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.     

Lipophilic hydroxytyrosyl esters. Antioxidant activity in lipid matrices and biological systems

Antioxidant activities of lipophilic hydroxytyrosyl acetate, palmitate, oleate, and linoleate were compared with those of hydroxytyrosol, alpha-tocopherol, and butylhydroxytoluene (BHT) in both glyceridic matrix and biological systems. Aliquots of a glyceridic matrix spiked with various concentrations of antioxidant were subjected to accelerated oxidation in a Rancimat apparatus operated at 90 degrees C. The relationships between induction time (IT) and antioxidant concentration (mmol/kg) presented by hydroxytyrosol and hydroxytyrosyl acetate, palmitate, oleate, and linoleate were similar. Hydroxytyrosol and its esters showed greater antioxidant activity than alpha-tocopherol or BHT. We also evaluated the capacity of hydroxytyrosyl esters to protect proteins and lipids against oxidation caused by peroxyl radicals, using a brain homogenate as an ex vivo model. All tested compounds showed a protective effect in these systems, which was greater in preventing the generation of carbonyl groups in protein than of malondialdehyde in lipid. Inclusion of a lipophilic chain in the hydroxytyrosol molecule enhanced its antioxidant capacities in this biological model.     

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.     

猪败血性链球菌与附红细胞体混合感染病例

猪败血性链球菌病是猪链球菌感染引起的一种急性败血性传染病,猪附红细胞体病是由猪附红细胞体寄生于猪红细胞而引起的一种散在的热性、溶血性传染病.本文以某猪场这2种病混合感染病例为题材,从发病基本情况、临床症状、剖检变化、实验室检验、防治措施等6个方面对该混合感染病例做了详尽的阐述.     

Failure of passive transfer in foals: incidence and outcome on four studs in New South Wales

Objective To determine the regional incidence and effectiveness of treatment of failure of passive transfer (FPT) in foals. Design A study of disease incidence. Animals Eighty-eight foals and 57 mares from four studs in the practice area of the Rural Veterinary Centre were tested. Procedure Foals were tested for their serum IgG and total serum protein (TSP) concentration within the first 72 hours of life. Colostrum was collected from mares and specific gravity determined. FPT and partial failure of passive transfer (PFPT) of immunoglobulins was diagnosed when serum IgG concentrations were < 4 g/L and 4 to 8 g/L respectively. Owners of foals diagnosed with FPT were offered treatment with 1 to 2 L plasma (TSP > 70 g/L); 9 (64%) of the affected foals were treated. Results Fourteen foals (16%) had FPT whereas 15 (17%) had PFPT. There were significant differences between the mean TSP concentration in foals with FPT (42.6 ± 4.2 g/L), PFPT (48.1 ± 3.9 g/L) and those acquiring adequate passive immunity (58.9 ± 5.5 g/L) (P < 0.01). Sixteen (29%) mares had pre-suck colostral specific gravity < 1.060 and 12 (71%) foals raised by these mares had FPT or PFPT. The incidence of severe disease (categorised by a sepsis score > 11, positive culture of bacteria from blood or disease requiring hospitalisation) in all foals in the first 2 months of life was 10%. However, none of the nine foals with FPT that received plasma experienced severe disease. In contrast, foals with PFPT had an increased susceptibility to severe disease (P < 0.001) when compared with normal foals. Conclusion Treatment of foals with FPT may reduce the subsequent incidence of severe disease. Pre-suck colostral specific gravity and foal TSP may be used to predict the likelihood of FPT and PFPT. Even though the number of foals studied is small the results highlight the importance of optimal management practices in reducing the incidence of FPT and disease associated with this process.     

Tracking the long-term decline and recovery of an isolated population

Effects of small population size and reduced genetic variation on the viability of wild animal populations remain controversial. During a 35-year study of a remnant population of greater prairie chickens, population size decreased from 2000 individuals in 1962 to fewer than 50 by 1994. Concurrently, both fitness, as measured by fertility and hatching rates of eggs, and genetic diversity declined significantly. Conservation measures initiated in 1992 with translocations of birds from large, genetically diverse populations restored egg viability. Thus, sufficient genetic resources appear to be critical for maintaining populations of greater prairie chickens.     

Intraperitoneal Insemination in Mammals: A Review

This review focuses on factors associated with the development of intraperitoneal insemination in mammals. Findings to date indicate that fertility improves as the sperm cell concentration rises, but that the optimal sperm number differs in each species. Sperm washing before intraperitoneal insemination favours fertility. Peritoneal fluid shows a variable effect on spermatozoa, depending on the hormonal status of the female. The optimal time for insemination appears to be just prior to ovulation. The technique may be performed either through the abdominal or the vaginal wall. Verification of sperm deposition in the proximity of the ovaries improves fertility rates. Although associated with some risk of infection and an immune reaction against spermatozoa, the intraperitoneal technique rarely gives rise to severe anaphylactic shock, peritonitis, adhesion formation and the production of anti‐sperm antibodies and these complications may be prevented by adequate sperm pretreatment and antibiotic therapy. The success of intraperitoneal insemination in humans, with results comparable with those of intrauterine insemination in the treatment of infertility, suggest the potential use of this technique in domestic mammals, especially in those in which intrauterine insemination poses practical difficulties. Some of the methods applied in human intraperitoneal insemination, such as confirming the position of the needle in the peritoneal cavity, and sperm pre‐treatments might also improve results in domestic species. Conversely, the use of the animal model should help to develop some aspects of this technique in humans.