and into the uterus for injection (80 to 210 mL) and aspiration of PBS. The PBS was aspirated with a vacuum pump. The percentage of PBS recovered was greater (P<.01) at laparotomy than with the transcervical procedure (85.8 vs. 36.2%). Despite that difference, oocyte recovery did not differ significantly between the two groups (67% for laparotomy vs. 50% for transcervical; [oocytes recovered/number of corpora lutea] x 100), and there was no evidence that the transcervical procedure damaged the oocytes; the zona pellucida remained intact around all of the oocytes. In conclusion, a procedure that includes E2-OT-induced cervical dilation, passage of a modified Foley catheter into the uterus, and incremental infusion and aspiration of media through the catheter can be used to recover oocytes transcervically from ewes. This procedure may make transcervical embryo recovery feasible for sheep.

Animals↗
PubMed1999

Estradiol-17 beta-oxytocin-induced cervical dilation in sheep: application to transcervical embryo transfer.

Experiments were conducted to determine whether exogenous estradiol-17beta (E2) and oxytocin (OT) can be used to improve transcervical (TC) embryo transfer (ET) procedures for sheep. Our concerns that the E2-OT treatment may alter luteal function prompted Exp. 1, in which 32 ewes were assigned to treatments in a 2x2 factorial array. On d 7 after onset of estrus, ewes received i.v. either 100 microg of E2 or diluent; 12 h later, ewes received i.v. either 400 USP units of OT or saline. To monitor luteal function, progesterone was measured in jugular blood collected from d 7 to 18. The treatments did not affect progesterone concentrations. Two trials were conducted in Exp. 2. In Trial 1, ewes were assigned to one of three treatments: TC transfer with E2-OT treatment to dilate the cervix, laparoscopic ET with E2-OT treatment, or laparoscopic ET with an equivalent diluent that did not dilate the cervix. In Trial 2, ewes were assigned to treatments in a 2x2 factorial array: TC or laparoscopic ET on d 6; E2-OT treatment for cervical dilation or diluents on d 6. Transferred embryos were recovered on d 12 in Trial 1 and d 14 in Trial 2, evaluated morphologically for development, and scored. Treatments did not affect the percentage of transferred embryos recovered. However, mode of transfer decreased (P<.01) the mean embryo development score. The E2-OT treatment increased (P<.01) the development score of embryos transferred transcervically, indicating that cervical dilation may improve the chances of embryos surviving after TC transfer. In conclusion, E2-OT treatment did not affect luteal function, and the E2-OT treatment can be used to enhance the success of TC embryo transfer in sheep.

Animals↗
PubMed1997

Fertility and ovum fertilization rate after laparoscopic or transcervical intrauterine artificial insemination of oxytocin-treated ewes.

Based on our previous work, we found that exogenous oxytocin induces uterine tetany and cervical dilation, and permits transcervical access to the uterus. However, the oxytocin does not reduce sustained sperm transport from the uterus to the oviducts. Thus, we hypothesized that exogenous oxytocin may be a useful adjunct to transcervical intrauterine AI procedures for sheep: two experiments were conducted to test our hypothesis. In Experiment 1, purebred ewes (n = 75/group) were artificially inseminated intrauterine with either laparoscopic or oxytocin-transcervical (i.e., 200 USP units of oxytocin 30 min before AI) procedures. At 54 h after progestogenated pessaries were removed, ewes were inseminated with 200 x 10(6) sperm/0.25 ml of fresh, extended semen, which was collected from a purebred ram of the corresponding breed. Pregnancy rate was greater (P < 0.05) after laparoscopic (37.5%) than after transcervical AI (0%). Because of the disappointing results of Experiment 1, Experiment 2 was conducted to determine whether oxytocin or the AI procedure per se reduced ovum fertilization rate. Treatments were designed in a 2 x 2 factorial arrangement. At 60 h after norgestomet implant removal and 10 min before either laparoscopic or transcervical (cervical in a saline group) AI with 100 x 10(6) sperm/0.25 ml, ewes (n = 10/group) received an intravenous injection of either isotonic saline or 200 USP units of oxytocin. Fertilization rate, which was determined 72 h after AI, was greater (P < 0.05) after laparoscopic than after transcervical/cervical AI (92.5 vs 28%), but oxytocin treatment did not affect fertilization rate. The results indicate that exogenous oxytocin did not reduce ovum fertilization rate, but the transcervical AI procedure per se seemed to reduce fertilization rate.

Journal Article↗
PubMed1997

Regulation of uterine immune function during the estrous cycle and in response to infectious bacteria in sheep.

Uterine infections are a major reproductive problem in livestock. We conducted two experiments to investigate factors that may modulate uterine responses to infectious bacteria. In Exp. 1, ewes received intrauterine inoculations of either saline or bacteria (75 x 10(7) cfu of Actinomyces pyogenes and 35 x 10(7) cfu of Escherichia coli) on either d 0 or 7 of the estrous cycle. Vena caval samples containing uteroovarian blood were collected twice daily from 12 h before until 6 d after inoculation. Only ewes inoculated with bacteria on d 7 developed infections. Basal (4.8 vs .4 pmol), lipopolysaccharide-stimulated (14.2 vs 6.1 pmol), and concanavalin A-stimulated (65.8 vs 21.6 pmol) blastogenesis (i.e., [3H]thymidine incorporation) of vena caval lymphocytes was greater (P < or = .002) for ewes inoculated with bacteria or saline on d 0 rather than on d 7. The number (per 100 white blood cells) of lymphocytes was greater (41.3 vs 30.8, P < .001) and that of neutrophils was less (42.5 vs 51.6, P < .001) in ewes inoculated on d 0 rather than d 7. Bacteria increased (P < .05) vena caval PGF(2 alpha) but not PGE2 concentrations. In Exp. 2, two protein fractions (molecular weights of > or = 100 kDa and approximately 12.7 kDa) from chromatography of uterine flushings collected on d 0 or 7, or 18 d after ovariectomy on d 0 or 7, modulated phytohemagglutinin-stimulated blastogenesis; the heavier fraction from d 0 had a stimulatory component, but the major effects of the fractions were inhibitory. The differences in immune function and regulation between d 0 and 7 probably explain how the uterus of follicular phase ewes was able to prevent the development of an infection.

Actinomyces↗