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.     

FC‐61 The role of a liquorice preparation in healing of experimental wounds in calves: a histopathologic study

Liquorice, a derivative of the root of Glycyrrhizia glabra, has been proposed as a treatment for gastric ulcers. Here, a preparation of liquorice combined with sesame oil was evaluated in experimental wound healing. The study was carried out in two female 10‐month‐old Holstein calves. An area of 10 × 10 cm on each side of the thorax of the animals was surgically prepared and infiltrated with local anaesthetic. Eight uniform skin wounds were created in two rows with an 8‐mm biopsy punch on each prepared area. Each side of an animal was used to evaluate one treatment (Groups 1–4). To make the mixture, powdered and sieved liquorice (16 g) was mixed with sesame oil (36 g) to make a suspension. The wounds were treated for 7 days. The wounds of Groups 1–3 were rinsed daily with normal saline, after which the mixture of liquorice and sesame oil (Group 1) or only the sesame oil (Group 2) was applied to the wounds. In Group 4, no treatment was applied. All wounds were left open. On day 8, skin biopsies were taken from the wounds for histopathologic study. In Group 1, seven wounds (87%) showed moderate granulation tissue, mild scab formation, and complete re‐epithelialization. In Group 2, there was massive scab formation with no granulation tissue and minimal re‐epithelialization. In Groups 3 and 4, mild granulation tissue, severe scab formation and very limited re‐epithelialization were observed. We conclude that a preparation of liquorice and sesame oil can be an effective remedy for wound healing. Funding: Self‐funded.     

Epidemiological Study of Non‐contagious Intramammary Infections in Nine Commercial Dairy Herds following a Staphylococcus aureus Control Programme

Changes in prevalence in intramammary infection, by pathogen type, in herds applying a stringent contagious mastitis control programme was studied. Enrollment of 1651 lactating cows and collection of milk samples was made in this ancillary study to a cohort study of the dynamics of mastitis prevalence after adoption of a strict contagious mastitis control programme that targeted the elimination of mastitis caused by Staphylococcus aureus. Nine commercial dairies in Italy were used. Aseptic collection of milk samples from all lactating cows was performed at the time of enrollment, from all cows within 7–14 days of entering the lactating herd after the date of enrollment, and from all lactating cows at 2, 4, 7, 10 and 14 months after the date of enrollment. Prevalence of intramammary infection by pathogen type was determined from culture of milk samples. Application of the strict contagious mastitis programme did not lead to an increased risk of non‐contagious mastitis. The risk of coliform, environmental streptococcal and coagulase‐negative staphylococcal intramammary infections decreased after adoption of the programme. The data reported herein indicate that the overall risk for any intramammary infections decreases with adoption of a strict contagious mastitis programme, and that such a programme therefore does not necessarily lead to an increase in environmental mastitis.     

The effect of sowing date, stale seedbed, row width and mechanical weed control on weeds and yields of organic winter wheat

Three field experiments were carried out in organically grown winter wheat in Denmark. The treatments were sowing time (normal or late sowing) and false seedbed, row width (12 and 24 cm) and weed control method [untreated; mechanical weed control (weed harrowing at 12 cm supplemented with inter‐row hoeing at 24 cm); and herbicide weed control]. Weed biomass in midsummer was greatest on plots sown at the normal sowing time (compared with delayed sowing) and was reduced by mechanical or chemical weed control (compared with untreated plots). Row width alone had no influence on weed biomass, but in the experiment with high weed pressure, the more intensive mechanical weed control used at a row width of 24 cm reduced weed biomass. Normal sowing time tended to give higher yields, but this was only statistically significant in one of the three experiments. Wide rows gave a yield decrease in the experiment with low weed pressure. The effect of weed control on yield was dependent on the weed pressure. At low weed pressure, mechanical weed control caused a yield decrease compared with untreated or herbicide treated. At intermediate weed levels there were no differences, whereas at high weed pressure, mechanical weed control and herbicide treatment caused a yield increase compared with untreated. False seedbeds were shown to contribute to a decrease in the soil seed reserve.     

Yeasts as antagonists against fireblight

Yeasts are potential antagonists of microorganisms in the phyllosphere. Due to their osmotolerance, they should also be able to colonize apple flowers. In field experiments, we applied yeast agents against fireblight at two different sites in the southern part of Germany. They showed efficiencies 0–20% below Plantomycin (streptomycin). Co-culture experiments in liquid basal media with synthetic nectar resulted in suppression of Erwinia amylovora by yeast. This effect could not be confirmed with population studies of yeasts and E. amylovora in flower clusters. Field and laboratory experiments indicated that yeasts have antagonistic properties against fireblight but further research is needed to investigate this potential.     

Détection moléculaire spécifique de la région vir du plasmide pTi d'Agrobacterium tumefaciens dans les sols et plants au Maroc

Le meilleur moyen de contrôle du crown gall ( Agrobacterium tumefaciens ) est la prévention et donc la détection de la bactérie pathogène dans les sols prévus pour les pépinières. Pour cela nous avons essayé une approche moléculaire utilisant la PCR pour caractériser le plasmide pTi par le couple des amorces spécifiques F14/F44 dans les sols de diverses régions marocaines. Cette technique de détection représente un indicateur potentiel d'avertissement sur l'état d'infection des sols et plants par A. tumefaciens qui permet ainsi de déclencher la lutte biologique par le NoGall, produit commercial australien, hébergeant l'antagoniste K₁₀₂₆.     

Non-target toxicology of a new mosquito larvicide, trypsin modulating oostatic factor

Trypsin modulating oostatic factor (TMOF), a peptide hormone originally isolated from the ovaries of adult Aedes aegypti, is currently under commercial development as a new pesticide chemistry with a novel mode of action for the control of larval mosquitoes. The objective of the current research is to evaluate potential risks of the use of TMOF as an insecticide on non-target organisms. TMOF (YDPAP₆) was degraded in vitro (as determined by HPLC and LC/MS) to DPAP₆, PAP₆, and then AP₆ by leucine aminopeptidase, a pancreatic enzyme found in the digestive system of vertebrates. The rate of degradation of TMOF and PAP₆ was significantly greater than that of DPAP₆, while no metabolism of AP₆ was found. TMOF technical insecticide was produced on a commercial scale by recombinant yeast (heat-killed before application). The technical TMOF when administered in a single dose by gavage to male and female mice at 2000 mg dry weight/kg body weight produced no negative effects as compared to controls up to 12 days after treatment. When male and female mallard ducks were treated by gavage with 1250 mg dry weight of technical TMOF/kg body weight each day for 5 days, again no toxic effects were noted through 35 days after the last treatment. TMOF technical insecticide was also applied to the shaved skin of male and female rabbits at the rate of 2000 mg/kg for 1-2 days, with no effect. The end point observations in these in vivo experiments were mortality; changes in growth rate, behavior, body structure, and color; and possible lesions observed during necropsy. Finally, Daphnia incubated with technical TMOF in rearing water at the level of 1.0 × 10⁶ yeast cells/ml (10 mg/ml) also demonstrated no negative effects on mortality, growth, molting, time to first brood, and production of viable neonates. It appears from these studies that TMOF can be degraded by vertebrate digestive proteases and technical TMOF is not toxic to the non-target organisms examined.     

Interfertility of Two Mating Populations in the Gibberella Fujikuroi Species Complex

Gibberella fujikuroi and Gibberella intermedia(mating populations ‘C’ and ‘D’ of the G. fujikuroi species complex) can be distinguished by differences in the spectrum of mycotoxins produced, the lack of sexual cross-fertility and diagnostic differences in their DNA sequences. Some isolates from these two biological species, however, can interbreed and complete meiosis to produce viable progeny. Analysis of marker segregation amongst such hybrid progeny can be used to estimate the degree of genomic rearrangement and genetic incompatibility that has accumulated since these sibling species diverged. Recombinant progeny were isolated from crosses of the standard tester strains for these two species and from crosses between these standard testers and a field isolate (KSU X-10626) that was cross-fertile with tester strains of both species. Progeny in all of the crosses segregated for amplified fragment length polymorphisms (AFLPs). Segregation of AFLP loci deviated from 1:1 for two thirds of the loci amongst the progeny of the cross between the ‘C’ and ‘D’ mating population tester strains, but <20% of the polymorphic loci in the cross of either tester with KSU X-10626 showed such distortion. It was concluded that G. intermedia and G. fujikuroi are sufficiently interfertile to belong to the same biological species, but that changing the nomenclature to reflect this interfertility requires more evidence for the natural occurrence of a continuum in fertility than is presently available.     

PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus

Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination of grapes and their by-products by ochratoxin A. A PCR-based method was developed to detect DNA of A. carbonarius and A. japonicus. Two pairs of primers (CARBO1/2 and JAPO1/2) designed from the calmodulin gene, produced PCR products of 371 and 583 bp for A. carbonarius and A. japonicus, respectively. Primer specificity was tested with DNA of 107 strains belonging to Aspergillus section Nigri isolated mostly from grapes in Europe. The sensitivity of primers CARBO1/2 and JAPO1/2 was 12.5 pg when using pure total genomic DNA of the two species. The developed primers provide a powerful tool for detection of the main ochratoxigenic producing Aspergillus species in grapes.     

Effectiveness of plant extracts of Paeonia suffruticosa and Hedera helix against diseases caused by Phytophthora infestans in tomato and Pseudoperonospora cubensis in cucumber

Journal of Plant Diseases and Protection - The effects of ethanolic extracts (20 % per fresh weight (Fw) or 6 % per dry weight) of 20 different plant species were tested against late blight...