Optimization of a Staphylococcus aureus adhesion assay for equine corneocytes

Meticillin-resistant Staphylococcus aureus (MRSA) causes serious skin and soft-tissue infections of humans and animals. Multiple strains of MRSA have been characterized, and one in particular, designated as strain USA 500, causes infections predominantly of horses and the people who work with them. The purpose of this study was to optimize an assay which could subsequently be used to compare the relative avidity of different S. aureus strains for equine corneocytes. Corneocytes were collected from the perineal skin of 10 healthy horses onto adhesive discs. The discs were then incubated at 37°C with an S. aureus field strain at each of three concentrations [10(7), 10(8) and 10(9) colony forming units (CFU)/mL] and for each of three incubation periods (45, 90 and 180 min). After standardized rinsing and staining procedures, discs were examined at ×1,000 magnification and areas containing confluent corneocytes photographed. The percentage of surface area occupied by adherent bacteria was analysed using image processing and analysis software. Significant colour space image processing was required to distinguish bacteria from the ubiquitous melanin granules present within equine corneocytes. Objective and subjective methods were used to determine optimal conditions for specific adherence without introducing confounding factors. A bacterial concentration of 10(8) CFU/mL incubated with corneocytes for 45 min produced maximal bacterial adhesion with the least amount of interbacterial clumping. Future studies should utilize these conditions for optimal assay performance.     

Optimization of lycopene extraction from tomato cell suspension culture by response surface methodology

Radioisotope-labeled lycopene is an important tool for biomedical research but currently is not commercially available. A tomato cell suspension culture system for the production of radioisotope-labeled lycopene was previously developed in our laboratory. In the current study, the goal was to optimize the lycopene extraction efficiency from tomato cell cultures for preparatory high-performance liquid chromatography (HPLC) separation. We employed response surface methodology (RSM), which combines fractional factorial design and a second-degree polynomial model. Tomato cells were homogenized with ethanol, saponified by KOH, and extracted with hexane, and the lycopene content was analyzed by HPLC-PDA. We varied five factors at five levels: ethanol volume (1.33-4 mL/g); homogenization period (0-40 s/g); saturated KOH solution volume (0-0.67 mL/g); hexane volume (1.67-3 mL/g); and vortex period (5-25 s/g). Ridge analysis by SAS suggested that the optimal extraction procedure to extract 1 g of tomato cells was at 1.56 mL of ethanol, 28 s homogenization, 0.29 mL of KOH, 2.49 mL of hexane, and 17.5 s vortex. These optimal conditions predicted by RSM were confirmed to enhance lycopene yield from standardized tomato cell cultures by more than 3-fold.     

Spatial Variability of Agricultural Soil Fertility Parameters in a Gleyic Podzol of Belgium

Precise information about the spatial variability of soil nutrients is essential in developing site-specific management, e.g., variable rate application of nutrients. In this study 5 fields (4 grassland, 1 arable land) on a Gleyic Podzol, each of about 1 ha, were sampled according to a 20 m×20 m grid. On these samples pH, carbon content and nutrient status (P, K, Ca, Mg, and Na) were measured. The variogram analysis of the different soil fertility parameters revealed quite different ranges for these parameters. Carbon content, pH and calcium have high ranges, between 110 and 170 m. On the other hand, ranges for potassium and sodium are much smaller, 60 m and 30 m respectively. For the elements potassium and sodium, high coefficients of variation are found, especially on the grassland fields. These differences in ranges of spatial relationship indicate that the sampling strategy for estimating variability of soil fertility, should be adopted to the different selected soil parameters and the soil use.     

Phosphorus Fractionation in Sediment Cores Collected In 2005 Before and After Onset of an Aphanizomenon flos-aquae Bloom in Upper Klamath Lake, OR, USA

We tested the hypothesis that there would be measurable losses of phosphorus (P) from surficial sediments of Upper Klamath Lake (UKL), Oregon, if sediments were a source of P during an algal bloom. We compared concentrations of total and forms of P at various depths in cores collected before and after the onset of a large Aphanizomenon flos-aquae bloom. Concentrations of inorganic P were determined in extraction solutions of MgCl₂ (1 M, pH 8), citrate-dithionite-bicarbonate, and 1 M HCl. Sediments below 2 cm were dominated by residual P which is defined as total P minus inorganic P. During the study period, data from the top 2-cm of sediment indicated (a) significant decrease in total P concentration, primarily associated with iron oxyhydroxides at one site, and (b) significant increase in total P concentration associated with residual P at a second site. Data from two other sites indicated no net changes in concentrations of total P.     

Conjunctival xanthoma in a blue and gold macaw (Ara ararauna)

A 17-year-old female blue and gold macaw ( Ara ararauna ) presented for evaluation of a discreet, conjunctival mass of the OD. No other abnormalities were found on ophthalmic or physical examination. A heterophilic leukocytosis was present on the complete blood count, and elevated aspartate aminotransferase activity, creatinine kinase activity and cholesterol were present on the plasma biochemistry panel. Surgical removal of the mass was complete and no recurrence has occurred by six months after excision. Reduction of dietary fats was recommended to reduce serum cholesterol levels and reduce the likelihood of future occurrence of xanthomas in this bird.     

Leafy Spurge (Euphorbia esula) Classification Performance Using Hyperspectral and Multispectral Sensors

Two demonstration sites in southeast Idaho were used to extend the scope of remote sensing of leafy spurge research toward investigating coarser scale detection limits. Hyperspectral images were obtained to produce baseline leafy spurge maps, from which spatially and/or spectrally degraded images were subsequently derived for comparative purposes with Landsat 5 Thematic Mapper (TM). The baseline presence/absence maps had an overall accuracy of 67% at the Spencer study site and 85% at the Medicine Lodge study site. Unexpectedly high-accuracy results were produced from the images that were spectrally degraded to the bandwidths of Landsat 5 TM, which suggests that high spectral resolution is not critical to leafy spurge detection. However, a classification using a Landsat 5 TM image indicates that the sensor is inadequate for regional distribution monitoring. The differences in results between the actual and degraded images suggest that a sensor with comparable resolutions but improved instrumentation (e.g., signal to noise) may offer an alternative to hyperspectral data for mapping leafy spurge at regional scales.