Evaluation of three-ram cohort serving capacity tests as a substitute for individual serving capacity tests.

Alternatives to time-consuming, laborious individual serving capacity tests (ISCT) are needed to classify ram sexual behavior. The objective of study 1 was to evaluate the relationship between the first 3-ram cohort test (COSCT) scores and the mean of 5 ISCT scores. The objective of study 2 was to determine whether 1 or 2 additional COSCT improved the ability to predict ISCT scores. For study 1, rams (n = 69) were assigned to either a COSCT given before or after 9 ISCT. For study 2, rams (n = 127) were given 3 COSCT before or after 6 ISCT. For repeated COSCT, rams were initially grouped at random and subsequently rerandomized so that each ram was grouped with at least 1 different ram for each test. For both studies, the number of ejaculations from COSCT was compared with the mean number of ejaculations across the second through sixth ISCT. A threshold between high- and low-performing rams was defined in each analysis as the mean ISCT scores of sexually active rams. Rams with a mean number of ejaculations in ISCT greater than the threshold were classified as having high sexual activity, whereas rams below the threshold were classified as having low sexual activity. Rams with no ejaculations in ISCT were classified as sexually inactive. Data from studies 1 and 2 were used to evaluate the relationship between the first COSCT and the mean of 5 ISCT scores. Data from multiple COSCT were fit to various models to determine whether the ability to predict ISCT scores was improved with 1 or 2 additional COSCT. The best model for ISCT and COSCT was a piecewise linear regression model. The first COSCT correctly identified all sexually inactive rams in both studies. The first COSCT, however, also classified 56% of low sexually active rams and 18% of high sexually active rams as inactive. Rams had a 71% probability of high sexual activity in ISCT if they were classified as sexually active in the first COSCT. We conclude that a single COSCT is a reliable, albeit more conservative, and efficient alternative to a series of ISCT for characterizing sexual activity of rams. Multiple COSCT can provide some protection against culling rams with high sexual activity (i.e., approximately 50% less with 2 additional COSCT) and still retain most of the efficiency compared with ISCT. It is important to use high-performance rams for breeding because they will approximately double the number of ewes bred and lambs sired compared with low-performance rams if a large number of ewes need to be serviced daily.

The influence of breeding intensity on above- and below-average sexual performance rams in single- and multiple-sire breeding environments.

Two studies were conducted to evaluate the relationship between serving capacity scores and breeding performance of rams. The first study was conducted to determine whether rams with above or below mean serving capacity scores could perform equally in greater and lesser breeding intensity, single-sire mating schemes. The second study was conducted to determine whether rams with above and below mean serving capacity scores could perform equally well when only one or two ewes were in estrus daily in a multiple-sire breeding scheme (two rams/pen). Rams (n=68) were ranked according to average number of ejaculations recorded in serving capacity tests. Sixteen rams with the greatest scores (above-average) and 16 rams with least scores (below-average) were identified for breeding. Half of above-average and half of below-average rams were used in the two studies. For study 1, each ram was individually introduced to 23 estrus-synchronized ewes for 9 d to simulate high breeding intensity. Rams were given a 5-d rest before they were individually introduced to 23-24 naturally cyclic ewes for 17 d (low breeding intensity). For study 2, 16 rams were paired across ram types, and each pair competed for 20 ewes for 18 d (8 pens). For study 1, ewe fertility (ewes lambing/ewes present at lambing) and number of lambs born were greater (P<0.001) for above-average (0.67+/-0.03 and 27.6+/-1.2, respectively) than for below-average rams (0.39+/-0.07 and 15.3+/-2.7) with greater breeding intensity. Ewe fertility and lambs born did not differ for above-average (0.91+/-0.03 and 37.8+/-1.9, respectively) and below-average rams (0.86+/-0.03 and 39.0+/-1.9) with less breeding intensity. For study 2, number of ewes lambing (99+/-8.0 compared with 72+/-13.6; P=0.12) and number of lambs sired (149+/-18.5 compared with 101+/-22.8; P=0.14) did not differ between above- and below-average rams, respectively, in direct competition. Sexual classifications based on serving capacity tests are related to breeding performance of rams in certain breeding environments. When breeding intensity is greater, above-average rams impregnate more ewes and sire more lambs than below-average rams. When only a small number of ewes are in estrus daily, below-average rams for serving capacity scores perform as well as above-average rams in multiple-sire and single-sire breeding environments. We suggest that above-average rams should be used to reduce number of rams required when breeding intensity is greater.

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.

Comparison of cortisol, luteinizing hormone, and testosterone responses to a defined stressor in sexually inactive rams and sexually active female-oriented and male-oriented rams.

The objective of this study was to determine whether the effect of restraint stress on cortisol, LH, and testosterone varied among sexually inactive and sexually active female- and male-oriented rams, to allow differentiation among ram classes. Restraint stress or no stress was imposed on sexually inactive (n = 7) and sexually active female- (n = 17) and male-oriented (n = 6) rams in a 2 x 3 factorial arrangement. Rams were assigned to restraint or control within each classification. Rams were habituated to wearing halters and being tethered in separate pens, permitting visual, vocal, and olfactory contact with adjacent rams for 7 d before treatment. After 1 d of habituation, rams were fitted with jugular catheters that were checked twice daily for patency. For restraint stress, rams were laid on their side with their legs tied for 1 h. For no stress, rams were tethered with halters and leads, but their legs were not tied. On the treatment day, blood was collected at 30-min intervals for 3 h followed by 15-min intervals for 1 h before restraint, during 1-h restraint, and for 1 h after liberation from restraint. Then blood was collected at 30-min intervals for an additional 2 h. Blood was collected from controls at similar intervals. Control rams were isolated from stressed rams. Cortisol, LH, and testosterone were measured using RIA. Mixed model analyses with repeated measures were used on transformed data. Average prestress data were used as a covariate. Cortisol increased (P < 0.01) within 15 min after restraint and remained increased until 1.5 h after liberation from 1-h of restraint stress. In contrast, in controls cortisol remained unchanged at 5 ng/ mL. Cortisol did not differ over time among ram classes, and the treatment x ram class x time interaction was not significant. For LH and testosterone, the ram class x time interactions appeared to compromise the ability to identify differences in these hormones, indicating that they were not good endocrine candidates for methods of classifying rams. In conclusion, restraint stress increased cortisol in sexually inactive and sexually active female- and male-oriented rams alike, thus not providing a method to differentiate among ram classes.

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.

Testosterone and luteinizing hormone responses to naloxone help predict sexual performance in rams.

The first objective of this study was to determine whether LH and testosterone respond differently to a naloxone injection in relation to varying sexual performance in rams. If differences occurred, the second objective was to determine whether differences would predict variation in sexual performance. From a group of 1.5- to 3-yr-old rams, 20 sexually active and 39 sexually inactive rams were selected based on previously observed sexual behavior with estrual ewes. Each ram was exposed to three estrual ewes for 18 30-min sexual performance tests, and those found to be inactive were given two 30-min sexual partner preference tests. The final distribution was 28 sexually active, 22 inactive, and nine male-oriented rams. Rams were treated with 1.5 mg of naloxone/kg BW in December of Year 1 and again with either 0.75 or 1.5 mg of naloxone/kg BW in November of Year 2. Plasma concentrations of LH and testosterone were evaluated with mixed model analyses for repeated measures separately for each year to coincide with logistic procedures for modeling the probability that rams were sexually active. For Year 1, a sexual activity x age x time interaction for LH after naloxone was observed (P < 0.03). For testosterone, there was a sexual activity x time interaction (P < 0.03), with a similar, early increase for sexually active female-and male-oriented rams compared with a delayed, minimal increase for inactive rams. For Year 2, when all rams were over 2.5 yr of age, a sexual activity x time interaction for both LH and testosterone (P < 0.02) seemed more related to an earlier increase of both hormones for sexually active rams than the increase observed for inactive rams. In addition, sexually active rams had a greater increase in testosterone than inactive rams. No significant difference was observed between 0.75 and 1.50 mg of naloxone/kg BW. Testosterone and LH were used as explanatory variables and sexual activity was used as the response variable in logistic procedures. In Year 1, greatest prediction accuracy was 73.5% using testosterone at 60 min after naloxone injection. In Year 2, the greatest prediction accuracy was 85% using LH at 15 min multiplied by testosterone at 60 min after naloxone. Test repeatability for both years on the same rams was 76%. In conclusion, pattern and magnitude of naloxone-induced changes in endocrine function may facilitate identification of sexually active and inactive rams during the breeding season. Prediction accuracy of the naloxone-based test was 69 to 85%.

Use of naloxone challenge to predict sexual performance of young rams.

The possibility of developing a hormone-based test to predict libido was evaluated using the response of LH and testosterone to naloxone. This test has been used to identify sexually active and inactive mature rams during the breeding season. The objective of this study was to determine whether the blood test could be used to detect differences in sexual activity of early postpubertal (29 +/- 0.1 wk) rams during the breeding season in November and again at 70 +/- 0.1 wk of age in August before the next breeding season. Rams were classed as sexually active or inactive using serving capacity tests (8 30-min observation periods to record sexual behaviors [mounts and ejaculations] of each ram individually exposed to three ewes in estrus) after the naloxone challenges. Naloxone (0.75 mg/kg of BW) was injected i.v. into 38 white-faced crossbred, 16 Polypay, and 49 Targhee rams. Blood samples were collected at 15-min intervals for 1 h before and 2 h after naloxone to measure LH and testosterone. Separate mixed-model analyses for repeated measures were used to analyze data for the same rams at 29 and 70 wk of age. Logistic regression procedures were used to model probabilities that rams were correctly predicted to be sexually active. A breed-type x sexual activity x time interaction for LH was observed (P < 0.05) after naloxone injection in 29-wk-old rams. At 70 wk of age, a breed-type x time interaction was detected (P < 0.001) for LH response to naloxone, but LH did not differ by sexual activity. At 29 wk of age, a breed-type x time interaction for testosterone response after naloxone was detected (P < 0.001), and at 70 wk of age, a sexual activity x time interaction was detected (P < 0.05) for testosterone after naloxone. Sexually active and inactive rams were not predicted accurately at 29 wk of age and were predicted 69 and 29% of the time for sexually active and inactive rams, respectively, at 70 wk of age. In conclusion, breed type at 29 and 70 wk of age can influence the naloxone challenge test, but the test cannot be used to discriminate between sexually active and inactive rams at 29 wk of age during the breeding season or at 70 wk of age immediately before the breeding season.

Ram mating behavior after long-term selection for reproductive rate in Rambouillet ewes.

Mating behavior is known to be heritable in several species, but it is not known if selection schemes for ewe reproductive traits affect mating behavior of rams. Therefore, our objectives were to determine if divergent (high vs low index) selection of a female reproductive trait (lambs born divided by age of ewe minus one) affects mating behavior patterns of male offspring during training to service an artificial vagina (AV) in a less than natural setting (Exp. 1) or during serving capacity tests (SCT) in simulated natural mating conditions (Exp. 2). The method for evaluating male mating behavior was a series of serving capacity tests to estimate sexual performance. For Exp. 1, five, 30-min observations were conducted to assess the mating behavior of 29 rams (22 mo old) being trained to service an AV. In Exp. 1, percentages of rams mounting (73 vs 36%) and ejaculating (67 vs 29%) differed (P < 0.05) between rams from high and low Rambouillet ewe selection lines, respectively. In Exp. 2, ram classification consisted of exposing each 22- to 24-mo-old ram (n = 48) to three unrestrained ewes in estrus for 18, 30-min tests. Any ram that had not mounted or ejaculated during SCT was evaluated for sexual orientation in a 30-min preference test in which each ram had access to both restrained estrual ewes and restrained rams. In Exp. 2, there was no difference (P > 0.2) in percentages of rams mounting (92 vs 78%) and ejaculating (88 vs 74%) between rams from high and low Rambouillet ewe selection lines, respectively. There was no difference (P > 0.33) between high and low ewe selection line rams for mounts or ejaculations for 18 SCT. Only one male-oriented ram was identified, which was from the high line. In Exp. 2, six SCT were necessary to obtain 95% reliability in sexual performance scores. Results of these studies indicate that long-term selection of ewes for litter size did not alter ram sexual performance after SCT under simulated natural mating conditions or affect incidence of male-oriented rams. Mating behavior observed during training to service an AV was lower for rams from ewes of the low selection line than for rams from ewes of the high selection line. We concluded that long-term selection for reproductive rate in ewes did not alter mating behavior patterns of male offspring; however, test conditions, such as restrained and unrestrained ewes and number of serving capacity tests, may affect conclusions in studies that evaluate sexual performance of rams.

Heritability and repeatability of sexual performance scores of rams.

Sexual performance has been subjectively measured with a libido test during screening of rams before public sale and breeding at the U.S. Sheep Experiment Station from 1990 to 2000. The objective of this study was to determine whether sexual performance was genetically influenced. Sexual performance scores ranged from 1 to 6 with scores increasing from sexually inactive to highly sexually active in the presence of estrous ewes. The overall average score was 3.5+/-0.02. Records from four breeds (Columbia, n = 807; Polypay, n = 1,668; Rambouillet, n = 1,208; and Targhee, n = 1,002) were combined into one analysis because breeds had similar phenotypic variances. Total number of records was 4,685, which included a second sexual performance test on 1,212 rams in the following year. Variance components were estimated using a REML algorithm. Fixed effects were breed of ram, selection line within breed, and year by breed. A permanent environmental effect for ram was included to account for repeated observations on individual animals. Age and weight of the rams at time of the libido test were linear covariates and were breed-specific. Adjusted means for sexual performance scores did not differ among breeds (P > 0.05). Age was a significant effect (P < 0.01), with sexual performance score increasing 0.05 units for each month of age. The additive genetic variance was estimated as 0.54. The estimate of variance due to ram permanent environmental effects was 1.19. The residual variance was estimated to be 0.67. The heritability estimate was moderate (0.22+/-0.04) and repeatability was high (0.72). These results imply that one screening for sexual performance provides a reliable measure of sexual performance and that favorable response to selection for ram serving capacity may be expected.

Influence of classification levels of ram sexual activity on spring breeding ewes.

Ram lambs (7-8 months old) and mature rams (19-20 months old) were used to evaluate the effect of classification levels of male sexual performance on reproductive performance of ewes during spring breeding. In Exp. 1, sexually active ram lambs with high (1.8+/-0.3; n=5) and low (0.9+/-0.2; n=5) sexual performance scores (HP and LP; mean+/-S.E.M.) were used in single sire breeding pens. Ewes (n=305) were stratified by age and assigned to 10 pens for 34 days starting in late March. For Exps. 2 and 3, two replicates were conducted for 2 years with sexually active mature rams in a single sire mating scheme. For Exp. 2, HP rams (n=5) averaged 3.6+/-0.2 ejaculations and LP rams (n=7) 1.8+/-0.2 ejaculations for sexual performance scores based on nine, 30 min serving capacity tests (SCT). Polypay ewes (n=152 to 153 per year) were stratified by age and assigned to pens each year for 34-38 days starting in late March for years 1 and 2. For Exp. 3, HP rams (n=6) averaged 3.7+/-0.1 ejaculations and LP rams (n=10) 2.3+/-0.1 ejaculations for sexual performance scores based on 18, 30 min SCT. Polypay ewes (n=229 in year 3 and n=244 in year 4) were stratified by age and assigned to pens each year for 34 days starting in late March. In Exp. 1, lambing rates for ewes bred to HP versus LP ram lambs did not differ (65.8 versus 53.0; P=0.20). Prolificacy tended (P=0.06) to be increased by 0.1 lambs in ewes bred by LP ram lambs. Total number of lambs born per ewe present at lambing, and lambing distribution were not altered by HP and LP ram lambs. In Exp. 2, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (58.1 versus 60.1; P=0.78). In Exp. 3, lambing rates for fall-lambing ewes bred to mature HP or LP rams did not differ (74.3 versus 69.0; P=0.35). There was no difference (P>0.10) between years for Exp. 2 or Exp. 3, and mature HP and LP rams did not affect the other reproductive variables monitored. Analyses of the combined data for Exps. 2 and 3 indicated only a year difference (P<0.001) in lambing rates and total lambs born. Present studies indicate that different sexual performance classifications for ram lambs and mature rams did not alter lambing rates or distribution of lambing of Polypays bred in late March to April. These results indicate that HP and LP, sexually active, Polypay rams and ram lambs with average to high quality semen can provide a source of rams for spring breeding Polypays in ambient conditions and that there was no advantage to using HP over sexually active LP ram lambs or rams.

Expression of the antiviral protein Mx in peripheral blood mononuclear cells of pregnant and bred, non-pregnant ewes.

Interferon-tau (IFN tau) acts locally on the endometrium to suppress estrogen and oxytocin receptor expression and block luteolysis in ruminants. Systemic administration of conceptus homogenates or recombinant ovine IFN tau does not block luteolysis or enhance pregnancy rates in sheep or cattle, respectively. However, IFN tau up-regulates expression of the antiviral protein Mx throughout the entire uterine wall during early pregnancy. These studies determined if conceptus-derived IFN tau also up-regulates Mx expression in components of the circulating immune system that migrate through the endometrial wall. In experiment one, peripheral blood mononuclear cells (PBMC) were isolated from ewes at D26 post-artificial insemination (AI) and Mx mRNA levels examined by Northern and slot-blot hybridization. Pregnancy resulted in a two-fold increase in Mx mRNA levels compared to bred, non-pregnant ewes at D26. In experiment two, PBMC were isolated from ewes at AI, and every three days from D9 to D30. Results showed a four-fold increase in Mx mRNA levels in PBMC from pregnant versus bred, non-pregnant ewes at D15. Increased Mx mRNA, which remained elevated through D30, was accompanied by increased levels of Mx protein. These results show that pregnancy recognition signaling rapidly induces Mx gene expression in PBMC, and are the first to suggest that IFN tau activates gene expression in components of the circulating immune system.

Fos-like immunoreactivity in brain regions of domestic rams following exposure to rams or ewes.

Limbic and basal forebrain-hypothalamic regions from male sheep differing in sexual performance were quantified for fos-like immunoreactivity. Rams classified as high-sexually performing (HP), low-sexually performing (LP), and male-oriented (MO) received noncontact sensory stimulation from either ewes in estrus (HP, n=5; LP, n=4; MO, n=4) or other males (HP, n=5; LP, n=4; MO, n=5) for a 4-h period on each of 3 consecutive days. Following exposure to stimulus animals on the third day, rams were euthanized and their brains were perfused with a 1% paraformaldehyde/1.5% glutaraldehyde solution and sections were analyzed for fos-like immunoreactivity. Brain regions analyzed were the medial amygdala (meAMY), medial preoptic area (mPOA), bed nucleus of the stria terminalis (BNST), and ventromedial hypothalamic nucleus (VMH). Fos-like immunoreactivity differed between groups in the mPOA and BNST but not in the meAMY or VMH. LP rams exposed to estrous ewes had more (P<.05) neurons staining positive for fos and fos-related antigens (FRA) in the mPOA and BNST than LP rams exposed to other rams or MO rams exposed to either estrous ewes or other rams. Numbers of neurons staining positive for FRA in the mPOA and BNST of LP rams exposed to estrous ewes, however, were not different (P>.05) from HP rams exposed to either estrous ewes or other rams. The similar fos-like immunoreactivity in areas important for the display of sexual behavior in HP and LP rams may reflect similar sensory input in these two groups of rams; however, LP rams, in contrast to HP rams, do not appear to respond similarly to the same sensory stimulus.

Reproductive performance of ewe lambs from ewes from different selection practices with or without induced estrus.

Three groups of ewe lambs born in May (experiment 1; n=211) or April (experiment 2; n=174) were used to evaluate the effects of selection line and induction of estrus on pregnancy rate. Experiment 1 was a single factor experiment with induction of estrus as the main effect. In early December, May-born Targhee (n=82) and Rambouillet x Targhee (n=129) ewes were randomly assigned within body weight to one of two treatment groups: control or induction of estrus. Experiment 2 was designed in a 2x2 factorial array with the main effects of induction of estrus or selection line. In early November, April-born Targhee lambs (n=174) from two distinct selection lines were either treated as controls or received an estrus induction treatment. The two lines included an unselected control line of randomly bred ewes and a line that had been selected since 1976, based on the weight of lamb weaned. Ewes from each line were randomly assigned within body weight to one of the treatment groups. In experiments 1 and 2, estrus was induced using MAP pessaries. Pessaries were inserted for 12 days. At the time of pessary removal, ewe lambs received 400 IU eCG i.m. All ewe lambs were bred in multi-sire pens. Pregnancy rate and fetal numbers were determined either by lambing data or real-time ultrasound. Body weight, lambing date and fetal numbers were analyzed by GLM, and remaining variables were analyzed by CATMOD. For experiment 1, estrus induction increased (P<0.01) pregnancy rates (61 versus 31%) and number of fetuses estimated by real-time ultrasound (79 versus 35%) compared to control ewe lambs. Pregnancy rate and fetal number were increased (P<0.01) for the 1st year compared to the 2nd year. For experiment 2, estrus induction tended to increase (P<0.07) pregnancy rate, and pregnancy rate differed (P<0.01) between selection lines. Estrus induction increased (P<0.05) fetal numbers (0.96) compared to controls (0.77). Fetal numbers were greater (P<0.01) for the selected line (1.06) compared to random bred controls (0.67). Average date of lambing was earlier in both experiments for the estrus-induced ewe lambs compared to controls. These results indicate that induction of estrus can be recommended if increased reproduction is desired for ewe lambs.

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.

Low-sexually performing rams but not male-oriented rams can be discriminated by cell size in the amygdala and preoptic area: a morphometric study.

Brain regions of male sheep behaviorally classified as high-sexually performing (n=10), low-sexually performing (n=8) or male-oriented (n=9) were examined to determine if differences in reproductive behavior were associated with differences in density or sizes of neurons. High-sexually performing rams actively mounted estrous ewes, low-sexually performing rams failed to mount or had long latencies to mounting estrous ewes, and male-oriented rams mounted other rams in preference to ewes in estrus. Cell densities and sizes were quantified in Nissl stained sections through the medial amygdala (meAMY), preoptic area (POA), bed nucleus of the stria terminalis (BNST), ventromedial hypothalamic nucleus (VMH), lateral geniculate nucleus (LG) and medial geniculate nucleus (MG). Multivariate discriminant analysis based on soma sizes within nuclei of known importance for reproductive behavior and/or gonadotropin release (meAMY, POA, BNST and VMH) discriminated (Wilks Lambda P<0.05) low-performing rams from high-performing and male-oriented rams, but did not discriminate (Wilks Lambda P=0.14) between high-performing and male-oriented rams. Cell size in the parvocellular and magnocellular layers of the LG along with cells of the MG, structures without a specific role in reproduction, did not discriminate any of the three behaviorally defined groups of rams (Wilks Lambda P=0.57). Density of cells present in structures important for the display of reproductive behavior (POA, meAMY, BNST) and/or gonadotropin release (POA, VMH) had no discriminating power nor did density of cells in structures important for the processing of visual (LG) or auditory (MG) stimuli. In conclusion, significant differences in sizes of cells located within nuclei that are specifically important for the display of male reproductive behavior were found in low-sexually performing rams compared to high-sexually performing and male-oriented rams. These differences may result from neuron development in utero or occur later as a consequence of endocrine factors or behavioral experience. Neuronal cell size is a critical variable that determines excitability to synaptic inputs because cell surface area varies exponentially with cell diameter. Relatively small differences in neuron diameter could relate to functionally important differences in neuronal excitability.

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.

Behavior and endocrine changes in high-performing, low-performing, and male-oriented domestic rams following exposure to rams and ewes in estrus when copulation is precluded.

High-performing, low-performing, and male-oriented rams were used to investigate behavior and neuroendocrine correlates of sexual interest and discrimination. Treatment consisted of visual and olfactory contact with stimulus animals through a woven wire fence, which inhibited copulation (either ewes in estrus or other rams), for 4 h on each of three consecutive days. Before exposure to stimulus animals on d 1 and during the final 1 h of exposure on d 2, blood samples were collected every 15 min for 1 h to determine concentrations of LH and testosterone. During exposure to stimulus animals, rams were continuously observed and investigatory behaviors were recorded. There was no day effect for any behavior. Groups of rams differed (P < .05) in amounts of behaviors exhibited, but behaviors were not influenced by sex of stimulus animals. High-performing rams exhibited more (P < .05) investigatory behaviors toward stimulus animals than low-performing or male-oriented rams. Plasma concentrations of LH increased (P < .05) in high-performing rams following exposure to estrous ewes, but not following exposure to rams. In low-performing and male-orientated rams, concentrations of LH were unchanged regardless of sex of the stimulus animal. Change in plasma concentrations of testosterone from pre- to posttreatment did not differ between high-performing, low-performing, and male-oriented rams. However, low-performing rams exhibited an increase (P < .05) in plasma concentrations of testosterone following exposure to rams. In conclusion, high-performing rams exhibit a high degree of investigatory behaviors toward estrous ewes and other rams. High-performing rams seem to discriminate sex of stimulus animals and exhibit a neuroendocrine response (i.e., increased plasma LH) only when exposed to ewes in estrus. The sensory signals provided by estrous females are either not detected by low-performing or male-oriented rams or are not sufficiently provocative to elicit further investigation by these rams.

Sexual behaviour of rams: male orientation and its endocrine correlates.

The components of heterosexual behaviour in rams are reviewed as a basis for understanding partner preference behaviour. A small percentage of rams will not mate with oestrous females and if given a choice will display courtship behaviour towards another ram in preference to a female. Some of the endocrine profiles of these male-oriented rams differ from those of heterosexual controls. These differences include reduced serum concentrations of testosterone, oestradiol and oestrone, reduced capacity to produce testosterone in vitro, and reduced capacity to aromatize androgens in the preoptic-anterior hypothalamus of the brain. Our observation that aromatase activity is significantly lower in the preoptic-anterior hypothalamic area of male-oriented rams than in female-oriented rams may indicate an important neurochemical link to sexual behaviour that should be investigated. The defect in steroid hormone production by the adult testes of the male-oriented ram may represent a defect that can be traced to the fetal testes. If this contention is correct, partner preference behaviour of rams may also be traceable to fetal development and represent a phenomenon of sexual differentiation.

Effect of selection for lifetime production of lamb weaned on hormonal factors that affect growth in Targhee ewes and lambs.

Targhee ewes (n = 22, average age 5 yr) rearing twin lambs were used to investigate serum growth hormone (GH), IGF-I, triiodothyronine (T3), thyroxine (T4), and prolactin (PRL; ewes only) concentrations associated with selection pressure for lifetime production of kilograms of lamb weaned per ewe. Period 1 started on d 4 after birth and was conducted in confinement. Periods 2, 3, and 4 were conducted on fenced intermountain sagebrush-bunchgrass range starting at an average of 49, 84, and 112 d after birth, respectively. Blood samples were collected hourly for 6 h on d 4, 11, 18, 25, 49, 84, and 112 after lambing. Ewe models included the fixed effect for line, with ewe age, lambing date, and starting weight nested within line as potential covariables. Lamb models included fixed effects for line, sex, and sibling sex, with lamb age and birth weight nested within line tested as possible covariables. Growth hormone concentrations were greater (P = .06) for selected than for control ewes, but lamb GH concentrations did not differ (P = .90) between selected and control lambs. Ewe prolactin concentration tended (P = .13) to be greater for control than selected ewes. Ewe and lamb IGF-I did not differ (P > .55) between selected and control ewes and lambs. Ewe T3 and T4 concentrations did not differ (P > .19) between selected and control ewes; however, lamb T3 and T4 concentrations were greater (P < .01) for control than for selected lambs. Increased GH concentration in selected ewes seemed to be associated with greater milk production. Differences in selected and control lamb T3 and T4 concentrations could be an indicator of receptor sensitivity, metabolic activity, or seasonal reproductive transitions in ewe lambs.

Influence of simulated photoperiod alteration and induced estrus on reproductive performance of spring-born Columbia and Targhee ewe lambs.

The objective of this study was to test the hypothesis that a combination of induced estrus (IE) and melatonin (MEL) would increase reproductive performance in breeds characterized by late maturity more than either IE or MEL alone. Spring-born Columbia (C; n = 161; 188 to 222 d range of age at breeding; 49 to 80 kg range of BW at breeding) and Targhee (T; n = 166; 183 to 210 d range of age at breeding; 40 to 68 kg range of BW at breeding) ewe lambs were stratified randomly by breed to treatments: ambient controls (AC); IE; MEL; and MEL+IE. Melatonin (18 mg of Regulin) was implanted on September 7. Estrus induction included medroxyprogesterone acetate (MAP) pessaries inserted for 12 d and pregnant mare's serum gonadotropin (PMSG; 500 IU i.m.) at time of MAP removal. Ewe lambs were placed into two pens with five rams per pen by breed. Targhee ewes lambed at an increased rate with IE vs no IE (84.5 vs 67.9%; P < .01) and tended to lamb at an increased rate with MEL vs no MEL (82.5 vs 70.6%; P = .07). The 92.7% lambing rate observed in the Targhee ewe lambs for the combined treatment (MEL+IE) was slightly higher than an additive effect but the interaction between IE and MEL was not significant (P = .24). By contrast, none of the treatments altered (P = .5) lambing rate, number of lambs born, or number born alive for Columbia ewe lambs.(ABSTRACT TRUNCATED AT 250 WORDS)