Western blot detection of PrP Sc in archived paraffin-embedded brainstem from scrapie-affected sheep

Scrapie is a naturally occurring fatal neurodegenerative disease of adult sheep and goats, one of a group of mammalian diseases known as transmissible spongiform encephalopathies (TSE) or prion diseases. Immunoassays that identify disease-associated prion protein (PrP Sc) are integral to the diagnosis of scrapie and other prion diseases. Results obtained by either immunohistochemistry (IHC) or Western blot (WB) assay are generally adequate for the definitive diagnosis. Approved or accepted methods for WB diagnosis of TSEs requires the use of fresh or frozen nonfixed tissue samples, whereas formalin-fixed, paraffin-embedded tissue is required for the localization of PrP Sc by IHC. Because disparate processing methods are used for these accepted diagnostic techniques, separate tissue samples are collected from the same animal. Occasions arise in which there is either insufficient quantity of tissue available to complete analysis by both techniques or initial tissue processing is incompatible with one of the assays. Also, results between the assays may differ because of the vagaries of sampling, especially in case material that contains moderate-to-low levels of PrP Sc. The present article describes a method to conduct a WB assay from the same paraffin-embedded brainstem sample used for the IHC diagnosis of experimentally induced sheep scrapie.     

The role of wildlife tourism in conservation of endangered species: Implications of safari tourism for conservation of the Arctic fox in Sweden

There are both positive and negative impacts on wildlife associated with wildlife tourism. In Sweden, the endangered Arctic fox is subject to a growing tourist interest. In the Helags mountain region there are guided Arctic fox safari tours that provide visitors with information about the Arctic fox. A survey of five separate groups of visitors in the region revealed that knowledge about the status of Arctic foxes and awareness of the behavioral guidelines for Arctic fox encounters improved after participation in a safari tour and with increasing Arctic fox interaction. We propose a schematic model summarizing the diverse ways in which wildlife tourism affects wildlife and their relative importance for conservation. The Arctic fox population in Sweden is small and sensitive to disturbance, but the positive impacts of Arctic fox tourism seem to compensate for the negative and contribute to their conservation under the current level of tourism pressure.     

Changes of Some Components of Precipitation in East Germany after the Unification

A network of 15 sites of precipitation monitoring was built-up to characterize the spatial and temporal changes of pollutants over East Germany. Rain water samples were analysed and characterized regarding their acidity, conductivity, main water soluble components (Cl^-, NO₃ ^-, SO₄ ^2-, Na⁺, NH₄ ⁺, K⁺, Ca²⁺, Mg²⁺), volume and the meteorological parameters. Dependent on the changes of emission of pollutants, resulting from the reformation of industry, the decrease of stock farming, and increase of traffic (by the factor of ≈1.5) in East Germany, a drop could be observed with the sulphate and calcium concentrations in the precipitation by 20 to 50 and 30 to 70%, respectively. An increase could not be ascertained with the nitrate concentration and the acidity.     

Antibacterial properties of silk fibroin/chitosan blend films loaded with plant extract

The silk fibroin (SF)/chitosan (CHI) blend films were prepared by dissolving them in formic acid. The morphology of the films was examined by scanning electron microscopy (SEM). The roughness of the membranes was determined by atomic force microscopy (AFM). These films were treated with the extracts of Pistacia terebinthus, Pistacia lentiscus, and Hypericum empetrifolium. Folin-Ciocalteu assay was used to determine the amount of total phenols adsorbed on these blend films. The antibacterial properties of films were tested using disc diffusion and turbidity measurement methods against Escherichia coli and Staphylococcus epidermidis. The release of adsorbed phenolics from the film surface resulted in antibacterial properties.     

Evaluation of EDTA hematology tubes for collection of blood samples for tests of secondary hemostasis in dogs

OBJECTIVE: To evaluate the use of EDTA tubes for collection of blood samples for assays of secondary hemostasis in dogs. ANIMALS: 108 dogs of various ages, breeds, and sexes (19 healthy and 89 with abnormalities of secondary hemostasis). PROCEDURES: Blood samples were collected via cephalic venipuncture and transferred to sodium citrate tubes and EDTA tubes. Plasma was harvested from each type of tube for assays of concentrations of fibrinogen and D-dimer as well as prothrombin time, activated partial thromboplastin time, and antithrombin activity. Intra-assay and interassay precision and correlation coefficients for all hemostatic tests were calculated for each type of plasma sample. The effect of storage conditions on assay results for the 2 types of plasma samples was also evaluated. RESULTS: Results of hemostatic tests were highly correlated between citrated and EDTA-treated plasma samples. Intra-assay imprecision for all hemostatic tests with the exception of D-dimer concentration was < 10% for both citrated and EDTA-treated plasma samples; interassay imprecision was higher for EDTA-treated versus citrated plasma samples. Storage of plasma samples for 1 hour did not result in significantly different assay results for either type of plasma sample, but storage for 2 hours significantly affected values for EDTA-treated plasma samples. CONCLUSIONS AND CLINICAL RELEVANCE: Although evaluation of the sensitivity and specificity of hemostatic tests that use EDTA-treated plasma samples is required, EDTA may be a suitable alternative to sodium citrate as an anticoagulant for use in hemostatic testing in conditions in which tests could be performed within 1 hour after sample collection.     

Plasma levels of insulin,glucagon and glucagon-like peptide in salmonids of different weights

Plasma levels of insulin in rainbow trout,Oncorhynchus mykiss, Atlantic salmon,Salmo salar, and Pacific coho salmon,Oncorhynchus kisutch and plasma circulating levels of glucagon and glucagon-like peptide, in rainbow trout and Atlantic salmon, were measured by homologous radioimmunoassays. Hormonal levels were compared against the average body weight of the same group of fish. Plasma insulin levels were significantly correlated (r=0.56, 0.46 and 0.42 respectively) with body weight in all three salmonid species. Moreover, rainbow trout from fast-growing families had significantly higher (p<0.005) plasma insulin levels than did fish from slow-growing families. Plasma titres of glucagon and glucagon-like peptide were always lower than insulin titres and did not correlate with body weight.Reported in part at Satellite Symposium on Applications of Comparative Endocrinology to Fish Culture, Almunecar, Spain (Sundby, 1989).     

Demand and effect of potassium,magnesium, and calcium chlorides on hemolymph parameters,immune-related gene expression,and growth of Litopenaeus vannamei,Boone (1931) under biofloc technology

The ionic composition of culture water may be a more important limiting factor than the salinity itself and may lead to osmotic stress which may influence growth and survival of shrimp culture. The uptake rate and the effect of magnesium chloride (MgCl₂), calcium chloride (CaCl₂), and potassium chloride (KCl) salts in juveniles of whiteleg shrimp (Litopenaeus vannamei) cultured with biofloc technology (BFT) was evaluated for 62 days in seawater (30 practical salinity unity). Five treatments were analyzed in triplicate: T1; control (water exchange rate of 5% daily), T2; adding CaCl₂?+?MgCl₂?+?KCl, T3; adding MgCl₂?+?KCl, T4; adding CaCl₂?+?KCl, and T5; adding CaCl₂?+?MgCl₂. Mineral salts were added to water and the response of experimental parameters: physicochemical variables of water quality, osmotic pressure, total hemocyte count, glutathione peroxidase gene expression, superoxide dismutase, and zootechnical variables of the shrimp was assessed. The uptake of single chloride salts (CaCl₂, MgCl₂, and KCl) by shrimp varied as a function of the concentrations and the ratio of these three ions. Magnesium deficiencies in the culture medium increased CaCl₂ and KCl uptake without showing gene expression of SOD and GPx. The best survival rate was obtained by adding the three ions (T2) and the control (T1, water exchange of 5%). We concluded that L. vannamei can be maintained by the addition of essential chloride ions in BFT without water replacement.

     

Morphological, histological, and ultrastructural studies of the ovary of the cattle-tick Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae)

This study presents the morphology of the ovary, as well as the dynamics of the vitellogenesis process in oocytes of the cattle-tick Boophilus microplus. The ovary of these individuals is of the panoistic type; therefore, it lacks nurse cells. This organ consists of a single tubular structure, continuous, and composed of a lumen delimitated by a wall of small epithelial cells with rounded nuclei. In this tick species, the oocytes were classified into six stages varying from I to VI and according to: cytoplasm appearance and presence of the germ vesicle, yolk granules, and chorion. Oocytes of various sizes and at different developmental stages remain attached to the ovary through a cellular pedicel until completing stage V. Afterwards, they are liberated into the lumen and from there to the exterior. Some oocytes (classified as type VI) showed an atypical appearance indicating that some of the cellular components would be undergoing a degenerative process and/or reabsorption.     

Arsenic and chromium resistance mechanisms in the Micrococcus luteus group

High arsenic (As) and chromium (Cr) concentrations are currently receiving attention because of their negative effects on the environment and human health. Microorganisms inhabiting contaminated environments have developed resistance mechanisms against the toxicity of these pollutants. Indeed, members of the bacterial genus Micrococcus have been isolated from different toxic metal-contaminated environments; however, knowledge concerning its resistance mechanisms to As and Cr toxicity remains lim...