Identification of Apoptotic Bodies in Equine Semen     

AB Caselles  A Miro‐Moran  A Morillo Rodriguez  JM Gallardo Bolaños  C Ortega‐Ferrusola  GM Salido  FJ Peña  JA Tapia  IM Aparicio 《Reproduction in domestic animals》2014,49(2):254-262

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.  相似文献   

Colloid Centrifugation Selects Normal Spermatozoa from Polymorphic Bull Ejaculates: A Case Study     

JM Morrell  H Rodriguez‐Martinez  M Andersson 《Reproduction in domestic animals》2014,49(2):281-284

Semen from a Western Finncattle bull exhibiting a highly polymorphic spermiogram was processed by colloid centrifugation using Androcoll‐B, a species‐specific silane‐coated silica colloid. In the first experiment, Single Layer Centrifugation (SLC) was used to identify which density colloids were needed to separate different cell populations. Colloids of the two chosen densities were then used in a density gradient resulting in two sperm subpopulations, one containing nearly all normally sized spermatozoa and the other enriched for the macrocephalic spermatozoa. Microcephalic spermatozoa did not appear in either of the selected subpopulations. Using a combination of SLC and DGC with this species‐specific colloid, it was possible to separate the spermatozoa into different subpopulations, that is, a subpopulation containing nearly all normally sized spermatozoa, and another one enriched for the macrocephalic spermatozoa. Thus, colloid centrifugation could be used to select sufficient normal spermatozoa from a highly polymorphic ejaculate for AI, if desired.  相似文献   

Anti‐Mullerian Hormone Concentration and Antral Ovarian Follicle Population in Murrah Heifers Compared to Holstein and Gyr Kept Under the Same Management     

JM Baldrighi  MF Sá Filho  EOS Batista  RNVR Lopes  JA Visintin  PS Baruselli  MEOA Assumpção 《Reproduction in domestic animals》2014,49(6):1015-1020

This study was performed to evaluate plasma concentrations of anti‐Mullerian hormone (AMH) and the ovarian antral follicle population (AFP) in different genetic groups. Cyclic heifers (13 Bubalus bubalis [Murrah]; 15 Bos taurus [Holstein] and 10 Bos indicus [Gyr]) were maintained under the same management and were synchronized with two doses of 150 μg IM d‐cloprostenol administered 14 days apart. After the second d‐cloprostenol treatment, heifers had their ovaries scanned daily by ultrasound to define the day of ovulation. On the same day, the AFP was determined and a plasma sample was collected to measure AMH. Murrah heifers had less AFP (25.6 ± 2.1 follicles; p = 0.01) and plasma AMH concentration (0.18 ± 0.03 ng/ml; p < 0.001) than Gyr (60.0 ± 12.2 follicles and 0.60 ± 0.12 ng/ml of AMH); however, data were similar when compared to Holstein (35.9 ± 6.8 follicles and 0.24 ± 0.06 ng/ml of AMH) heifers. Regardless of genetic background, there was a positive relationship between the AFP and plasmatic AMH concentration (Murrah [r = 0.62; p < 0.01], Holstein [r = 0.66; p < 0.001] and Gyr [r = 0.88; p < 0.001]). Also, when heifers were classified according to high‐ or low‐AMH concentration based on the average within each genetic group, high‐AMH heifers had greater (p < 0.0001) AFP than low‐AMH heifers. In conclusion, both Murrah and Holstein heifers presented lower plasma AMH concentration and AFP when compared to Gyr.  相似文献   

Effect of Overnight Staining on the Quality of Flow Cytometric Sorted Stallion Sperm: Comparison with Tradtitional Protocols     

CM Balao da Silva  C Ortega Ferrusola  JM Gallardo Bolaños  M Plaza Dávila  P Martín‐Muñoz  JM Morrell  H Rodriguez Martínez  FJ Peña 《Reproduction in domestic animals》2014,49(6):1021-1027

Flow cytometry is considered the only reliable method for the separation of X and Y chromosome bearing spermatozoa in equines. The MoFlo SX DP sorter is highly efficient, allowing the production of foals of the desired sex. However, to achieve acceptable pregnancy rates the currently used protocol requires working with fresh semen obtained close to, or at, the sorting facility. An alternative protocol was tested during two consecutive breeding seasons. Fresh stallion semen was cooled for 20 h, during which staining with Hoechst 33342 took place. On the following day, this sample was flow sorted and compared with spermatozoa from the same ejaculate that had been sexed on the previous day. All sperm parameters evaluated remained unchanged when fresh sorted and refrigerated sorted semen were compared. Pre‐sorting storage at 5°C did not alter sperm velocities nor kinetics, viability or membrane permeability, production of reactive oxygen species, mitochondrial membrane potential or DNA fragmentation index of the sorted sample. The findings open for the possibility of using semen from stallions housed far from the sorting facilities. Processed and stained sperm could be shipped refrigerated on the previous day, sorted and inseminated on the next day.  相似文献   

Standing Laparoscopic Peritoneal Flap Hernioplasty of the Vaginal Rings does not Modify the Sperm Production and Motility Characteristics in Intact Male Horses     

LA Gracia‐Calvo  LJ Ezquerra  M Martín‐Cuervo  ME Durán  H Tapio  JM Gallardo  FJ Peña  C Ortega‐Ferrusola 《Reproduction in domestic animals》2014,49(6):1043-1048

Laparoscopic hernioplasty techniques have been developed in the recent years to avoid the recurrence of inguinal hernias and to spare the testicles for breeding purposes in stallions. However, there have been no previous comprehensive and systematic studies of the reproductive outcomes and prognoses for stallions after inguinal hernioplasty. Therefore, the objective of this study was to assess the possible effects of one of these techniques (standing laparoscopic peritoneal flap hernioplasty) on the sperm production and motility characteristics of six healthy stallions that received this procedure based on 1‐year follow‐ups. There were no significant differences in the measured sperm variables (assessments based on the DSO, MOT, PMOT, VSL, VCL and VAP) during 1‐year follow‐ups.  相似文献   

Changes in Bull Sperm Kinematics after Single Layer Centrifugation     

Y Yulnawati  MC Abraham  D Laskowski  A Johannisson  JM Morrell 《Reproduction in domestic animals》2014,49(6):954-956

The objective of this study was to evaluate bull sperm kinematics after centrifugation through a single layer of a colloid [Single Layer Centrifugation (SLC)]. Ejaculates from 20 bulls were extended and stored at 4–6°C for 24 h during transport to the laboratory for SLC through Androcoll‐B, followed by measurement of sperm kinematics in all samples. Total motility (86% and 88% for uncentrifuged and SLC samples, respectively) and progressive motility (84% for both the groups) were similar (p > 0.05). In contrast, straightness (STR) (0.65 vs 0.69), linearity (LIN) (0.32 vs 0.35) and beat cross frequency (BCF) (22.3 vs 23.6 Hz) were significantly higher in the SLC‐selected samples than in the uncentrifuged samples, whereas velocity of the average path (VAP) (95 vs 90 μm/s), curvilinear velocity (VCL) (192 vs 180 μm/s), amplitude of lateral head deviation (ALH) (7 μm vs 6.5 μm) and hypermotility (49% vs 38%) were significantly decreased. The kinematics of the samples with the poorest motility was improved most by SLC. In conclusion, even though SLC had no direct effect on total and progressive motility, it appeared to have a positive influence on several other kinematic parameters that may be important for fertilization after artificial insemination.  相似文献   

Response of cotton water stress indicators to soil salinity     

T. A. Howell  J. L. Hatfield  J. D. Rhoades  M. Meron 《Irrigation Science》1984,5(1):25-36

Summary A field study was conducted on cotton (Gossypium hirsutum L. c.v. Acala SJ-2) to investigate the effects of soil salinity on the responses of stress indices derived from canopy temperature, leaf diffusion resistance and leaf water potential. The four salinity treatments used in this study were obtained by mixtures of aqueduct and well water to provide mean soil water electrical conductivities of 17, 27, 32 and 38 dS/m in the upper 0.6 m of soil profile. The study was conducted on a sandy loam saline-alkali soil in the lower San Joaquin Valley of California on 30 July 1981, when the soil profile was adequately irrigated to remove any interference of soil matric potential on the stress measurements. Measurements of canopy temperature, leaf water potential and leaf diffusion resistance were made hourly throughout the day.Crop water stress index (CWSI) estimates derived from canopy temperature measurements in the least saline treatment had values similar to those found for cotton grown under minimum salinity profiles. Throughout the course of the day the treatments affected CWSI values with the maximum differences occurring in mid-afternoon. Salinity induced differences were also evident in the leaf diffusion resistance and leaf water potential measurements. Vapor pressure deficit was found to indicate the evaporative demand at which cotton could maintain potential water use for the various soil salinity levels studied. At vapor pressure deficits greater than 5 kPa, cotton would appear stressed at in situ soil water electrical conductivities exceeding 15 dS/m. The CWSI was as sensitive to osmotic stress as other, more traditional plant measures, provided a broader spatial resolution and appeared to be a practical tool for assessing osmotic stress occurring within irrigated cotton fields.  相似文献   

Apoplastic pH and monolignol addition rate effects on lignin formation and cell wall degradability in maize     

Grabber JH  Hatfield RD  Ralph J 《Journal of agricultural and food chemistry》2003,51(17):4984-4989

Monolignol polymerization rate and apoplastic pH and may influence the formation of lignin and its interactions in cell walls. Primary maize walls were artificially lignified by gradual "end-wise" or rapid "bulk" polymerization of coniferyl alcohol at pH 4 or 5.5. Lignification efficiency was greatest for end-wise polymers at pH 5.5 (90-98%), intermediate for bulk polymers formed at either pH (54-82%), and lowest for end-wise polymers at pH 4 (41-53%). End-wise polymers had about 2.2-fold more ether inter-unit linkages and 70% fewer end-groups than bulk polymers. Low pH enhanced the formation of ether linkages in end-wise but not in bulk polymers. Differences in lignin structure did not influence the enzymatic degradability of cell walls, but lowering apoplastic pH from 5.5 to 4.0 during lignification reduced cell wall degradability by 25%. Further studies indicated this pH-dependent depression in degradability was related to cell wall cross-links formed via lignin quinone methide intermediates.  相似文献   

Effects of panicles on infrared thermometer measurements of canopy temperature in wheat     

J.L Hatfield  P.J Pinter  E Chasseray  C.E Ezra  R.J Reginato  S.B Idso  R.D Jackson 《Agricultural and Forest Meteorology》1984,32(2):97-105

A study was conducted in Phoenix, AZ on stressed and unstressed field plots of Anza wheat (Triticum aestivum L.) on an Avondale loam soil (a fine, loamy, mixed calcareous hyperthermic Anthropic Torrifluvent) to determine effects of panicles on the apparent canopy temperature and their consequent impact on the estimation of crop stress. The panicles were removed from a 1.5 × 4-m sample of each plot by extracting the peduncle from the upper sheath. For each treatment canopy radiative temperature measurements were made from vertical and oblique angles (30° from the horizontal), using an 8° field-of-view (FOV) infrared thermometer, at half-hour intervals from sunrise to sunset on 20, 22, and 30 April. Complementary measurements included leaf water potential and leaf diffusive resistance.Apparent canopy temperatures obtained from the oblique view of the canopy with panicles and under well-watered conditions were 2°C warmer than those of the unstressed canopy without panicles. In the stressed plot the canopy with panicles was 1°C cooler than that without panicles, but this effect was only noticed around 1200 MST. The temperature difference between viewing angles was apparently caused by different percentages of panicle area viewed by the radiometer. In the vertical view panicles contributed to 3% of the total viewed area while at the 30° oblique view panicles comprised 40% of the area. Since energy balance calculations of a non-transpiring cylinder with dimensions similar to a typical wheat panicle showed its temperature would remain very close to that of the surrounding air, canopy temperatures were adjusted for the proportion of panicles viewed assuming they were in equilibrium with air temperature. Results showed the corrected canopy temperatures of the canopy with panicles were the same as those measured in the canopy without panicles. Such a correction is necessary to avoid an overestimate of the stress level and an underestimate of differences between treatments. Crops with non-transpiring and/or well-ventilated morphological structures above the foliage will require this correction if radiative canopy temperatures are to be used in irrigation management programs or stress detection studies.  相似文献   

Model studies of ferulate-coniferyl alcohol cross-product formation in primary maize walls: implications for lignification in grasses     

Grabber JH  Ralph J  Hatfield RD 《Journal of agricultural and food chemistry》2002,50(21):6008-6016

Ferulate and diferulates mediate cell wall cross-linking in grasses, but little is known about their cross-coupling reactions with monolignols and their role in lignin formation in primary cell walls. Feruloylated primary walls of maize were artificially lignified and then saponified to release ferulate and diferulates and their cross-products with coniferyl alcohol for analysis by GC-FID, GC-MS, and NMR spectroscopy. Ferulate and 5-5-coupled diferulate had a greater propensity than 8-coupled diferulates to copolymerize with coniferyl alcohol, forming mostly 4-O-beta' and 8-beta' and some 8-O-4' and 8-5' cross-coupled structures. Some 8-beta' structures de-esterified from xylans, but these cross-links were subsequently replaced as 8-coupled diferulates formed stable cross-coupled structures with lignin. Based on the incorporation kinetics of ferulate and diferulates and the predicted growth of lignin, cross-products formed at the onset of lignification acted as nucleation sites for lignin polymerization.  相似文献