Extraction methods in bovine obstetrics: comparison of the demanded energy and importance of calf and traction method in the variance of force and energy

In a previous study we could demonstrate that in terms of the required forces alternate traction is preferable to simultaneous traction for the obstetrical delivery of calves. The aims of this study were to examine the required energies for delivery of calves and to compare the effects of the factors of calf and traction method on the forces and energies required for the delivery. In a biomechanical in vitro model 12 stillborn Holstein-Friesian calves were pulled through the pelvic specimen of a cow at a controlled speed using two electric motors. Traction was applied simultaneously (ST) to both legs or alternately (AT) to one leg at a time to advance it 10 cm (AT 10) or 5 cm (AT5). Energies on each limb were measured digitally using load cells. The lowest energy for the entrance of the elbows in the pelvis was necessary using AT10 (19.9 ± 7.2 kJ, P < 0.05). In contrast, for the entrance of the chest, AT10 (104.9 ± 24.7 kJ) demanded 9% (P < 0.01) and 16% (P < 0.001) more energy than AT5 (96.7 ± 21.0 kJ) and ST (90.5 ± 24.9 kJ), respectively. Simultaneous traction tended to be better than AT5 (P = 0.09). Variance component estimates revealed that the factor calf contributed the main effect to the emerging forces and required energy. The traction method was responsible for up to 13% of the variance in most of the cases, but it accounted for 42% of the variance regarding the forces on the lower limb as the chest entered the pelvis. Based on these findings, the decision of the clinician whether or not to perform a manual traction should depend mainly on the calf. However, when such a decision has been made, alternate limb traction, 10 cm at a time, should be used until both elbows have entered the pelvis and a simultaneous traction should then be applied to complete extraction of the chest.     

Closed pulled straw vitrification of in vitro-produced and in vivo-produced bovine embryos

The objective of this study was to evaluate the efficiency of the closed pulled straw (CPS) method for cryopreserving in vitro-produced and in vivo-produced bovine (Bos taurus) embryos. Based on the open pulled straw (OPS) protocol, the top end of a CPS was closed by tweezers (heated in a flame) to prevent the cryoprotectant medium containing embryos from contacting the liquid nitrogen. Bovine in vitro or in vivo morulae and early blastocyst embryos were frozen by slow cryopreservation, OPS vitrification, or CPS vitrification. Morphology of postthawed embryos was evaluated, and normal embryos were used for successive culture for 72 h. There were no significant differences between OPS and CPS freezing groups in postthawed in vitro-produced embryos with respect to rates of morphologically normal embryos (mean ± SD, 87.9 ± 5.2% vs. 85.4 ± 4.9%), survival at 24 h (58.0 ± 6.8% vs. 56.3 ± 4.4%), and survival at 72 h (35.2 ± 6.0% vs. 34.9 ± 6.7%). However, both OPS and CPS vitrification resulted in higher postthaw rates of morphologically normal embryo and survival at 24 and 72 h than those of the slow-freezing method (P < 0.05). Similar results were obtained for in vivo-derived embryos. We concluded that CPS vitrification was a feasible method to cryopreserve both in vitro-derived and in vivo-derived bovine embryos. This method not only eliminated the risk of embryo contamination by preventing contact with liquid nitrogen but also retained the advantages of the OPS vitrification method.     

Ovarian antral follicular dynamics in sheep revisited: Comparison among estrous cycles with three or four follicular waves

In this study, the characteristics of ovarian follicular waves and patterns of serum concentrations of follicle-stimulating hormone (FSH), estradiol, and progesterone were compared between cycles with three (n = 9) or four (n = 10) follicular waves in Western White Face (WWF) ewes (Ovis aries). Transrectal ultrasonography and blood sampling were performed daily during one cycle. Estrous cycles were 17.11 ± 0.3 and 17.20 ± 0.2 d long in cycles with three and four waves, respectively (P > 0.05). The first interwave interval and the interval from the emergence of the final wave to the day of ovulation were longer in cycles with three waves compared with those in cycles with four waves (P < 0.05). The growth phase (5.1 ± 0.5 vs. 3.1 ± 0.4 d) and life span (5.67 ± 0.3 vs. 4.3 ± 0.3 d) of the largest follicle growing in the last or ovulatory wave was longer in cycles with three waves compared with that in cycles with four waves (P < 0.05). The maximum diameter of the largest follicle was greater in the first wave and the ovulatory wave compared with that in other waves of the cycle (P < 0.05). The regression phase of the largest follicle growing in the first wave was longer in cycles with three waves compared with that in cycles with four waves (4.44 ± 0.4 vs. 3.4 ± 0.4 d; P < 0.05). The length of the life span, regression phase, and, although not significant in every case, FSH peak concentration and amplitude decreased across the cycle (P < 0.05). We concluded that estrous cycles with three or four follicular waves were confined within the same length of cycle in WWF ewes. In this study, there were no apparent endocrine or follicular characteristics that could explain the regulation of the different number of follicular waves (three vs. four) during cycles of similar length.     

Reproductive performance of repeat breeders in dairy herds

The objectives were to characterize repeat breeding in dairy cows, including reproductive performance and risk factors. Data from 613 Holstein Friesian cows in nine dairy herds across Japan were enrolled. A repeat breeder was defined as a cow that did not become pregnant after three inseminations, despite no clinically detectable reproductive disorders. In contrast, cows that became pregnant within three inseminations were considered to have normal fertility. Of the 613 cows, 87.3% eventually became pregnant after repeated AI (maximum calving to conception interval was 435 d). Mean (±SEM) first AI conception rate, days in milk at first AI, calving to conception interval and service per conception were 38.3%, 82 ± 2 d, 125 ± 3 d, and 2.0 ± 0.1 times, respectively. Normal fertility cows (n = 479) required only 114 ± 3 d to conceive and 1.7 ± 0.1 inseminations per pregnancy, whereas repeat breeders (n = 86) required significantly more days to conceive (211 ± 10) and more inseminations per pregnancy (4.7 ± 0.2). Based on survival analysis, it took 94 d after calving for 50% of normal fertility cows to become pregnant, compared to 155 d for repeat breeders. For repeat breeders, 31.4, 50.0, and 58.1% became pregnant within 210, 300, and 435 d after calving, respectively. The risk factors for repeat breeding were parity (relative risk [RR] = 0.809; P = 0.058), resumption of postpartum ovarian cycles (RR = 1.928; P = 0.009), and days in milk at first AI (RR = 0.991; P = 0.039). In conclusion, repeat breeder dairy cows had very poor reproductive performance. Lower parity, abnormal resumption of postpartum ovarian cycles, and shorter days in milk at first AI were risk factors for repeat breeding.     

Aneuploidy in rabbit males: semen traits and fertility

Chromosomal analyses were performed from peripheral blood samples from 20 adult rabbit males from Line R. This line has been selected for growth rate after weaning through 25 generations. Seminal characteristics and reproductive outcomes from these males were also examined. The chromosomal analysis results showed that one male had an aneuploidy rate of 16% (male A). The aneuploidies found represented both hypo- and hyper- haploidy. Differences between the male A and the contemporary males (males N), in fertility at 12 days post-insemination (44% vs. 66%) and at birth (31% vs. 59%) were observed. The male A also showed a high percentage of pregnancy losses (29% vs. 12%). For seminal characteristics, the percentage of abnormal spermatozoa was statistically different (P < 0.05) between male A and males N (34 ± 4 vs. 18 ± 2), and concentration differed significantly between males, showing the male A reduced fewer spermatozoa than males N (67 ± 23 vs. 172 ± 10, x106/ml). Motility and kinematic parameters revealed no differences between males. Differences between male reproductive performance found in this study could be explained by the effect of aneuploidy on spermatogenesis and its deleterious effect on male reproduction.     

Physiologic and nonphysiologic effects of exogenous prostaglandin F2α on reproductive hormones in mares

Responses to intravenous treatment of mares with prostaglandin F2α (PGF) 8 d after ovulation were studied in three groups (n = 4/group): control (no treatment), bolus (single treatment with 2.5 mg PGF), and infusion (0.1 mg PGF during 2 h). Infusion resulted in a 13,14-dihydro-15-keto-PGF2α (PGFM) concentration (559 ± 44 pg/mL) that was not different from the mean concentration for the major portion of a natural PGFM pulse associated with luteolysis (569 ± 45 pg/mL; n = 5). Progesterone in the bolus group increased (P < 0.03) between 0 (17.8 ± 3.5 ng/mL) and 2 min (25.3 ± 4.8 ng/mL), peaked at 10 min (28.5 ± 4.6 ng/mL), and then decreased. In the infusion group, progesterone decreased (P < 0.05) during 1 min (17.2 ± 1.3 ng/mL) to 15 min (13.5 ± 1.5 ng/mL) after the beginning of infusion and decreased (P < 0.05) similarly to the bolus group during 2 to 12 h; concentrations were lower (P < 0.05) at each hour than in controls. Interval between ovulations was shorter (P < 0.05) in the bolus (19.3 ± 2.0 d) and infusion (18.8 ± 2.1 d) groups than in controls (24.3 ± 1.3 d). Concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and cortisol increased (P < 0.05) within 10 min in the bolus group but did not change in the infusion group. Results supported the hypothesis that increases in progesterone, FSH, LH, and cortisol after a bolus luteolytic PGF treatment are nonphysiologic. Past conclusions on the nature of the luteolytic mechanism are problematic if based on responses to treatment with a single luteolytic bolus of PGF.     

Effects of bovine Herpesvirus Type 5 on development of in vitro-produced bovine embryos

Bovine (Bos indicus) herpesviruses have been associated with reproductive disease. Type 1, the most studied species, is best known for its reproductive and respiratory effects. Type 5 (BoHV-5) has been detected in bull semen and aborted fetuses but not in oocytes and embryos. This study consisted of three experiments that evaluated (1) BoHV-5-infected oocytes matured in medium with fetal bovine serum (BoHV-FBS) or polyvinyl alcohol (BoHV-PVA) and fertilized by noninfected sperm; (2) noninfected oocytes fertilized by BoHV-5-infected sperm; and (3) infection of presumptive zygotes by BoHV-5. Each treatment involved nine drops of 15 to 20 oocytes. Infection with BoHV-5 was detected by polymerase chain reaction and in situ hybridization assay, and fertilization capacity and embryonic development were assessed using in vitro culture. Experimentally induced infection was obtained in all experiments, and vertical transmission of BoHV-5 by gametes was confirmed. The cleavage rate was reduced (P = 0.0201) in BoHV-FBS (80.4 ± 8.9%; mean ± SD) compared with that of noninfected oocytes (89.9 ± 6.5%); neither differed from BoHV-PVA (87.3 ± 7.1%), and the resulting embryo production rate was not significantly different among groups. Rates of cleavage (87.5 ± 7.5% vs. 92.2 ± 5.5%, control vs. infected) and development of embryos (41.7 ± 9.9% vs. 44.3 ± 7.7% to morula/blastocyst/expanded blastocyst [M/B/EB] and 39.6 ± 10.3% vs. 40.8 ± 9.2% to blastocyst/expanded blastocyst/hatching blastocyst [B/EB/HB] stages) were not compromised by infected sperm (P = 0.1462, P = 0.5402, and P = 0.8074, respectively). However, presumptive zygotes directly infected 1 d after fertilization produced a lower number (P = 0.0140 to M/B/EB and P = 0.002 to B/EB/HB stages) of in vitro-produced embryos (31.6 ± 4.6 vs. 25.0 ± 5.5 and 31.6 ± 4.6 vs. 20.2 ± 5.4; control vs. infected). In conclusion, BoHV-5 infected gametes and was transmissible to the embryo during in vitro development. As zygotes infected 1 d after fertilization had compromised development, BoHV-5 has the potential to be a pathogen with economic consequences.     

Equine chorionic gonadotropin and gonadotropin-releasing hormone enhance fertility in a norgestomet-based, timed artificial insemination protocol in suckled Nelore (Bos indicus) cows

Two experiments were conducted to investigate the effects of equine chorionic gonadotropin (eCG) at progestin removal and gonadotropin-releasing hormone (GnRH) at timed artificial insemination (TAI) on ovarian follicular dynamics (Experiment 1) and pregnancy rates (Experiment 2) in suckled Nelore (Bos indicus) cows. Both experiments were 2 × 2 factorials (eCG or No eCG, and GnRH or No GnRH), with identical treatments. In Experiment 1, 50 anestrous cows, 134.5 ± 2.3 d postpartum, received a 3 mg norgestomet ear implant sc, plus 3 mg norgestomet and 5 mg estradiol valerate im on Day 0. The implant was removed on Day 9, with TAI 54 h later. Cows received 400 IU eCG or no further treatment on Day 9 and GnRH (100 μg gonadorelin) or no further treatment at TAI. Treatment with eCG increased the growth rate of the largest follicle from Days 9 to 11 (means ± SEM, 1.53 ± 0.1 vs. 0.48 ± 0.1 mm/d; P < 0.0001), its diameter on Day 11 (11.4 ± 0.6 vs. 9.3 ± 0.7 mm; P = 0.03), as well as ovulation rate (80.8% vs. 50.0%, P = 0.02), whereas GnRH improved the synchrony of ovulation (72.0 ± 1.1 vs. 71.1 ± 2.0 h). In Experiment 2 (n = 599 cows, 40 to 120 d postpartum), pregnancy rates differed (P = 0.004) among groups (27.6%, 40.1%, 47.7%, and 55.7% for Control, GnRH, eCG, and eCG + GnRH groups). Both eCG and GnRH improved pregnancy rates (51.7% vs. 33.8%, P = 0.002; and 48.0% vs 37.6%, P = 0.02, respectively), although their effects were not additive (no significant interaction). In conclusion, eCG at norgestomet implant removal increased the growth rate of the largest follicle (LF) from implant removal to TAI, the diameter of the LF at TAI, and rates of ovulation and pregnancy rates. Furthermore, GnRH at TAI improved the synchrony of ovulations and pregnancy rates in postpartum Nelore cows treated with a norgestomet-based TAI protocol.     

The effect of hormone treatments (hCG and cloprostenol) and season on the incidence of hemorrhagic anovulatory follicles in the mare: A field study

The association between use of hormone treatments to induce estrus and ovulation and the incidence of hemorrhagic anovulatory follicles (HAFs) was studied in a mixed population of mares (Equus caballus) during two breeding seasons in a commercial breeding clinic. Mares treated with cloprostenol (CLO) were more likely to develop HAFs than were mares with spontaneous cycles (P < 0.001) or those treated with human chorionic gonadotropin alone (P = 0.08). There was no significant effect of season on the incidence of HAFs. The mean (±SEM) interval from CLO treatment to beginning of HAF development was 6.1 ± 0.5 d. Age of mares with HAF cycles was not different (12 ± 1.3 yr; P > 0.05) from that of mares with ovulatory cycles (10.5 ± 1.5 yr).     

Association between evaluation of the reproductive tract by various diagnostic tests and restoration of ovarian cyclicity in high-producing dairy cows

The uterine condition of clinically normal postpartum Holstein-Friesian dairy cows (n = 45) was evaluated once weekly (Weeks 3 to 7) by endometrial cytology, vaginal mucus collection device (VMCD), vaginoscopy, and ultrasonography to establish a relationship with postpartum resumption of ovulatory cycles. The time of first detection of the corpus luteum (CL) by ultrasonography and plasma progesterone concentration ≥1 ng/mL was recorded. By 49 d postpartum, 78% of the cows (n = 35) had resumed ovarian function (CL group), whereas the remainder (n = 10) had no CL (NCL group). There was a positive correlation between VMCD score and presence of fluid in the uterus in cows with a CL (P < 0.01) during Week 3 postpartum but no significant correlation in cows without a CL. Percentage of polymorphonuclear leukocytes (PMN%) was higher in the NCL group (mean ± SEM, 24.6 ± 9.4%) than in the CL group (11.7 ± 2.2%) during Week 5 postpartum (P < 0.05). The PMN% (4.5 ± 6.5%) and VMCD (0.5 ± 0.5) scores during Week 5 in cows ovulating by Day 28 were lower (P < 0.01) than the PMN% (15.0 ± 14.3%) and VMCD (1.1 ± 0.9) scores in those ovulating by Day 49. In conclusion, higher PMN% at 5 wk postpartum was associated with delayed resumption of ovarian cyclicity in high-producing dairy cows.     

Effect of cytoplasmic lipid content on in vitro developmental efficiency of bovine IVP embryos

The objective was to evaluate the effect of cytoplasmic lipid content on the embryonic developmental efficiency of bovine in vitro embryo production (IVP) embryos. Ovaries from Korean native cows (Bos taurus coreanae) were collected from a local abattoir, and cumulus-oocyte complexes (COCs) were recovered from follicles 2 to 8 mm in diameter. The oocytes were divided into three groups, dependent on their cytoplasm color: pale color (PC), brown color (BC), and dark color (DC). The COCs were fertilized using frozen-thawed semen from a single Hanwoo bull. Based on measurement of the cytoplasmic color intensity of oocytes after 22 h of in vitro maturation (IVM), the DC group had lower (P < 0.05) color intensity than that in the BC and PC groups (56.3 ± 2.7, 93.3 ± 5.1, and 123.9 ± 12.0, respectively). Based on MitoTracker Green FM staining, the number of mitochondria in the DC (170.1 ± 31.2) group was significantly higher than that in the BC (137.5 ± 30.8) and PC (105.5 ± 25.3) groups. The cleavage rate in the DC (81.5%) group was also higher than that in the PC (50.4%) group (P < 0.05), as was the development rate to blastocyst stage (18.9% vs. 9.8%). Finally, cell numbers of blastocysts in the DC (150.8 ± 28.0) group were higher (P < 0.05) than that in the BC (107.6 ± 17.8) and PC (80.5 ± 12.3) groups. In conclusion, cytoplasm color was a useful selection parameter for abattoir-derived oocytes destined for IVP.     

Determination of heart rate and heart rate variability in the equine fetus by fetomaternal electrocardiography

Heart rate is an important parameter of fetal well-being. We have analyzed fetal heart rate (HR) and heart rate variability (HRV) by fetomaternal electrocardiography (ECG) in the horse (Equus caballus) from midpregnancy to foaling. It was the aim of the study to detect changes in the regulation of fetal cardiac activity over time and to establish normal values in undisturbed pregnancies. A total of 22 mares were available for the study. Fetomaternal electrocardiography was a reliable technique to detect cardiac signals in fetuses between Day 173 of gestation and foaling. Fetal HR decreased from 115 ± 4 beats/min (Days 170 to 240 of gestation) to 83 ± 3 beats/min (Day 320) to 79 ± 1 beats/min (1 d before foaling; P < 0.001). Mean beat to beat (RR) interval and standard deviation of the RR interval (SDRR) increased (P < 0.001). Gestational age thus affects RR interval and HR in the equine fetus. From Days 270 to 340 of gestation, SDRR increased from 11.4 ± 1.3 msec on Day 270 to 27.8 ± 3.6 msec on Day 340 (P < 0.05), and the root mean square of successive RR differences (RMSSD) tended to increase (P = 0.07), indicating maturation of the fetal autonomous nervous system. For the last 10 d before foaling, fetal HR and HRV remained constant and did not allow predicting the onset of parturition in the horse. Only during the last 30 min before the foal was born, in 4 of 5 fetuses, HR decreased and RR interval increased. Accelerations and decelerations in HR were detectable at all times, but neither their number nor duration changed over time.     

Effect of a GnRH antagonist on GnRH agonist-implanted anestrous bitches

Various combinations of gonadotropin-releasing hormone (GnRH) antagonists and long-acting GnRH agonists have been assessed in several species to prevent the “flare-up” effect that agonists cause on the pituitary-gonadal axis. To determine the effect of a single administration of the GnRH antagonist acyline in anestrous GnRH agonist-implanted domestic bitches, 19 dogs (canis familiaris) were randomly assigned to receive either 10 mg sc deslorelin acetate (DA; n = 6) or DA combined with 330 μg/kg sc acyline within the first 48 h (DA&ACY; n = 13). These bitches were examined daily for detection of posttreatment flare-up, manifested as an estrous response during the month after treatment. In the DA and DA&ACY groups, an estrous response was detected in 6 of 6 and 9 of 12 (P < 0.5) of the bitches, starting 5.3 ± 1.3 and 10.1 ± 1.8 d (mean ± SEM, P = 0.5), respectively, after treatment. Based on serum progesterone concentrations, ovulation occurred in 6 of 6 and 5 of 9 of these bitches (P = 0.1). None of the dogs had any local or systemic side effects related to the treatments. In five DA and six DA&ACY bitches that could be followed up after the trial, interestrus intervals were 385 ± 22.5 and 330 ± 69.1 d, respectively (P > 0.1). It was concluded that the current antagonist protocol prevented initial ovarian stimulation in one quarter of the treated dogs, whereas the stimulation period was postponed and ovulation was inhibited in approximately half of the remainder.     

Embryo production in superovulated Angus cows inseminated four times with sexed-sorted or conventional, frozen-thawed semen

This study tested the hypothesis that four inseminations of commercially frozen sexed semen (≥2.1 × 10⁶ sperm per 0.25-mL straw) in superstimulated embryo donors would yield a percentage and quantity of transferable embryos similar to that achieved with conventional frozen semen. Bos taurus, angus cows (n = 32), stratified by age and body condition, were randomly allocated to receive four inseminations of frozen-thawed semen, either conventional semen (≥15 × 10⁶ sperm/straw; Conventional) or sexed semen (≥2.1 × 10⁶ sperm/straw; Sexed) from one of two AI sires. From 10 to 13 d after estrus, follicle-stimulating hormone (FSH) was given twice-daily, with prostaglandin F_2α given twice on the last day. Cows were inseminated once (1×) at first detected estrus and twice (2×) and once (1×) at 12 and 24 h later, respectively, with nonsurgical embryo recovery 7 d after first detected estrus. The study was repeated 30 d later (switch-back experimental design). The total number of ova per flush was similar between Conventional and Sexed treatments (10.9 ± 1.8 vs. 10.5 ± 1.6), but the number of Grade 1 embryos was greater (P < 0.01) for Conventional (4.3 ± 0.8 vs. 2.3 ± 0.7). Conversely, the mean number of unfertilized ova was greater (P < 0.05) for Sexed (5.6 ± 1.0 vs. 3.0 ± 1.2). There was no significant difference between treatments for numbers of degenerate, Grades 2 or 3, and transferable embryos and no significant differences between bulls in percentage of transferable embryos (44.4% and 46.7%). However, fertilization rates and percentage of transferable embryos were affected (P < 0.05) by period and donor. In conclusion, superstimulated donor cows inseminated four times had fewer Grade 1 embryos and more unfertilized ova with sexed versus conventional semen.     

Evaluation of the ultrasound image attributes of developing ovarian follicles in the four follicular waves of the interovulatory interval in ewes

Computer-assisted quantitative echotextural analysis was applied to ultrasound images of antral follicles in the follicular waves of an interovulatory interval in sheep. The ewe has three or four waves per cycle. Seven healthy, cyclic Western White Face ewes (Ovis aris) underwent daily, transrectal, ovarian ultrasonography for an interovulatory interval. Follicles in the third wave of the ovulatory interval had a longer static phase than that of those in Waves 1 and 2 (P < 0.05). The numeric pixel value for the wall of anovulatory follicles emerging in the third wave of the cycle was significantly higher than that for Waves 1 and 2 at the time of emergence (156.7 ± 8.09, 101.6 ± 3.72, and 116.5 ± 13.93, respectively), and it decreased as follicles in Wave 3 reached maximum follicular diameter (P < 0.05). The numeric pixel value of the antrum in the ovulatory follicles decreased as follicular diameter increased to ≥5 mm in diameter (P < 0.05). The pixel heterogeneity of the follicular antrum in Wave 1 increased from the end of the growth phase to the end of the regression phase for follicles in that wave (P < 0.05). The total area for the wall and antrum of the follicles studied were correlated with follicular diameter in all follicular waves (r = 0.938, P < 0.01 and r = 0.941, P < 0.01 for the wall and antrum, respectively). Changes in image attributes of the follicular wall and antrum indicate potential morphologic and functional differences among antral follicles emerging at different stages of the interovulatory interval in cyclic ewes.     

Effects of different extenders and centrifugation/washing on postthaw microscopic-oxidative stress parameters and fertilizing ability of Angora buck sperm

The main objective of the current study was to evaluate the effects of extender type and centrifugation/washing prior to cryopreservation on the postthaw sperm parameters, lipid peroxidation, and superoxide dismutase activity of Angora buck (Capra hircus ancryrensis) sperm. Ejaculates collected from three Angora bucks were used in this study. Two consecutive ejaculates from each buck were pooled and split into equal parts in four Falcon tubes. Two tubes were diluted at 37 °C and then centrifuged to remove semen plasma. After centrifugation, two sediment parts were diluted with a Tris-based extender and commercial Bioxcell extender, respectively. The remaining two parts, which were not centrifuged/washed, were diluted with the above-mentioned extenders, respectively. Diluted samples were cooled to 5 °C and frozen in 0.25-mL French straws to be stored in liquid nitrogen. Frozen straws were thawed individually at 37 °C for 20 sec in a water bath for evaluation. The semen part with centrifugation/washing in the Bioxcell extender (BC) demonstrated a higher rate of subjective motility (58.1 ± 3.0%) compared with that of groups with (TC) or without (T) centrifugation/washing in the Tris-based extender (P < 0.01). Angora buck sperm frozen with (BC) or without (B) centrifugation/washing in the Bioxcell extender demonstrated higher percentages of motility (60.6 ± 2.7% and 54.3 ± 4.8%, respectively) compared with that of groups T and TC. The postthaw progressive motility rate (22.3 ± 2.7%) was significantly greater for semen parts diluted in B compared with that of other groups. BC gave rise to a lower value of average path velocity (90.0 ± 5.2 μm/sec) compared with that of other groups (P < 0.01). For straight linear velocity and linearity index, the highest values (103.2 ± 4.7 μm/sec, 47.5 ± 1.6% and 94.8 ± 3.0 μm/sec, 44.8 ± 1.1%, respectively) were obtained from B and TC (P < 0.001). For sperm acrosome and total abnormalities, TC gave the highest values (11.2 ± 0.6% and 26.6 ± 1.5%, respectively, P < 0.01). In the group frozen in BC, the percentage of membrane integrity assessed by hypo-osmotic swelling test was higher (61.2 ± 2.2%) than that of the other groups (P < 0.001). With respect to fertility results based on 35-d pregnancy rates, BC gave a higher rate (76.5%) than that of TC (27.8%, P < 0.05). Malondialdehyde formation was found to be lower (1.64 ± 0.26 nmol/L) in BC than in the other groups after the freeze-thawing process (P < 0.001). In the semen part frozen in BC, superoxide dismutase activity was higher (0.18 ± 0.02 U/mg protein) compared with that of the other groups (P < 0.05). Further studies are required to obtain more precise results for the characterization of oxidative stress parameters and fertilizing ability in cryopreserved buck spermatozoa.     

Glucose uptake and lactate production by the autotransplanted ovary of the ewe during the luteal and follicular phases of the oestrous cycle

Two experiments were carried out on ewes with ovarian autotransplants to estimate the ovarian uptake of glucose and production of lactate. The first was carried out in the luteal phase of the oestrous cycle. Samples of carotid arterial, ovarian venous and jugular venous blood were collected simultaneously for glucose analysis. The arterial concentration of glucose (58.0 ± 5.0 mg/dL; Mean ± SEM) was significantly higher than the ovarian venous concentration (42.3 ± 2.4 mg/dL; P < 0.001). Next, a second more complete experiment was carried out in the luteal and follicular phases of the oestrous cycle. The oestrous cycle was synchronised and samples of carotid arterial, ovarian venous and jugular venous blood were collected simultaneously for glucose and lactate analysis. There were significant positive arterio-venous differences in the concentration of glucose in the luteal (5.6 ± 1.2 mg/dL, mean ± SEM; P = 0.001), early (3.1 ± 0.82 mg/d; P = 0.003) and late follicular (6.4 ± 1.3 mg/dL; P = 0.001) phases of the oestrous cycle. There was a significant negative arterio-ovarian venous difference in the concentration of lactate in only the luteal phase (-2.2 ± 0.96 mg/dL; P = 0.043).The results show significant removal of glucose from the arterial circulation during its passage through the ovary in the luteal, early follicular and late follicular phases of the oestrous cycle. Furthermore, there was lactate production in the luteal phase but not in the follicular phase suggesting that in the luteal phase of the oestrous cycle, ovarian metabolism can be anaerobic.     

Motor adaptation to prosthetic cycling in people with trans-tibial amputation

The neuromusculoskeletal system interacts with the external environment via end-segments, e.g. feet. A person with trans-tibial amputation (TTAmp) has lost a foot and ankle; hence the residuum with prosthesis becomes the new end-segment. We investigated changes in kinetics and muscle activity in TTAmps during cycling with this altered interface with the environment. Nine unilateral TTAmps and nine subjects without amputation (NoAmp) pedaled at a constant torque of 15 Nm and a constant cadence of 90 rpm (~150watts). Pedal forces and limb kinematics were used to calculate resultant joint moments. Electromyographic activity was recorded to determine its magnitude and timing. Biomechanical and EMG variables of the amputated limb were compared to those of the TTAmp sound limb and to the dominant limb in the NoAmp group using a one-way ANOVA. Results showed maximum angular displacement between the residuum and prosthesis was 4.8±1.8 deg. The amputated limb compared to sound limb and NoAmp group produced lower extensor moments averaged over the cycle about the ankle (13±2.3, 20±5.7, and 19±5.3 Nm, respectfully) and knee (8.4±5.0, 15±4.5, and 12.7±5.9 Nm, respectfully) (p<0.05). Gastrocnemius and rectus femoris peak activity in the TTAmps shifted to later in the crank cycle (by 36° and 75°, respectfully; p<0.05). These data suggest gastrocnemius was utilized as a one-joint knee flexor in combination with rectus femoris for prosthetic socket control and highlight prosthetic control as an interaction between the residuum, prosthesis and external environment.     

Defining delayed resumption of ovarian activity postpartum and its impact on subsequent reproductive performance in Holstein cows

The objectives of this study were to derive a useful case definition of delayed resumption of ovarian activity, based on factors associated with reduced fertility, and to assess its impact on subsequent reproductive performance in Holstein cows (Bos taurus). Milk samples were collected twice weekly from 219 cows from four commercial herds, and whole-milk progesterone concentrations were determined with an enzyme-linked immunosorbent assay. Ovulation was considered to have occurred 5 d before the first rise of milk progesterone concentration above the basal level. Survival analysis was used to derive a case definition of delayed resumption of ovarian activity postpartum based on factors that were predictive of reduced pregnancy rate. First postpartum ovulation occurring beyond 35 d postpartum was associated with a reduced pregnancy rate (hazard ratio [HR] = 0.50; P < 0.001) and was defined as delayed resumption of ovarian activity; overall, 75 (34.9%) cows were in this category. These cows were more likely not to conceive on first artificial insemination (odds ratio [OR] = 2.85; P = 0.01) and more likely not to become pregnant within 100 d (OR = 3.30; P = 0.001) and 210 d (OR = 3.20; P < 0.001) postpartum compared with cows with normal resumption of ovarian activity. Furthermore, 13 (6%) cows that ovulated within 35 d postpartum had a prolonged (≥14 d) interval between either first and second or second and third luteal phases postpartum. A prolonged interluteal interval was also associated with a reduced pregnancy rate (HR = 0.35; P = 0.02). Days open (mean ± SEM) were greater (P = 0.0002) in cows with delayed resumption of ovarian activity (213 ± 13 d) and in cows with prolonged interluteal interval (220 ± 37 d) than in cows with normal resumption of ovarian activity (152 ± 9 d). In conclusion, first ovulation occurring beyond 35 d postpartum was defined as delayed resumption of ovarian activity, and the first ovulation occurring within 35 d postpartum but the absence of luteal activity ≥14 d between two consecutive luteal phases was defined as a prolonged interluteal interval; both abnormalities adversely affected the subsequent reproductive performance of Holstein cows.     

Assessment of pregnancy in the late-gestation mare using digital infrared thermography

The objective of this study was to investigate use of digital infrared thermal imaging (DITI) to determine whether surface temperature gradient differences exist between pregnant and nonpregnant mares as a noncontact method to determine pregnancy status. On the day measurements were collected, each pregnant mare (n = 10; beginning at 292.4 ± 1.4 d of gestation) was paired with a nonpregnant mare (n = 17). Ambient temperature, DITI measurements (left and right flank, wither temperatures [i.e., animal surface control] and background temperature), and rectal temperatures were obtained every 7 d for 5 wk before parturition and for 3 wk after parturition. There were no differences (P > 0.10) in temperature of the left and right side within groups; therefore, data were pooled. Pregnant mares had a higher (P < 0.01) flank temperature than that of nonpregnant mares (36.0 ± 0.2 °C vs. 34.2 ± 0.2 °C, respectively). Moreover, the difference (2.4 °C) in flank temperatures between the pregnant and nonpregnant mares was greater when the ambient temperature was <19 °C. Flank and wither temperatures were positively correlated (R = 0.72; P < 0.01) and were positively correlated with ambient temperature (R = 0.48 and 0.64, respectively; P < 0.01). However, wither temperatures (skin control site) did not differ (P > 0.10) between pregnant and nonpregnant mares. In conclusion, late-gestation mares had higher flank temperatures than those of nonpregnant mares, regardless of environmental conditions, however discriminating abilities were greater when ambient temperature was lower. We inferred that DITI may have value in confirming mid- to late-gestation pregnancies in some species by noncontact means, as observed in the mare.