KAP1．3基因与绵羊产毛性能的相关分析

本实验采用PCR—SSCP的标记技术,对羊毛纤维组成蛋白中的KAP1.3的部分序列进行多态性的研究。结果表明,在引物1中,BB基因型的产毛量性状极显著高于其他基因型（p〈0．01）,在拉伸长度性状上,AB基因型与BB基因型差异极显著（p〈0．01）,AA基因型与BB基因型差异显著（0．01〈p〈0．05）。在引物2中,各基因型不存在显著差异。     

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.     

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.     

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.     

Assessment of Meiotic Spindle Configuration and Post‐Warming Bovine Oocyte Viability Using Polarized Light Microscopy

The objectives of this study were to assess the efficiency of polarized light microscopy (PLM) in detecting microtubule‐polymerized protein in in vitro‐matured bovine oocytes; to examine its effects on oocyte developmental competence; and to assess the meiotic spindle of in vitro‐matured oocytes after vitrification/warming and further assessment of oocyte developmental competence. In the first experiment, the presence of microtubule‐polymerized protein (MPP) was confirmed as a positive PLM signal detected in 99.1% of analysed oocytes (n = 115), which strongly correlated (r = 1; p < 0.0001) with the presence of MPP as confirmed by immunostaining. In the second experiment, oocytes (n = 651) were exposed or not (controls) to PLM for 10 min and then fertilized and cultured in vitro. Oocytes exposed to PLM did not significantly differ from controls with regard to cleavage, total blastocyst and expanded blastocyst rates and cell numbers. In the third experiment, meiotic spindles were detected in 145 of 182 oocytes (79.6%) following vitrification and warming. Interestingly, after parthenogenetic activation and in vitro culture, oocytes that displayed a positive PLM signal PLM(+) differed significantly from PLM(?) in cleavage and Day 8 blastocyst rates. These results suggest that polarized light microscopy is an efficient system to detect microtubule‐polymerized protein in in vitro‐matured bovine oocytes and does not exert detrimental effects on bovine oocyte developmental competence. Moreover, PLM could be used as a tool to assess post‐warming viability in vitrified bovine oocytes.     

Gastric squamous cell carcinoma in three horses

Gastric squamous cell carcinoma was diagnosed in three horses. Clinical signs observed in all cases were weight loss, anorexia and lethargy. Respiratory signs were prominent in one case. All three horses had depressed albumin and elevated globulin and fibrinogen concentrations. Two horses were mildly anaemic. Inflammatory exudates were present in peritoneal cavities in all cases, and cytological evaluation provided a positive diagnosis of squamous cell carcinoma in two cases. Pleural fluid samples taken from two cases were also classified as inflammatory exudates, but no neoplastic cells were detected on initial examination. In all cases the neoplasms had arisen from the oesophageal region of the stomach, and had metastised throughout the abdomen. Two cases had metastatic lesions within the pleural cavity.     

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.