Effects of Egg Yolk and Cooling Rate on the Survival of Refrigerated Red Deer (Cervus elaphus hispanicus) Epididymal Spermatozoa

Egg yolk is a common component to sperm refrigeration for most of the deer species, the role of which is to protect sperm membranes against cold shock. In addition, there have been many studies of conservation of ejaculated semen from stags, but few have been reported for epididymal spermatozoa. This work was designed to investigate the combined effects of cooling rates (slow: 0.23 degrees C/min vs rapid: 4.2 degrees C/min) from room temperature to 5 degrees C, and egg-yolk concentration (0, 5 or 20%) in the extender on the survival of Iberian red deer epididymal spermatozoa refrigerated at 5 degrees C. Heterospermic sperm samples were diluted to a final sperm concentration approximately 400x10(6) sperm/ml with a Tris-citrate-fructose (TCF)-egg-yolk diluent. Sperm quality was in vitro judged by microscopic assessments of individual sperm motility [sperm motility index (SMI)], and of plasma membrane (hypo-osmotic swelling test) and acrosome (NAR) integrities. Our results first showed that the presence of egg yolk in the extender significantly improves (p=0.01) the viability and sperm motility after sperm dilution. In addition, acrosome and plasma membrane integrities post-refrigeration did not differ significantly between cooling procedures; however, the SMI differed significantly between cooling procedures (slow: 46.6% vs rapid: 50.0%; p=0.01). Our results also showed that sperm quality was significantly (p<0.01) affected by the combined effects of egg-yolk concentration and cooling procedure, being rapid cooling with 20% of egg yolk the most suitable combination for epididymal sperm refrigeration. In conclusion, egg-yolk improved red deer epididymal spermatozoa characteristics after dilution. Rapid cooling protocol using TCF with 20% egg-yolk significantly improved sperm motility of red deer epididymal spermatozoa after cooling.     

Immunohistochemistry Diagnosis of an Ovarian Dysgerminoma in One Bitch

An ovarian enlargement (diameter, 8 cm) was identified and surgically excised from a 5-year-old female dog. Microscopic examination of the multinodular neoplasm revealed sheets of polygonal neoplastic cells with large nuclei, frequent mitosis, necrosis and haemorrhage. Immunohistochemically, the neoplastic cells were positive for vimentin and alkaline phosphatase but were negative for CD3, CD79a, cytokeratin, alpha-fetoprotein, inhibin-α and S-100. The histopathological diagnosis of the mass was unilateral ovarian dysgerminoma.     

Identification of Sperm-Head Morphometric Subpopulations in Iberian Red Deer Epididymal Sperm Samples

Computer-assisted sperm morphometry analysis (CASMA) was used in this study to identify sperm morphometric subpopulations in Iberian red deer epididymal sperm samples. Epididymal sperm samples were collected from 37 mature stags and were divided. One portion was diluted in a Tris–citrate–egg yolk medium. A microscope slide was prepared from single extended sperm samples prior to freezing. The remainder of each sample was frozen in nitrogen vapours using a conventional protocol. After thawing, sperm smears were prepared as described for extended samples. All slides were air-dried and stained with Hemacolor^®. The sperm-head dimensions for a minimum of 145 sperm-heads were analyzed from each sample by means of the Sperm-Class Analyser^®, and the mean measurements recorded. Each sperm-head was measured for four primary sperm-head parameters, and five parameters of head shape. All sperm morphometric parameters evaluated were placed in a statistical database and a multivariate cluster analysis was performed. The clustering analyses, based on 10 867 individual spermatozoa, revealed the existence of three subpopulations (SP₁, SP₂, SP₃) of spermatozoa with different morphometric characteristics (p  <  0.001). The proportion of spermatozoa present in any of the three subpopulations remained constant (p  >  0.05) through the cryopreservation process. Pre-freeze and post-thaw sperm quality was in vitro evaluated by microscopic assessments of individual sperm motility and of plasma membrane and acrosome integrities. In conclusion, our results show that applying the CASMA techniques and multivariate cluster analyses, it was possible to determine that three subtle subpopulations of spermatozoa with different morphometric characteristics coexist in red deer semen.     

Communicating the risks of handling bats: analysing approaches used by Australian stakeholders in the context of Australian bat lyssavirus

Australian bat lyssavirus (ABLV) is a member of the Lyssavirus genus of the Rhabdoviridae family and is found in Australian bat species. It is of public health concern because of the rabies-like syndrome it causes in humans, resulting in government health and wildlife agencies using varied communication approaches to inform targeted audiences about zoonotic risks associated with handling bats. Despite these warnings, the number of reports of human-bat interactions remains high. This paper details a survey conducted to analyse the approaches utilised by a range of stakeholders to educate and communicate warnings to their target audiences. The survey focused on identifying the target audiences, communication methods used, along with the message frequency, content, and perceived effectiveness. Analysis of the top three messages delivered by stakeholders revealed that over half were information-focused messages and over a third, instruction-focused. Stakeholders identified the need to balance messaging about bat handling risks with information regarding the vulnerable status of bats and their environmental significance. Whilst the most common and (perceived) effective method of communication was one-on-one discussions, it was also identified to be ineffective for targeting mass audiences leading stakeholders to recognise the need to adapt to more efficient means of communication. The outcomes of this study may be useful to improve risk communication strategies regarding ABLV in Australia.     

In vivo imaging reveals novel aspects of retinal disease progression in Rs1h(-/Y) mice but no therapeutic effect of carbonic anhydrase inhibition

Purpose X-linked juvenile retinoschisis (XLRS) is the most common juvenile maculopathy in men and is caused by mutations in the gene encoding retinoschisin (RS1). Evidence in the literature on the therapeutic effect of carboanhydrase inhibitors (CAIs) to treat schisis formation in the retina has remained equivocal. Here, we evaluate the effect of the CAI dorzolamide on the structural and functional disease progression in the mouse model for XLRS (Rs1h(-/y) ). Methods Rs1h ( -/y ) mice were treated unilaterally with dorzolamide eye drops (Trusopt(?) 20?mg/mL) every 12?h for 2?weeks starting on postnatal day 14 (n?=?27). Changes of retinal structure were monitored by confocal scanning laser ophthalmoscopy and spectral domain optical coherence tomography 12?h, 14?days, 4?weeks, 2?months, and 6?months after completion of the treatment. Results Schisis formation (peak at 3?months) preceded photoreceptor degeneration and hyper-fluorescence (peak at 7?months). Structural pathology was most severe in the superior hemi-retina with previously unreported hyper-fluorescent lesions. Quantitative analysis showed no significant differences regarding the inner or outer retinal thickness of the treated vs. untreated eyes 12?h after the completion of treatment (IRT(12?h) =?-1.29?±?1.89?μm; ORT(12?h) =?0.61?±?2.08?μm; mean?±?95%CI) or at any later time point. Conclusion Time line analysis after short-term treatment with CAI failed to show short-, intermediate-, or long-term evidence of structural improvement in Rs1h(-/y) mice. Schisis formation in the inner retina peaked at the age of 3?months and was followed by photoreceptor degeneration predominantly in the superior hemi-retina. Previously unreported hyper-fluorescent lesions co-register with structural retinal pathologies.     

Chondrodysplasia in Australian Dexter cattle

Objective To describe the occurrence of chondrodysplasia in Australian Dexter cattle.
Design A pathological and genetic case report.
Procedure Congenital lethal chondrodysplasia was studied in two female Dexter foetuses aborted mid to late gestation. Clinicopathological findings including histological changes in limb bones, and analysis of pedigree information were evaluated.
Results Characteristic features of congenital lethal chon-drodysplasia (Dexter bulldog) include abortion, disproportionate dwarfism, a short vertebral column, marked micromelia, a relatively large head with retruded muzzle, cleft palate and protruding tongue and a large abdominal hernia. Histological changes in limb bones are consistent with failure of endochondral ossification. Dexter chondrodysplasia is considered to be inherited in an incompletely dominant manner with the homozygous form producing the congenital lethal condition. A preliminary minimum estimate of heterozy-gote frequency is 19% within the registered Australian Dexter herd, based on analysis of the contribution of three obligate heterozygotes whose semen has been widely used by artificial insemination in Australia.
Conclusion Dexter chondrodysplasia is present in Australian cattle and further cases of the homozygous form, congenital lethal chondrodysplasia, are likely to occur.
Recommendation It is requested that spleen and liver tissue from bulldog foetuses and blood from their parents be collected to assist research into Dexter chondrodysplasia.     

Observation of Interference Between Two Bose Condensates

Interference between two freely expanding Bose-Einstein condensates has been observed. Two condensates separated by approximately 40 micrometers were created by evaporatively cooling sodium atoms in a double-well potential formed by magnetic and optical forces. High-contrast matter-wave interference fringes with a period of approximately 15 micrometers were observed after switching off the potential and letting the condensates expand for 40 milliseconds and overlap. This demonstrates that Bose condensed atoms are "laser-like"; that is, they are coherent and show long-range correlations. These results have direct implications for the atom laser and the Josephson effect for atoms.     

Tetravalent uranium in calcite

X-ray absorption spectroscopy and x-ray fluorescence microprobe studies of 35-million-year-old calcite from a Mississippi Valley-type zinc ore deposit indicate substitution of tetravalent uranium for divalent calcium. Thus, tetravalent uranium has a stable location in calcite deposited under reducing conditions. This result validates uranium-series dating methods (including uranium/lead dating) for ancient calcite and shows that calcite provides a sink for uranium in deep groundwater aquifers and anoxic lacustrine and marine basins.     

Effects of supplemental dietary vitamin C on quality of semen from Nile tilapia (Oreochromis niloticus) breeders

The objective of this study was to evaluate the effects of vitamin C on growth and quality of semen from Oreochromis niloticus breeders. One hundred and sixty males were fed with different levels of vitamin C (0, 261, 599 and 942 mg/kg diet). The higher weight values were recorded for 599 (166 g) and 942 (175 g) mg of vitamin C/kg diet. Sperm motility, vigour and concentration were higher with 599 and 942 mg of vitamin C/kg diet. The semen volume, gonadosomatic index and plasma protein data from the last week showed a direct relationship with increasing levels of vitamin C. No changes were observed in the hepatosomatic index and blood glucose. The haematocrit and erythrocyte showed higher values estimated by equations derived at 850 and 638 mg vitamin C/kg diet, respectively. The leucocytes were inversely proportional to the increasing levels of vitamin C. After 100 days of feeding, animals fed the diet containing 942 mg vitamin C/kg diet had higher sperm motility, linearity, curvilinear velocity, straight line velocity and average path velocity (p < .05). Higher values of beat cross‐frequency were observed in broodfish fed diets containing 942 and 599 mg vitamin C/kg. The different vitamin C levels did not cause differences in straightness, lateral head displacement and sperm morphology. For Nile tilapia males on intensive rearing and handling conditions, vitamin C levels between 599 and 942 mg/kg may be used for a better performance and quality of semen.