Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Immunohistochemical Expression of Growth Factors in the Follicular Wall of Normal and Cystic Ovaries of Sows

The expression of growth factors was evaluated immunohistochemically in normal and cystic ovaries of sows. The immunohistochemically stained area (IHCSA) was quantified by image analysis to analyse the expression of these proteins in the follicular wall of secondary, tertiary and cystic follicles. IGF‐I immunoreactivity was strong in the granulosa cell layer (GC), moderate in the theca interna (TI) and mild in the theca externa (TE) of the normal follicles. There was severe reduction of the labelling to IGF‐I in the GC of the follicular and luteinized cysts. In the normal follicles, the reactivity for IGF‐II was very similar to pattern noted in IGF‐I. There was reduction of the IHCSAs in the GC of the follicular and luteinized cysts, but the decrease was not significant. The staining of the IGF‐II in the TI and TE of the cysts was increased, in comparison with normal follicles. The IHCSAs for VEGF were higher in the GC and TE of the normal follicles in contrast to TI, but this difference was noted only in the tertiary follicle. The VEGF reactivity increased in the GC of the cysts, in relation to normal follicles. The results of the current study show that the formation of ovarian cysts in sows is associated with alterations in the immunohistochemical expression of some growth factors.     

The Influence of Oxygen Tension on Cumulus Cell Viability of Canine COCs Matured in High-Glucose Medium

High in vitro oxygen (O₂) tensions are associated with enhanced levels of reactive oxygen species and cumulus oocyte complex (COC) apoptosis. The objective of this study was to determine the influence of O₂ tension on cumulus cell (CC) viability from canine oocytes. Cumulus oocyte complexes were distributed into three groups (CG, T20 and T5) and two O₂ tension levels (20% and 5%). The control group (CG) was matured in vitro in a humidified atmosphere with 5% CO₂ in air in TCM199 with 26.19 m m sodium bicarbonate, 10% (v/v) foetal calf serum (FCS), 0.10 m m gentamicin, 0.20 m m pyruvic acid, 20 μg/ml oestradiol, 0.5 μg/ml follicle-stimulating hormone, 0.03 IU/ml human chorionic gonadotropin, and 1.0 μg/ml human somatotropin. Groups T20 and T5 were matured under 20% or 5% O₂ tensions respectively in a high-glucose medium, without FCS. T20 and T5 were as CG, and supplemented with 0.1% Polyvinyl Alcohol, and 5.5 m m glucose. After 48 h of IVM, CCs from COCs were stained with propidium iodide (1.50 m m ). The results showed that viability of CCs (cytoplasmic features and nuclear morphological integrity) was different for the three groups. Rates of apoptosis were at 57.9% (521/900) for CG, 54.4% (490/900) for T20 and 38.9% (350/900) for T5 (p < 0.001). Predominant features in apoptotic cells (n = 1361) were DNA nuclear fragments (94.0%). It was concluded that CCs of canine COCs cultured in high-glucose medium showed significantly less apoptosis than those cultured in medium with FCS. Low O₂ tension was efficient in reducing apoptosis in canine CCs.     

Use of radiographic measures and three-dimensional computed tomographic imaging in surgical correction of an antebrachial deformity in a dog

CASE DESCRIPTION: A 1-year-old 7.4-kg (16.3-lb) castrated male mixed-breed dog was evaluated because of intermittent lameness and an antebrachial angular limb deformity. CLINICAL FINDINGS: The left forelimb had gross antebrachial external rotation (approx 90 degrees ) and marked procurvatum. Radiography revealed a severe partially compensated biapical antebrachial angular limb deformity. Measurements of medial proximal radial angle (MPRA) and lateral distal radial angle (LDRA) were obtained from orthogonal radiographs of the proximal and distal segments of the radius, respectively. Elbow joint-to-carpus translation was quantified. Deformities were localized and quantified by the center of rotation of angulation (CORA) method. Computed tomographic 3-dimensional image reconstructions of the antebrachium and carpus were completed to create 3 life-size stereolithographic models. TREATMENT AND OUTCOME: 2 closing wedge radial osteotomies were performed at the level of the CORAs and stabilized with bone plates and screws. RESULTS: Frontal and sagittal plane alignments were corrected to 8 degrees and 15 degrees , respectively (reference limits, 0 degrees to 8 degrees and 8 degrees to 35 degrees , respectively). The MPRA was corrected from 55 degrees to 68 degrees , and LDRA was corrected from 32 degrees to 76 degrees (values considered normal are approx 85 degrees and 87 degrees , respectively). Elbow joint-to-carpus translation was improved by 42.5%. After 8 weeks, radiography revealed bone union. Owners considered the outcome acceptable, on the basis of limb appearance and lack of lameness at 1 year after surgery. CONCLUSIONS AND CLINICAL RELEVANCE: A segmental radiographic planning technique combined with the CORA method, computed tomography, and stereolithography may be useful in the characterization of and planning corrective surgery for forelimb deformities in dogs.     

蛋雏鸡传染性喉气管炎的诊治

鸡传染性喉气管炎(ILT)是由鸡传染性喉气管炎病毒引起的以剧烈咳嗽、气喘、高度呼吸困难和气管内有血样渗出物等为特征的一种急性呼吸道传染病.育成蛋鸡感染该病多有报道,而蛋雏鸡感染发病比较少见.本文以一起48日龄海兰褐蛋雏鸡传染性喉气管炎病例为题材,从发病情况、临床症状、病理剖检变化、诊断、防治等几个方面对该病做了详尽阐述.