Flow cytometric analysis of T cell response in mice infected with Cowdria ruminantium.

A 3-fold increase in the numbers of Lyt-2+ T cells in the circulating blood of mice infected and re-infected with the Welgevonden stock of Cowdria ruminantium, as determined by flow cytometry, is supportive evidence that immunity in heartwater is cell-mediated. The rise in Lyt-2+ cells only after re-infection of the mice is further evidence that the development of immunity in heartwater is dependent on the unhindered and adequate replication of C. ruminantium.     

An improved ferritin assay for canine sera

An improved serum ferritin assay for canine serum has been developed. It uses two monoclonal antibodies in a sandwich arrangement. Serum ferritin can be determined on undiluted canine sera with this assay. The recovery of ferritin added to canine serum ranged from 98 to 106%, the within-assay coefficient of variability was 3.3 to 4.5%, and the assay-to-assay variability was 9.8 to 10.2%. Serum ferritin from 61 apparently healthy dogs had a geometric mean of 252 ng/ml, with a range of 80 ng/ml to 800 ng/ml.     

The relationship between the pharmacokinetics of intravenous cismethrin and bioresmethrin and their mammalian toxicity

The distribution and excretion of [¹⁴C]alcohol-labeled cismethrin and bioresmethrin was determined after intravenous administration to rats. Initially the label distribution of both isomers was similar, but differences occurred at later times mainly due to the retention of 5-benzyl-3-furylcarboxylic acid, a metabolite of bioresmethrin, in high concentration in the blood. Retention of this metabolite accounted for the slower excretion of bioresmethrin label compared to cismethrin. After administration of either isomer, parent pyrethroid was rapidly cleared from the blood and liver, and both isomers rapidly entered the central nervous system reaching peak concentrations within 2–5 min. Brain cismethrin concentrations exceeding 3.5 nmol/g were associated only with animals showing tremors. These levels of cismethrin are maintained for up to 30 min but bioresmethrin was depleted more rapidly possibly due to brain metabolism. It is concluded that the low toxicity of bioresmethrin is possibly due to the inability of this isomer to interact with the site of action in the central nervous system and not, as previously suggested, primarily because of more rapid metabolism in the liver.     

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.     

Predicting the age of fish using general and generalized linear models of biometric data: A case study of two estuarine finfish from New South Wales, Australia

Direct ageing of fish can be a laborious and expensive task when age estimates from a large population are required, and often involves a degree of subjectivity. This study examined the application of general and generalized linear models that predict the age of fish from a range of efficiently and objectively measured covariates. The data sampled were from yellowfin bream (Acanthopagrus australis (Sparidae) (Owen, 1853)) and sand whiting (Sillago ciliata (Sillaginidae) Cuvier, 1829) populations from New South Wales, Australia. The covariates evaluated in the models were fish length, otolith weight, sex and location (the estuary from which the fish were sampled). Akaike Information Criteria were used for model selection and residual plots of the final models revealed a satisfactory fit to the observations. The best fitting model for each species included all covariates. An additional investigation considered whether general and generalized linear models that predict age from two different categories of biometric information outperform age-length keys with respect to subsequent estimates of total mortality from catch-curve analysis. The two categories of biometric information differed in the ease and cost with which the information could be collected. The first category only included fish length and location as covariates, whilst the second category also included otolith weight and sex. It was found that traditional age-length keys outperformed the predictive models that estimated age from only fish length and location, because the results from the models were prone to significant bias. However, when otolith weight and sex were added as covariates to the predictive models, some of them, including a generalized linear model with a Poisson-distributed response variable, performed similarly to the age-length key. Given that otolith weight and the sex of fish are cheaper to quantify than age from a sectioned otolith in many situations, general or generalized linear models may represent a cheaper and faster method of estimating mortality compared to age-length keys. Such models can also easily incorporate the influence of spatial, temporal and demographic variation.     

Resource patch array use by two squirrel species in an agricultural landscape

Eastern grey squirrels (Sciurus carolinensis) and North American red squirrels (Tamiasciurus hudsonicus) were studied among wooded patches within an agricultural mosaic. Fifteen sites south of Ottawa, Canada, with differing landscape and local features were censused using tracking boards placed in a woods or wooded fencerow. Regression analyses of landscape compositional and physiognomic variables within a 1-km radius isolated the best predictors of grey and red squirrel abundance and activity. Grey squirrels were found in both small woods and fencerows in farm landscapes but were not found in large woods. A polynomial regression of wooded patch size explained 79% of the variance in grey squirrel abundance. Grey squirrel activity was correlated with the percent cover of soybeans in the landscape. Red squirrels were found in fencerows, small and large woods; activity was correlated with the percent cover of both woods and corn crop in the surrounding landscape. These results indicate that distributions of both species are influenced by multiple landscape elements, but that grey squirrels may rely on fragmented agricultural landscapes whereas red squirrels make more use of both native woodland and altered landscapes.     

Metal contamination from open-cast copper and sulphur mining in southeast Ireland

The metal content was determined in soil and vegetation around the eastern zone of the disused sulphur and copper mines at Avoca (southeast Ireland). A 250 m × 250 m grid survey was conducted covering a total area of 4 km². The mining belt was in the centre of the investigated site and was sampled separately. Iron, Zn, Cu and Cd concentrations in soil and plant material were determined, in addition to soil organic matter content and soil pH. Soil samples taken from the spoil showed elevated Cu and Cd concentrations, and low soil pH and organic matter content. At least 50 per cent of the agricultural sites examined around the East Avoca Mine showed elevated soil Cu and Cd concentrations, whereas Zn and Fe soil concentrations were within the range for unpolluted soils. None of the vegetation analysed had elevated Zn, Cd, Fe or Cu concentrations. Overall there was no indication of serious metal contamination in either soils or vegetation surrounding the mine in spite of extensive dust deposition during open-pit mining activity 15–30 years ago. However, a plume of elevated Cu in the soil was identified, which originated from a large spoil heap and had been spread for some 2·0 km by the prevailing wind.