Spermatic and oxidative profile of domestic cat (Felis catus) epididymal sperm subjected to different cooling times (24, 48 and 72 hours)

Cooling stored epididymal samples for several days allows facilities to transport and process genetic material post‐mortem. Improvements to this practice allow the preservation of sperm from domestic cats, which are the ideal study model for wild felids. However, the modifications in spermatic features and the oxidative profile are not fully understood in cats. This information is necessary for the development of biotechniques, such as new extenders for cryopreservation. Therefore, the purpose of this study was to evaluate the spermatic and oxidative profile in samples from the epididymal cauda of domestic cats cooled at 5°C for 24, 48 and 72 hr. Spermatozoa were collected from the epididymis cauda. Evaluations consisted of computer‐assisted sperm analysis (CASA), plasma membrane integrity (eosin/nigrosin), acrosome integrity (fast green/rose bengal), sperm morphology, sperm DNA integrity (toluidine blue), mitochondrial activity (3′3 diaminobenzidine), activity of the antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD), measurement of lipid peroxidation (TBARS) and protein oxidation. A decrease in sperm motility parameters was observed after 72 hr of cooling (i.e. total and progressive) with a higher percentage of minor (37.7 ± 6.3%) and total defects (53.4 ± 6.3%). Additionally, a decrease in high mitochondrial activity (Class I: 16.6 ± 2.2%) occurred after 72 hr. The decrease in motility rates after a long cooling time probably was caused by the increase in sperm abnormalities. A long cooling time causes cold shock and mitochondrial exhaustion, but there was no observed change with the oxidative stress condition. Therefore, cat epididymal sperm stored at 5°C appear to maintain a high quality for up to 48 hr of cooling time.     

A fast,low‐cost and efficient method for the diagnosis of sperm DNA fragmentation in several species

Sperm DNA fragmentation is a condition that interferes directly in the reproductive efficiency. Currently, there are several methods for assessing the sperm DNA integrity, such as Alkaline Comet, TUNEL and Sperm Chromatin Structure Assay. However, many of these techniques are laborious and require high‐precision equipment. Thus, the development of new techniques can optimize the evaluation of sperm DNA damage. Therefore, the aim of this study was to standardize the toluidine blue (TB) stain technique for the analysis of DNA fragmentation of dog, cat, bull, stallion and ram spermatozoa. For this purpose, we used six animals of each specie (n = 30), in reproductive age. Sperm was collected by different methods according to the particularities of each species, and such samples were divided into two aliquots: a sperm sample was kept at 5°C (considered as intact sperm DNA), and the remaining samples were submitted to the induction of DNA fragmentation by exposure to ultraviolet light for 4 hr. Samples were then mixed with the intact sample to obtain known and progressive proportions of sperm with fragmented DNA (0%, 25%, 50%, 75% and 100%). Semen smears were performed and subjected to staining with TB. Blue‐stained spermatozoa were considered to have DNA fragmentation. We observed high linear regression coefficients between the expected proportion of damaged DNA and the results of TB for dog, cat, ram, bull and stallion samples. In conclusion, TB stain was considered a fast and effective technique for the study of spermatozoa DNA in several species.     

A matched observational study of survival in cats with enucleation due to diffuse iris melanoma

Although a small number of cases of feline diffuse iris melanoma have been documented to metastasize, the prognosis is not known. In this matched observational study, the survival time of 34 cats with enucleation due to histologically confirmed diffuse iris melanoma was recorded. These results are compared to the survival times of 83 age-matched control cats. Affected cats had enucleation between 2 and 10 years prior to the study. One group of control cats with eye disease had enucleation for either lymphoplasmacytic uveitis (27 cases), ocular trauma (seven cases), or endophthalmitis (four cases). In these control cats, enucleations were performed between 2 and 10 years prior to this study. Forty-five additional control cats presented for vaccination between 2 and 10 years prior to the study. The extent of diffuse iris melanoma at the time of enucleation in affected cats was graded according to the extent of involvement of ocular tissues and the invasiveness of the tumor. Affected cats have a significantly decreased survival compared with control cats and cats with extensive tumors at the time of enucleation have the lowest survival rates. Cats with tumors confined to the iris survive at the same rate as controls. These results suggests that early enucleation is important to avoid premature death, presumed to be due to cancer metastasis in cats with diffuse iris melanoma.     

Agreement of high definition oscillometry with direct arterial blood pressure measurement at different blood pressure ranges in horses under general anaesthesia

ObjectiveTo determine the agreement of high definition oscillometry (HDO) with direct arterial blood pressure measurements in normotensive, hypotensive and hypertensive horses during general anaesthesia.Study designExperimental study.AnimalsSeven healthy warmblood horses, aged 3–11 years, weighing 470–565 kg.MethodsMeasurements from a HDO device with the cuff placed around the base of the tail were compared with pressures measured invasively from the facial artery. High blood pressures were induced by intravenous (IV) administration of dobutamine (5 μg kg⁻¹ minute⁻¹) over ten minutes followed by norepinephrine (0.1 mg kg⁻¹ IV) and low pressures by increasing the inspired fraction of isoflurane and administration of nitroglycerine (0.05 mg kg⁻¹ IV). For analysis three pressure levels were determined: high (MAP>110 mmHg), normal (60 mmHgResultsA total of 245 paired measurements of systolic (SAP), mean (MAP) and diastolic (DAP) pressures were obtained. The HDO device underestimated blood pressure at hypertensive and normotensive levels and overestimated blood pressure at hypotensive levels. Best agreement was obtained for SAP and MAP within normotensive limits. At normotension, bias ± standard deviation for SAP, MAP and DAP were 0.1 ± 19.4 mmHg, 0.5 ± 14.0, 4.7 ± 15.6, respectively. At high pressure levels bias and SD were 26.1 ± 37.3 (SAP), 4.2 ± 19.4 (MAP), 1.5 ± 16.8 (DAP) and at low pressures -20.0 ± 20.9 (SAP), -11.4 ± 19.6 (MAP), -4.7 ± 20.1 (DAP), with HDO measurements at a MAP <50 mmHg often failing.Conclusion and clinical relevanceGood agreement with invasive arterial blood pressures was obtained with HDO at normotensive levels in horses. At high and low pressure ranges HDO was unreliable. Therefore, if haemodynamic instability is expected, invasive measurement remains preferable.     

The effect of footrot on body weight and wool growth of sheep

Body weight and traits associated with production of wool were measured over a 2-year period between 1985 and 1987 in south-western New South Wales in a flock of Merino wethers experimentally infected with footrot. The disease was allowed to spread freely amongst 150 of the flock but kept at very low prevalence in the remaining 50 by preventive footbathing during transmission periods. Severe, underrunning footrot had a significant adverse effect on body weight, for each year of the trial. Body weight was most severely reduced at times of the year when footrot was spreading among animals and lesions were severe. The mean body weight of the infected group at the end of the 2 years of observation was 7.3 kg (11.6%) below that of the control group. Footrot also depressed wool growth, with the mean clean fleece weight of the infected group being 0.4 kg (8%) lighter than that of the controls at each of the 2 annual shearings. There were no consistent differences between the groups for the other wool characteristics measured.     

Use of Triticum tauschii to improve yield of wheat in low-yielding environments

Triticum tauschii (Coss.) Schmal. is an ancestor of bread wheat (T. aestivum). This species has been widely used as a source ofsimply-inherited traits, but there are few reports of yield increases due tointrogression of genes from this species. Selections from F₂-derivedlines of backcross derivatives of synthetic hexaploid wheats (T.turgidum / T. tauschii) were evaluated for grain yield in diverseenvironments in southern Australia. Re-selections were made in theF₆ generation and evaluated for grain yield, yield componentsincluding grain weight, and grain growth characters in diverse environmentsin southern Australia and north-western Mexico. Re-selection was effectivein identifying lines which were higher yielding than the recurrent parent,except in full-irrigation environments. Grain yields of the selectedderivatives were highest relative to the recurrent parent in thelowest-yielding environments, which experienced terminal moisture deficitand heat stress during grain filling. The yield advantage of the derivativesin these environments was not due to a change in anthesis date orgrain-filling duration, but was manifest as increased rates of grain-filling andlarger grains, indicating that T. tauschii has outstanding potential forimproving wheat for low-yielding, drought-stressed environments.     

Comparative evaluation of the liver in dogs with a splenic mass by using ultrasonography and contrast-enhanced computed tomography

Evaluation of dogs with splenic masses to better educate owners as to the extent of the disease is a goal of many research studies. We compared the use of ultrasonography (US) and contrast-enhanced computed tomography (CT) to evaluate the accuracy of detecting hepatic neoplasia in dogs with splenic masses, independently, in series, or in parallel. No significant difference was found between US and CT. If the presence or absence of ascites, as detected with US, was used as a pretest probability of disease in our population, the positive predictive value increased to 94% if the tests were run in series, and the negative predictive value increased to 95% if the tests were run in parallel. The study showed that CT combined with US could be a valuable tool in evaluation of dogs with splenic masses.