A multicenter case-control study of risk factors for equine protozoal myeloencephalitis

OBJECTIVE: To identify risk factors for equine protozoal myeloencephalitis (EPM) among horses examined at 11 equine referral hospitals. DESIGN: Case-control study. ANIMALS: 183 horses with EPM, 297 horses with neurologic disease other than EPM (neurologic controls), and 168 horses with non-neurologic diseases (non-neurologic controls) examined at 11 equine referral hospitals in the United States. PROCEDURES: A study data form was completed for all horses. Data were compared between the case group and each of the control groups by means of bivariate and multivariate polytomous logistic regression. RESULTS: Relative to neurologic control horses, case horses were more likely to be > or = 2 years old and to have a history of cats residing on the premises. Relative to non-neurologic control horses, case horses were more likely to be used for racing or Western performance. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that cats may play a role in the natural epidemiology of EPM, that the disease is less common among horses < 2 years of age relative to other neurologic diseases, and that horses used for particular types of competition may have an increased risk of developing EPM.     

Development of a gene silencing DNA vector derived from a broad host range geminivirus

Background  

Gene silencing is proving to be a powerful tool for genetic, developmental, and physiological analyses. The use of viral induced gene silencing (VIGS) offers advantages to transgenic approaches as it can be potentially applied to non-model systems for which transgenic techniques are not readily available. However, many VIGS vectors are derived from Gemini viruses that have limited host ranges. We present a new, unipartite vector that is derived from a curtovirus that has a broad host range and will be amenable to use in many non-model systems.     

Flower greening in phytoplasma-infected Hydrangea macrophylla grown under different shading conditions

To determine the effect of light intensity on flower greening, the Japanese hydrangea phyllody (JHP) phytoplasma-infected hydrangea cultivars ‘Midori’, ‘Libelle’, ‘Rosea’ and ‘Madame E. Mouillere’ plants were grown under different shade conditions. In the first-year experiment, the results indicate that the flowers of the JHP-phytoplasma-infected hydrangea become green under shaded conditions (70% and 49% sunlight intensities). On the other hand, under full sunlight intensity (100% sunlight intensity), the flowers of ‘Midori’, ‘Rosea’, and ‘Libelle’ plants were blue, pink or white. To calculate the percentage of flower greening, inflorescences of these plants were separated and divided into individual flowers, and classified into four types by green-area ratio, calculated using Adobe Photoshop. Under shading with one sheet of cheesecloth (70% sunlight intensity), the inflorescences of ‘Midori’, ‘Libelle’ and ‘Madame E. Mouillere’ plants were composed of more than 40% completely green flowers (0.8 ≦ green-area ratio), whereas those of ‘Rosea’ plant had 0% completely green flowers. Under shading with two sheets of cheesecloth (49% sunlight intensity), the inflorescences of ‘Midori’, ‘Libelle’ and ‘Madame E. Mouillere’ plants had more than 75% completely green flowers; ‘Rosea’ plants had 28%. In the second-year experiment, under full sunlight intensity, ‘Midori’ plants had four types of flower depending on their green-area ratio, namely, completely blue or pink, pink-green, greenish and completely green flowers. Under shading with two sheets of cheesecloth, ‘Midori’ plants had more than 90% completely green flowers. The JHP-phytoplasma could not be identified by PCR analysis in flowers with a green-area ratio = 0 (completely blue/pink/white flowers). On the other hand, in flowers with a green-area ratio > 0, the JHP-phytoplasma was detected by PCR analysis. Thus, we conclude that shading enhances flower greening in hydrangea by increasing the JHP-phytoplasma concentration in the flowers.     

Shifts in bacterial community structure associated with inputs of low molecular weight carbon compounds to soil

Low molecular weight carbon (C) substrates are major drivers of bacterial activity and diversity in the soil environment. However, it is not well understood how specific low molecular weight C compounds, which are frequently found in root exudates and litter leachates, influence bacterial community structure or if there are specific groups of soil bacteria that preferentially respond to these C inputs. To address these knowledge gaps, we added three simple C substrates representative of common root exudate compounds (glucose, glycine, and citric acid) to microcosms containing three distinct soils from a grassland, hardwood forest, and coniferous forest. CO₂ production was assessed over a 24 h incubation period and, at the end of the incubation, DNA was extracted from the samples for assessment of bacterial community structure via bar-coded pyrosequencing of the 16S rRNA gene. All three C substrates significantly increased CO₂ production in all soils; however, there was no relationship between the magnitude of the increase in CO₂ production and the shift in bacterial community composition. All three substrates had significant effects on overall community structure with the changes primarily driven by relative increases in β-Proteobacteria, γ-Proteobacteria, and Actinobacteria. Citric acid additions had a particularly strong influence on bacterial communities, producing a 2-5-fold increase in the relative abundance of the β-Proteobacteria subphylum. These results suggest that although community-level responses to substrate additions vary depending on the substrate and soil in question, there are specific bacterial taxa that preferentially respond to the substrate additions across soil types.     

Variations in equid SLC11A1 (NRAMP1) genes and associations with Rhodococcus equi pneumonia in horses

Rhodococcus equi is an important intracellular pathogen of horses, most commonly causing chronic, suppurative bronchopneumonia in foals. Although most foals likely are exposed to environmental R. equi within the 1st few days of life, only some develop R. equi pneumonia, and the basis of differences in susceptibility among foals currently is unknown. In this study, we investigated solute carrier family 11 member 1 (SLC11A1) gene sequences in the 5' untranslated region, exon 1, and a portion of intron 1 for variations in 3 equid species (horse, donkey, zebra) and compared variants within 3 independent horse breeding farms for associations with R. equi pneumonia by use of an age-matched case-control design. Seven novel variants in the 5'untranslated region were identified as specific for one or both of the non-horse equid species sampled. In addition, a single novel horse variant in the 5'untranslated region, -57C/T, was identified in 4 breeds. The -57C/T variant was found on 2 of the 3 farms with endemic R. equi pneumonia, representing 2 different horse breeds. Significant allelic and genotypic associations with susceptibility to R. equi pneumonia were observed for the -57C/T variant in foals from these farms. Although the functional impact of this novel variant remains to be determined, this study represents an important step in our understanding of natural resistance to R. equi foal pneumonia and other intracellular bacterial diseases affecting equids.     

A mitochondrial pyruvate carrier required for pyruvate uptake in yeast, Drosophila, and humans

Pyruvate constitutes a critical branch point in cellular carbon metabolism. We have identified two proteins, Mpc1 and Mpc2, as essential for mitochondrial pyruvate transport in yeast, Drosophila, and humans. Mpc1 and Mpc2 associate to form an ~150-kilodalton complex in the inner mitochondrial membrane. Yeast and Drosophila mutants lacking MPC1 display impaired pyruvate metabolism, with an accumulation of upstream metabolites and a depletion of tricarboxylic acid cycle intermediates. Loss of yeast Mpc1 results in defective mitochondrial pyruvate uptake, and silencing of MPC1 or MPC2 in mammalian cells impairs pyruvate oxidation. A point mutation in MPC1 provides resistance to a known inhibitor of the mitochondrial pyruvate carrier. Human genetic studies of three families with children suffering from lactic acidosis and hyperpyruvatemia revealed a causal locus that mapped to MPC1, changing single amino acids that are conserved throughout eukaryotes. These data demonstrate that Mpc1 and Mpc2 form an essential part of the mitochondrial pyruvate carrier.