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.     

禽流感病毒抗原超滤浓缩重要参数的动态变化

采用世界先进的切向流超滤浓缩技术，对禽流感病毒（H9N2亚型）超滤浓缩成倍体积，测定血凝价HA和半数感染量EID50。结果HA成倍增加，废液HA呈阴性，病毒回收率达100％；受宿主鸡胚系统的影响，EID50与病毒量之间不是简单的直线关系，但病毒的活性不受影响。结果表明，所用超滤系统适用于禽流感病毒抗原的浓缩。     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

Possible parachordoma in a dog

Abstract

CASE HISTORY: A 15-mm diameter mass developed in the S/C tissue overlying the right lateral gluteal muscle of a 6½-year-old female Flat-coated Retriever.

PATHOLOGICAL FINDINGS: Cytological preparations following aspiration of the mass were highly cellular and consisted of a population of large polygonal cells containing single to multiple nuclei, large prominent nucleoli, and intracytoplasmic vacuoles. Histologically, the neoplasm consisted of similar large cells surrounded by thick fibrous connective tissue trabeculae. The large polygonal cells reacted positively with antibodies against vimentin, low- and high-molecular-weight variants of cytokeratin (AE1/AE3), but not with antibodies to desmin or glial fibrillary acidic protein (GFAP).

DIAGNOSIS: The clinical, gross, histological and immunohistochemical findings are similar to those reported for parachordomas in humans. Neither recurrence nor metastases were noted 18 months after surgical excision of the mass.

CLINICAL RELEVANCE: This is the first reported case of a possible parachordoma in a dog, a benign tumour with cytological features of malignancy.     

Pharmacological differentiation of allergic and classically conditioned asthma in the guinea pig

A whole-body plethysmographic technique was developed and then used to detect experimentally induced asthma in guinea pigs and to assess pharmacological treatments of allergic and classically conditioned attacks. Inhalation of a beta adrenergic compound (isoproterenol) controlled both forms of attack. Atropine and methscopolamine, parasympathetic blocking agents, prevented conditional but not allergic attacks; diphenhydramine, an antihistamine, prevented allergic attacks; and methysergide, which blocks serotonin (which is believed to trigger human asthma), prevented neither. The guinea pig's allergic reaction is probably the result of a bronchospasm induced by histamine released in tissue of the airway by a local combination of allergen and antibody. The conditional attack is believed to be a constriction of the airway mediated by parasympathetic fibers of central origin.     

The development of multiple cutaneous inverted papilloma following ovariohysterectomy in a dog

Abstract

CASE HISTORY: Ovariohysterectomy was performed on an adult Cavalier King Charles Spaniel. The skin that had been clipped for surgery was noticed to be erythematous 8 days later.

CLINICAL AND PATHOLOGICAL FINDINGS: Poorly defined patches containing multiple papules were visible bilaterally within the clipped skin. These became larger over the following 2 weeks, and samples were collected for histology. Seven days later, the lesions were multiple raised masses, up to 5 cm in diameter. Histology revealed numerous cup-shaped epidermal proliferations extending into the dermis. The presence of keratinocytes with increased quantities of blue-grey cytoplasm, and koilocytosis suggested papillomavirul infection; Canis familiaris papillomavirus (CfPV-2) DNA was amplified from two separate samples. Complete regression was observed 8 weeks after the lesions had been initially observed.

DIAGNOSIS: Multiple inverted papilloma confined to skin that had been clipped for surgery.

CLINICAL RELEVANCE: This is the first time that the development of canine cutaneous papillomas has been associated with surgery. The nature of the association between surgery and development of the papillomas is uncertain. However, it is possible that damage to superficial skin could promote the formation of papillomas. This is the first identification of CfPV-2 in New Zealand.     

Unilateral Seminoma in a Dromedary Camel

A 10‐year‐old, clinically healthy, male dromedary camel had presented a history of progressive unilateral testicular enlargement over the past 5 years. The animal had mated with 32 females during that period; all had conceived. The sex ratio of his offspring was one male to 31 females. Ultrasound examination of the right testicle revealed a diffusely heterogeneous parenchyma with no identifiable normal testicular tissue. The enlarged testicle was surgically removed. Macroscopically, the testicle had a glistening pink surface and contained multiple soft, bulging nodules. Histopathologically, a well‐differentiated, diffuse seminoma was diagnosed. In conclusion, this study describes the fertility, sex ratio, clinical findings and ultrasonographic imaging in a male dromedary camel affected with unilateral testicular seminoma.     

The recent history of Puccinia striiformis f.sp. tritici in Denmark as revealed by disease incidence and AFLP markers

Disease observations and amplified fragment length polymorphism (AFLP) markers were used to study recent developments in the Puccinia striiformis f.sp. tritici population in Denmark. The fungus appeared spontaneously at 10 locations in Denmark in 1997 after it was not observed under natural conditions in 1996. The pattern of disease development and prevailing winds suggested that the fungus reappeared by airborne spores from the south or west. In 1998, disease incidence was more evenly distributed throughout the country. Forty-eight single lesion isolates were collected from most crops where the disease was observed in these years; all except one from 1997 belonged to two pathotypes that were not previously detected in the country, and both possessed the newly discovered Yr17 virulence. The isolates were characterized with AFLP markers together with 28 isolates representing eight of 13 pathotypes observed prior to 1996. Initial screening of 240 Pst I/ Mse I AFLP primer combinations on four isolates showed that a primer combination, on average, revealed 0·4 polymorphisms between any isolate pair. A selection of 21 primer combinations resulted in 28 AFLP markers, which revealed 16 AFLP phenotypes among all 76 isolates. The two Yr17- virulent pathotypes consisted of three AFLP phenotypes, which were observed in both 1997 and 1998; the two most frequent AFLP phenotypes occurred at most sampling locations and often within the same crop. AFLP diversity was larger among samples collected prior to 1996, and also in this period most AFLP phenotypes were observed at different sampling locations. These results are consistent with the features of an entirely asexually reproducing pathogen dispersed by aerial spores across large areas.     

Studies of yield losses I. The self-limiting effect of injurious or competitive organisms on crop-yield

Summary Four hypothetical models illustrate how competition between pathogens diminishes the effect of individuals as numbers increase. As a result the relation between crop yield or growth and the initial population level is not linear. The effect on yield falls off as the initial population increases or when graphed the yield curve tends to become horizontal.Samenvatting Aan vier modellen wordt duidelijk gemaakt, hoe concurrentie tussen schadelijke organismen de invloed van het individu op de waardplant kan verminderen. Als gevolg daarvan zal de relatie tussen het aantal organismen en de oogstderving of de groei niet rechtlijning zijn. Het effect wordt relatief minder als het aantal schadelijke organismen stijgt. In een grafiek heeft de lijn de neiging om horizontaal te worden bij hogere aantallen organismen, bijvoorbeeld fig. 6 (oogst verticaal, schadelijke organismen horizontaal).For the mathematics the reader is referred to the appendix.