Patch test reactions to house dust mites in dogs with atopic dermatitis

The purpose of this study was to determine the percentage of dogs with spontaneous atopic dermatitis that show a positive patch test reaction to a commercially available 20% house dust mites mixture containing equal parts of Dermatophagoides farinae and Dermatophagoides pteronyssinus in white petrolatum. In addition, we evaluated whether skin reactions induced after the epicutaneous application of house dust mites were clinically and histologically similar to naturally developed skin lesions of dogs with atopic dermatitis. Furthermore, we investigated if the reactions induced by house dust mites were true allergic reactions by comparing them to atopic lesional skin and to patch test reactions induced by an irritant substance (sodium lauryl sulphate). White petrolatum alone and nonlesional skin sites were used as negative controls. Macroscopic and microscopic evaluations of the patch test and control sites were performed in a blinded fashion at 48 and 72 h after patch test application. Microscopic results were evaluated in a qualitative and quantitative manner. A chi‐square test for homogenicity was used for the quantitative analysis to compare the proportion of each dermal inflammatory cell type among positive histopathological tested sites. P values ≤ 0.05 were considered significant. The study included 12 healthy nonatopic dogs and 13 dogs with nonseasonal atopic dermatitis. None of the nonatopic dogs reacted to house dust mites and white petrolatum. Ten (77%) of the 13 atopic dogs reacted macroscopically and histopathologically to house dust mites. Macroscopic reactions induced by house dust mites were characterized by erythema, oedema and papules. The macroscopic reactions induced by house dust mites were identical to lesional skin in 20% of the dogs and identical to reactions induced by sodium lauryl sulphate in 40% of the dogs. Qualitative histopathological findings showed that the reactions induced by house dust mites were similar to atopic lesional skin in 80% of the dogs and were similar to sodium lauryl sulphate in 20% of the dogs. Quantitative analyses showed that the proportion of neutrophils in reactions induced by sodium lauryl sulphate was significantly higher (P < 0.05) compared to house dust mites reactions, which could be a differentiator factor between an allergic and an irritant reaction. These results showed that the epicutaneous application of house dust mites in dogs with atopic dermatitis induced histopathological lesions similar to spontaneous atopic lesions in dogs. Therefore, this study demonstrated that house dust mites penetrated the skin of dogs with atopic dermatitis and induced an inflammatory response that resembled a true allergic reaction. Funding: Small Companion Animal Grant, University of Minnesota.     

Fortification of Wheat Dough with Calcium and Magnesium Ions Affects Empirical Rheological Properties

This study evaluated the influence of calcium and magnesium ions on the empirical rheological properties of wheat flour to verify possible effects of these ions on processing because, in addition to their nutritional importance, they are also responsible for water hardness. Calcium (0–1.30 g/100 g) and magnesium (0–0.34 g/100 g) ions from sulfate salts were added to wheat flour, according to a central composite rotatable design. The farinograph and extensigraph properties of wheat flour and its mixtures were evaluated. The results were analyzed by response surface methodology. Calcium ions stood out for increasing water absorption, decreasing mixing stability, and producing a delayed effect on dough extensibility (reduced at 135 min). Magnesium ions influenced most flour rheological properties in a similar manner to oxidizing agents (increased dough stability, increased resistance to extension, and reduced extensibility), thus proving to be a possible replacement agent for these additives. An interaction effect of the combined calcium and magnesium ions was observed on dough development time. The results showed that effects on processing can occur when wheat flour fortification is made, and adaptations on wheat flour specifications, product formulation, and processing parameters may be required.     

Characterization of symptoms of senescence and chilling injury on inflorescences of Heliconia bihai (L.) cv. Lobster Claw and cv. Halloween

Inadequate temperatures during the shipping and commercialization of cut tropical flowers may accelerate the senescence process and cause chilling injury, leading to symptoms that have not yet been described for Heliconia bihai. The aim of the present study was to evaluate physiological responses in cut inflorescences of H. bihai cv. Lobster Claw (LC) and cv. Halloween (HW) as well as symptoms of senescence and chilling injury. For such, changes in fresh weight, bract color (L*, a* and b*), percentage of absolute integrity (PAI) of cell membranes and leakage of potassium ions (LPI) were determined. The flowering stems were evaluated at five different intervals after harvest (0, 2, 4, 6 and 8 d). A refrigerated treatment (RT) with a temperature of 6.5 °C and 85% relative humidity was compared to a control treatment (CT) at room temperature of 24 °C and 66% relative humidity. Both cultivars stored at 6.5 °C exhibited dryness of bract tissue (symptom of senescence) and dark stains that became brownish and evolved to necrosis (symptom of chilling injury). The visual quality of inflorescences decreased with time in both cultivars maintained without refrigeration. The severity of chilling injury increased with the length of storage time in both cultivars. There was a significant reduction in the fresh weight of inflorescences in both treatments (RT and CT) and both cultivars (LC and HW). Bract color changed in both cultivars at 6.5 °C. There was no change in PAI throughout the evaluation period in the inflorescences stored at room temperature, whereas those stored at 6.5 °C for 6 and 8 d had lower PAI values. The inflorescences in the control treatment underwent no change in LPI values, whereas those stored under refrigeration had increased LPI values after the sixth day of storage. The physiological responses of cut Heliconia flowers were influenced by storage period and temperature, as demonstrated by visual symptoms of chilling injury and senescence.     

Short and long‐term distribution with depth of soil organic carbon and nutrients under traditional and conservation tillage in a Mediterranean environment (southwest Spain)

The impacts of tillage and cropping sequences on soil organic matter and nutrients have been frequently reported to affect the uppermost soil layers, but there is little published information concerning effects at greater depth. This article reports results on the distribution of soil organic carbon (SOC), active carbon (AC), N, Olsen‐P and extractable K within 100 cm in short (4 yr) and long (16 yr) term experiments under different tillage systems. Short (TT₄) and long (TT₁₆) traditional tillage are compared with conservation tillage, reduced (RT₁₆) and non‐tillage (NT₄). The results show more accumulation of SOC in the near‐surface under RT₁₆ and NT₄ in both experiments compared with traditional tillage. Moreover, greater C content occurs to 40 cm depth in the long‐term experiment. The results demonstrate the importance of time on C accumulation, not only in near‐surface layers but also at greater depths. Active C is an indicator of the increase in soil quality in the long‐term experiment. This trend is only apparent for the first 10 cm in the short‐term experiment. Patterns in N, Olsen‐P and extractable K are similar to that of SOC. However, only extractable K is significantly greater in soil under conservation tillage (RT₁₆ and NT₄) after short and long periods. Potassium availability is a good indicator of the changes caused by tillage. Our results indicate that studies of soils at depth could be very useful in long‐term experiments to demonstrate the effect of conservation tillage on C and nutrient distribution.     

Extrinsic and intrinsic factors influence daily activity of a Mediterranean cyprinid

Factors shaping individual behavior remain poorly understood for European cyprinids. We examined the effects of extrinsic (temperature and year) and intrinsic factors (body size and shoal size) on the daily activity of chub Squalius torgalensis, a critically endangered cyprinid found in the Torgal stream, Portugal. We quantified chub behaviors while snorkeling, using instantaneous sampling of focal individuals, in spring 2009 and 2010. We used Principal Component Analysis to describe individual behavior and assessed competing models relating individual behavior to explanatory variables using Akaike's Information Criterion. Year had a strong influence on chub behavior, with individuals spending more time drift foraging and orienting in 2009, whereas in 2010 they spent more time benthic foraging and cruising. Shoal size did not influence behaviors in 2009, but in 2010 individuals in small shoals foraged benthically and searched with greater frequency than those in large shoals. Individuals increased cruising and decreased searching as they grew in length, but the opposite trend was observed as temperature increased. Our results indicate that both extrinsic and intrinsic factors influence daily activity of chub, with individual behaviors varying between years, and depending upon shoal size, body size and temperature. This suggests that chub display high levels of individual behavioral flexibility, which may be important for fish inhabiting harsh and highly variable Mediterranean streams. Conservation efforts for this species will benefit by integrating behavioral information into management strategies.     

Disseminated pythiosis in three horses

Three cases of equine subcutaneous pythiosis with dissemination to the internal organs were investigated. The subcutaneous lesions were observed on the mammary gland, nostrils and limbs of the infected horses. Histopathological analysis of the infected tissues revealed a strong eosinophilic reaction, with macrophages, mast cells and giant cells. Sparsely septated hyphal filaments of 4-6 microm diameter were identified in the center of the eosinophilic areas. Specific fluorescent antibody against Pythium insidiosum confirmed the hyphae in the infected tissues in three examined horses. In one of the three cases, the DNA sequences amplified from the infected subcutaneous tissues and internal organs, revealed that P. insidiosum's 18S SSU rDNA amplicons shared 100% identity with those sequences deposit in GenBank. This is the first report confirming by immunochemical and genetic techniques that P. insidiosum can disseminated from superficial to deep structures.     

Evaluation of a new immunocapture test for the diagnosis of ovine brucellosis caused by Brucella melitensis

A new immunocapture technique has been applied to the diagnosis of ovine brucellosis under experimental conditions. The tests were made on a serum bank derived from both young and adult ewes vaccinated conjunctivally with the Rev 1 strain at a dose of 10(8) to 10(9) colony-forming units. Adult ewes were infected experimentally two-and-a-half years after they had been vaccinated and the results were compared with an unvaccinated control group. The condition of each animal in terms of infection with Brucella melitensis was determined by clinical and bacteriological investigations. The development of the immune response was compared by the rose bengal test, the complement fixation test, the Coombs' test and the immunocapture technique for 180 days after the vaccination and for 410 days after the experimental infection, that is, the two following gestations. The results suggest that the new technique is more specific in animals vaccinated conjunctivally, regardless of their age when they were vaccinated. After the experimental infection, significantly (P < 0.05) fewer of the vaccinated sheep which were free of clinical signs and were not excreting B melitensis reacted positively to the test.     

Assessment of relevant factors and relationships concerning human dermal exposure to pesticides in greenhouse applications

Principal component analysis (PCA) was applied to the gas chromatographic data obtained from 23 different greenhouse trials. This was used to establish which factors, including application technique (very small, small, medium and large drop-size), crop characteristics (short/tall, thin/dense) and pattern application of the operator (walking towards or away from the treated area) are relevant to the dermal exposure levels of greenhouse applicators. The results showed that the highest exposure by pesticides during field applications in greenhouses, in the climatic conditions and in the crop conditions typical of a southern European country, occurs on the lower legs and front thighs of the applicators. Similar results were obtained by hierarchical cluster analysis (HCA). Drop-size seems to be very important in determining total exposure, while height and density of crops have little influence on total exposure under the conditions of the present study. No pesticide type is a major factor in total exposure. The application of multiple regression analysis (MRA) allowed assessment of the relationships between the pesticide exposure of the less affected parts of the body with the most affected parts.