Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.     

Familial Congenital Methemoglobinemia in Pomeranian Dogs Caused by a Missense Variant in the NADH‐Cytochrome B5 Reductase Gene

Background

In veterinary medicine, congenital methemoglobinemia associated with nicotinamide adenine dinucleotide (NADH)‐cytochrome b5 reductase (b5R) deficiency is rare. It has been reported in several breeds of dogs, but little information is available about its etiology.

Objectives

To analyze the NADH‐cytochrome b5 reductase gene, CYB5R3, in a Pomeranian dog family with methemoglobinemia suspected to be caused by congenital b5R deficiency.

Animals

Three Pomeranian dogs from a family with methemoglobinemia were analyzed. Five healthy beagles and 5 nonrelated Pomeranian dogs without methemoglobinemia were used as controls.

Methods

Methemoglobin concentration, b5R activity, and reduced glutathione (GSH) concentration were measured, and a turbidity index was used to evaluate Heinz body formation. The CYB5R3 genes of the affected dog and healthy dogs were analyzed by direct sequencing.

Results

Methemoglobin concentrations in erythrocytes of the affected dogs were remarkably higher than those of the control dogs. The b5R activity of the affected dogs was notably lower than that of the control dogs. DNA sequencing indicated that this Pomeranian family carried a CYB5R3 gene missense variant (ATC→CTC at codon 194) that resulted in the replacement of isoleucine (Ile) by leucine (Leu).

Conclusions and Clinical Importance

This dog family had familial congenital methemoglobinemia caused by b5R deficiency, which resulted from a nonsynonymous variant in the CYB5R3 gene. This variation (c.580A>C) led to an amino acid substitution (p.Ile194Leu), and Ile194 was located in the proximal region of the NADH‐binding motif. Our data suggested that this variant in the canine CYB5R3 gene would affect function of the b5R in erythrocytes.     

Glycine max and Glycine soja are capable of cold acclimation

Soybean has been considered a cold intolerant species; based largely upon seed germination and soil emergent evaluations. This study reports a distinct acquisition of cold tolerance, in seedlings, following short acclimation periods. Diversity in cold responses was assessed in eight cultivars of Glycine max and six accessions of G. soja. All varieties of soybean significantly increased in freezing tolerance following acclimation. This study indicates soybean seedlings are indeed capable of sensing cold and acquiring cold tolerance. Germination rates after cold imbibition were negatively correlated with maturity group, but positively correlated with cold acclimation potential in G. soja. Seed fatty acid composition was varied between the species, with Glycine soja accessions containing about 2‐times more linolenic acid (18:3) than G. max. Furthermore, high levels of linoleic acid (18:2) in seeds were positively correlated with germination rates following cold imbibition in G. soja only. We suggest that domestication has not impacted the overall ability of soybean to cold acclimate at the seedling stage and that there is little variation within the domesticated species for ability to cold acclimate. Thus, this brief comparative study reduces the enthusiasm for the “wild” species as an additional source of genetic diversity for cold tolerance.     

Hemorrhagic necrotizing splenitis in a slaughter pig infected with Arcanobacterium species

A 6-month-old barrow presented with lethargy, inappetence and dysstasia. At necropsy, multiple coalescing hemorrhagic foci were detected in the margins of the spleen. Gram-positive bacilli were isolated from the spleen, kidney, muscle and liver. Comparative 16S rDNA gene sequencing analysis of the isolates (TO16177) revealed that they would be the same species of unpublished Arcanobacterium species strain HJ57-14E (accession no. gi 18873551) (99.7% similarity based on a comparison of 675 bp). Histologic examination of the splenic tissue sections revealed extensive necrosis and inflammation, and gram-positive bacilli were discernible. Multifocal necrosis was also detected in the liver. Immunohistochemically, the isolates were cross-reacted with polyclonal antibodies against Arcanobacterium pyogenes and Actinomyces naeslundii, and the reaction was strong for the latter. Similar reactions were found in the suppurative lesions of the tonsil, and occasionally in the spleen and lymph nodes. The present results indicate that the unpublished Arcanobacterium species induced multiple organ failure accompanied by acute hemorrhagic necrotizing splenitis in this growing-finishing pig.     

Adsorption of Cr(VI) ion on synthetic hydrated oxides of iron

Abstract

The adsorption of Cr(VI) ion on synthetic hydrated oxides of iron (goethite (α-FeOOH) and lepidocrocite (γ-FeOOH)) with different crystallinities was studied in order to remove the Cr(VI) ion from polluted water. At pH 4.5, the amounts of the adsorbed Cr(V1) ion on goethite and lepidocrocite were 51.9 and 62.2 mmol g^?, respectively. Probably, this is due to the fact that the structure of lepidocrocite is more open than that of goethite. Lepidocrocite, therefore, is more effective for the removal of Cr( V1) ion from Cr(Vl)-contaminated and acidic groundwater and sewage.     

The influence of phosphate fertilizer application levels and cultivars on cadmium uptake by Komatsuna (Brassica rapa L. var. perviridis)

Cadmium (Cd) is a common impurity in phosphate fertilizers and application of phosphate fertilizer may contribute to soil Cd accumulation. Changes in Cd burdens to agricultural soils and the potential for plant Cd accumulation resulting from fertilizer input were investigated in this study. A field experiment was conducted on Haplaquept to investigate the influence of calcium superphosphate on extractable and total soil Cd and on growth and Cd uptake of different Komatsuna (Brassica rapa L. var. perviridis) cultivars. Four cultivars of Komatsuna were grown on the soil and harvested after 60 days. The superphosphate application increased total soil Cd from 2.51 to 2.75?mg?kg^?1, 0.1?mol?L^?1 hydrochloric acid (HCl) extractable Cd from 1.48 to 1.55?mg?kg^?1, 0.01?mol?L^?1 HCl extractable Cd from 0.043 to 0.046?mg?kg^?1 and water extractable Cd from 0.0057 to 0.0077?mg?kg^?1. Cd input reached 5.68?g?ha^–1 at a rate of 240?kg?ha^–1 superphosphate fertilizer application. Superphosphate affected dry-matter yield of leaves to different degrees in each cultivar. ‘Nakamachi’ produced the highest yield in 2008 and ‘Hamami No. 2’ in 2009. Compared with the control (no phosphate fertilizer), application of superphosphate at a rate of 240?kg?ha^–1 increased the Cd concentration in dry leaves by 0.14?mg?kg^?1 in ‘Maruha’, 1.03?mg?kg^?1 in ‘Nakamachi’, 0.63?mg?kg^?1 in ‘SC8-007’ in 2008, and by 0.19?mg?kg^?1 in Maruha’, 0.17?mg?kg^?1 in ‘Hamami No. 2’, while it decreased by 0.27?mg?kg^?1 in ‘Nakamachi’ in 2009. Field experiments in two years demonstrated that applications of different levels of calcium superphosphate did not influence Cd concentration in soil and Komatsuna significantly. However, there was a significant difference in Cd concentration of fresh and dry Komatsuna leaves among four cultivars in 2008 and 2009. The highest Cd concentration was found in the ‘Nakamachi’ cultivar (2.14?mg?kg^?1 in 2008 and 1.91?mg?kg^?1 in 2009). The lowest Cd concentration was observed in the ‘Maruha’ cultivar (1.51?mg?kg^?1?dry weight (DW)) in 2008 and in the ‘Hamami No. 2’ cultivar (1.56?mg?kg^?1?DW) in 2009. A decreasing trend in Cd concentration was found in ‘Nakamachi’, followed by ‘SC8-007’, ‘Hamami No. 2’ and ‘Maruha’ successively. It is necessary to consider a low-uptake cultivar for growing in a Cd polluted soil. In these two years’ results, ‘Maruha’ cultivar was the lowest Cd uptake cultivar compared to the others.     

Yield estimation of 1-year-old asparagus grown using rootstock-planting forcing culture

Rootstock-planting forcing culture was developed in asparagus to harvest spears even during the seasons when the plants become dormant, but the demand for them high. In this study, cumulative hours during which the air temperature remained lower than 5°C, i.e. chilling hours (CHs), were calculated to determine dormancy breakage for asparagus cultures. We also measured CIELab colour values for cut stems immediately before rootstock digging, and determined whether they could be substituted and/or compensated for CHs while evaluating asparagus plant productivity in different low-temperature backgrounds, and obtained regression equations for yield estimation. Asparagus seedlings were cultivated in seven different regions across Japan and brought to the study site for harvesting. Our regression equation based on CHs and rootstock weight for yield estimation had relatively high fitness (adjusted R² = 0.5795). The colour values of cut stalks at rootstock digging can also be used to evaluate their productivity. These values can be useful in regions where CHs cannot be determined, although their effectiveness was slightly lower than that of CHs of areas adjacent to the study sites.     

A Generalized Two-Dimensional Gaussian Model of Disease Foci of Head Blight of Wheat Caused by Gibberella zeae

ABSTRACT A generalized two-dimensional Gaussian model is proposed to describe disease foci of head blight of wheat in plots (100 to 2,500 m(2)) originating from small areas (1 to 16 m(2)) inoculated with Gibberella zeae-colonized corn kernels. These anisotropic, asymmetrical foci arose from ascospores produced in perithecia. The model is Z = exp[-(AX(2) + BY(2) + CXY + DX + EY + F)], in which Z = the incidence of seed or spikelet infection at point (X,Y) located in the plot, exp = the exponential function, X = the abscissa or spatial coordinate of the point along a given axis (approximately parallel to the average wind vector during the period of spore release in these experiments), Y = the ordinate or spatial coordinate of the point along the axis perpendicular to the X axis (approximately perpendicular to the wind direction in these experiments), A and B = the quadratic coefficients of the second-order polynomial AX(2) + BY(2) + CXY + DX + EY + F, C = the bilinear coefficient, D and E = the linear coefficients, and exp(-F) = the incidence of seed or spikelet infection at the focus peak in which X = 0 and Y = 0. The generalized two-dimensional Gaussian model was tested on data from a circular or isotropic focus, an elliptical or anisotropic focus with two axes of symmetry, and two anisotropic foci with one and zero axis of symmetry. Its goodness-of-fit (r(2) and adjusted r(2)) was compared with the inverse power, modified inverse power, exponential, and classical Gaussian models. Submodels using only the linear terms, only the quadratic terms, or combinations selected from stepwise regression procedures using various probabilities to enter and to stay and a procedure maximizing the adjusted r (2) were also considered. Spatial analysis of the residuals was performed using Geary's c coefficient at the first distance class. For the circular and elliptical foci, our model provided a fit similar to the modified inverse power and exponential models. However, for anisotropic foci with one or zero axis of symmetry arising from ascospores influenced by wind direction, the generalized two-dimensional Gaussian model provided a better fit. For these anisotropic foci, the linear term X but not the quadratic term X(2) was generally retained in the model, indicating an exponential gradient in the direction parallel to the wind. In all models, the quadratic term Y(2) was retained, along with Y in some cases, indicating that the gradient in the direction roughly perpendicular to the wind was Gaussian or Gaussian-exponential in shape. The bilinear term XY provided an indication of the orientation of the focus in relation to the axes of the sampling grid. This model has the versatility and parameters (quadratic, bilinear, and linear) to better describe the anisotropy of foci from wind-dispersed spores.