Non-structural plum pox potyvirus proteins detected by immunogold labelling

Plum pox potyvirus (PPV) induces in infected Nicotiana clevelandii cells characteristic crystalline inclusions known as nuclear inclusions (NI) when located in the nucleus and as dense material (Dm) when located in the cytoplasm. Crystalline inclusions contain protease (NIa) and RNA-dependent RNA polymerase (NIb) proteins. It is now well established for all potyviruses that cylindrical inclusions contain CI helicase ATPase protein (Martin et al., 1992). The intracellular location of other non-structural PPV proteins remains unknown. Using Escherichia coli expression vectors, specific antibodies were obtained against P1, P3, 6K2 and NIb PPV proteins for which antibodies were not yet available. As expected, NIb antiserum labelled crystalline inclusions. P1, P3 and 6K2 proteins were present in both types of crystalline inclusions found in the nucleus and in the cytoplasm of PPV-infected leaves of N. clevelandii, suggesting that nuclear inclusions and dense material were composed of the same proteins. This composition is discussed.     

Immunohistochemical differentiation of reactive from malignant mesothelium as a diagnostic aid in canine pericardial disease

Objectives

To develop a provisional immunohistochemistry panel for distinguishing reactive pericardium, atypical mesothelial proliferation and mesothelioma in dogs.

Materials and Methods

Archived pericardial biopsies were subject to haematoxylin and eosin staining, immunohistochemistry for cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Samples were scored for intensity and number of cells stained.

Results

Ten biopsies of reactive mesothelium, 17 of atypical mesothelial proliferation, 26 of mesothelioma and five of normal pericardium were identified on the basis of haematoxylin and eosin staining. Cytokeratin and vimentin were expressed in all biopsies, confirming mesothelial origin. Normal pericardial samples had the lowest scores for insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Mesothelioma and atypical proliferative samples were similar to each other, with higher scores for insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 than the reactive samples. Desmin staining was variable. Insulin‐like growth factor II mRNA‐binding protein 3 was the best to distinguish between disease groups.

Clinical Significance

An immunohistochemistry panel of cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 could provide superior information compared with haematoxylin and eosin staining alone in the diagnosis of cases of mesothelial proliferation in canine pericardium, but further validation is warranted.     

Knowledge,attitudes and practices towards <Emphasis Type="Italic">cystic echinococcosis</Emphasis> in livestock among selected pastoral and agro-pastoral communities in Uganda

A cross-sectional study was done from March 2013 to May 2014 to assess knowledge, attitudes, and practices towards cystic echinococcosis (CE) or hydatidosis among selected pastoral and agro-pastoral communities in Uganda. A structured questionnaire was administered to 381 respondents. Multivariate logistic regression analysis was done to find the relationship between knowledge about CE and factors such as age, sex, and level of education across all regions. The odds ratio and confidence interval were used to determine the difference in responses across regions. It was shown that age above 36 years was significantly (p < 0.001) associated with awareness about CE in livestock. Likewise, uneducated (p < 0.0001) and agro-pastoralists (p = 0.01) were significantly less knowledgeable than the educated and pastoralists across all regions. The overall knowledge towards CE in livestock was low 17.8% (95% CI = 14.0–21.6). Dog ownership was high and they never dewormed their freely roaming dogs. Dogs shared water with livestock. In conclusion, knowledge about CE in livestock was low across all regions. Therefore, public health education and formulation of policies towards its control by the relevant stakeholders should be done. Also, the true prevalence of CE in livestock needs to be done so that the magnitude and its public health significance are elucidated.     

Colorimetric determination of arprinocid in feed.

An analytical method has been developed for the determination of arprinocid (9-(2-chloro-6-fluorophenylmethyl)-9H-purin-6-amine) in feed, based upon measurement of the absorbance of the diazo chromophore formed from a product of zinc reduction of the drug in acidic solution. The analyte is extracted from the feed into chloroform in the presence of a pH 7 phosphate buffer and isolated by adsorption chromatography on alumina, followed by partitioning between hexane and 0.15M HCl. The reduction product in the aqueous phase is then treated for colorimetric measurement. This procedure has been applied to determining 0.0010--0.0080% arprinocid in feed with a precision of less than 5% relative standard deviation near the middle of this concentration range. Of 32 feed additives examined, only zoalene and sulfamethazine were serious interferences. A study and discussion of several factors, e.g., reaction time, pH, and amount of zinc metal, that affect the analytical reactions are also included.     

Carbon loss from roots of wheat cultivars

The amounts of organic materials released into soil from roots during the first 4 weeks of growth were determined for 11 cultivars of wheat (Triticum aestivum L.). Carbon loss from roots was measured by supplying ¹⁴CO₂ continuously to the shoots and measuring the ¹⁴C content of the roots, root-free soil, water-soluble material and CO₂ flushed from the root chamber. Six cultivars were compared in each of two experiments, with the cultivar Condor common to both experiments. There were no significant differences between cultivars, relative to Condor, for ¹⁴C activity present in soil, roots, water-soluble material or rhizosphere CO₂. There was a significant difference between cultivars in experiment 1, but not in experiment 2, for the variate log₁₀ (¹⁴C lost from roots: ¹⁴C translocated to roots).There was evidence that a reduction in growth temperature, within the range 10–15°C, increased carbon loss from wheat roots into the rhizosphere.     

Decomposition of algal cells and components and their stabilization through complexing with model humic acid-type phenolic polymers

Axenic cultures of Anacystis, Microcoleus, Plectonema and Synechococcus isolated from Greenfield sandy loam and of Anabaena flos-aquae, Nostoc muscorum and Chlorella pyrenoidosa from other sources were cultured under light and constant aeration and with [U-¹⁴C]-glucose in the nutrient medium. Whole cells, cell walls, cytoplasm and extracellular polysaccharides of selected species readily decomposed in the soil and after 22 weeks between 61 and 81% of the added C had evolved as CO₂. Complexing of cell wall and cytoplasmic preparations from A. flos-aquae and N. muscorum with model humic acid-type phenolic polymers reduced decomposition of the cell walls by 40% and of the cytoplasm by 70%. Over 50% of the residual 14C activity in the soil amended with whole algal cells remained in the 0.5% NaOH-extracted soil. With exception of Microcoleus sp. more of the residual ¹⁴C from cell walls, cytoplasm and polysaccharide fractions was present in the humic acid or fulvic acid fractions.     

Effect of montmorillonite and humate on growth and metabolic activity of some actinomycetes

In well-aerated culture solutions Ca-montmorillonite at 0.25% concentration markedly accelerated and increased growth, glucose consumption and CO₂ evolution by various Streptomyces, Micromonospora and Nocardia species. The montmorillonite was a little more active than Ca-humate and was usually still but somewhat less effective when confined to dialysis tubing. Ca-exchange resin, Na₂SiO₃ and finely powdered CaCO₃ exerted very little or no effect. In many cultures the relation of glucose consumption to biomass formation indicated a more efficient use of the glucose C for cell synthesis in the presence of clay. In other cultures the greater biomass formation was associated with a more rapid and complete utilization of the glucose present.