Identification of Yersinia enterocolitica in Minced Meat: A Comparative Analysis of API 20E,Yersinia Identification Kit and a 16S rRNA‐based PCR Method

The isolation and identification of Yersinia enterocolitica from minced meat on CIN agar medium is still one of the major problems in food microbiology because of the low selectivity of cefsulodin–irgasan–novobiocin (CIN) agar. A total of 198 minced meat samples were collected from commercial establishments (butcher shops and supermarkets) in seven German cities in order to investigate the sensitivity and specificity of three identification techniques suitable for the differentiation of Y. enterocolitica within the rich background flora on CIN agar plates. As expected isolation of Y. enterocolitica from minced meat on CIN agar medium after 72 h enrichment in peptone, sorbitol and bile salts (PSB) broth was difficult because all plates were abundantly covered with numerous ‘typical’Yersinia‐like colonies of bull's eye appearance as well as with atypical colonies. Based on the phenotype of the colonies it was possible to detect colonies showing Yersinia‐like growth on CIN agar in 52 samples (26%). For identification of Y. enterocolitica the API 20E system (bioMerieux, Nürtingen, Germany), the Yersinia identification kit (Merlin, Bornheim‐Hersel, Germany) and a 16S rRNA based PCR assay were compared. Only in one sample (0.5%) a Y. enterocolitica strain was detected by all methods. Of the three identification systems tested for routine laboratory diagnostics the API 20E system was found to be the most suitable tool to identify Y. enterocolitica colonies within the rich background flora from minced meat samples on CIN agar plates.     

Effect of recombinant canine IFNγ on canine atopic dermatitis evaluated by clinical signs,histopathology and expression of Th1/Th2 cytokine mRNA

Atopic dermatitis (AD) is thought to be caused by immunologic abnormalities expressed as a Th1/Th2 cytokine imbalance in both humans and dogs. Several studies have focused on the therapeutic effects of IFNγ in human AD with successful results; however, the mechanism of action of IFNγ is not fully understood. We investigated the effect of recombinant canine interferon gamma (rCaIFNγ) on 10 dogs with AD and evaluated the ratio of IL‐4 mRNA to IFNγ mRNA in peripheral blood mononuclear cells, serum total IgE levels, and histological changes in skin. After six injections of rCaIFNγ over a span of 2 weeks, seven of the 10 dogs showed improvement, and six of these seven dogs exhibited decreased IL‐4:IFNγ mRNA ratios. Two of the three cases that did not improve had increased IL‐4:IFNγ mRNA ratios. Total serum IgE levels were significantly decreased in nine of 10 cases. The number of IgE‐positive cells detected by immunostaining and the number of mast cells in skin biopsy samples were decreased. A reduction of epidermal cell layers was demonstrated by histopathology after treatment. These results demonstrated that rCaIFNγ may be a novel safe and effective therapeutic option for the treatment of canine AD, and the mechanism of action of rCaIFNγ may be related to the modulation of Th2 cytokines to Th1 cytokines with the reduction of serum IgE production. Funding: Self‐funded.     

Significance of β2‐Toxigenic Clostridium perfringens Infections in Animals and Their Predisposing Factors – A Review

The novel β2‐toxin of Clostridium perfringens has recently been described as the cause of enteric diseases in animals. The biological activity of β2‐toxin is similar to that of the β1‐toxin with a possibly weaker cytotoxic activity. However, the production of β2‐toxin in vitro is not seen in all β2‐toxin‐gene (cpb2)‐positive C. perfringens strains, and to deduce a clinical importance solely from the detection of cpb2 is difficult. Detection of cpb2‐positive C. perfringens from various animal species with and without enteric diseases demonstrates the wide distribution of cpb2 in nature, and the presence of cpb2 gene is therefore not considered a risk by itself. Predisposing factors like low trypsin activity in the intestinal tract, antibiotic and/or antiphlogistic treatment or changes in diet can result in the selection of β2‐toxigenic C. perfringens which may lead to enteritis or enterotoxaemia.     

The effect of non-inversion tillage and light availability on dispersal and spatial growth of Cynodon dactylon

The invasive ability of Cynodon dactylon is dependent on self dispersal and on cultivation practices. Tillage can seriously change patch biomass and spatial structure, spreading vegetative propagules of the weed. The objectives of this study were: (i) to quantify the effect on non‐inversion tillage on dispersal, establishment and colonization of C. dactylon and (ii) to propose a simple model considering soil cultivation effects and light availability on spatial growth of weed patches. Two experiments were carried out, exploring different soils and environmental conditions. Spatial distribution of vegetative units differed when tillage was conducted with different non‐inversion implements and could be described by simple functions. A minimum patch biomass seems necessary before vegetative structures are vulnerable to movement by cultivation. Only a small proportion of the biomass dispersed from original patches was able to establish. However, simulation showed that the area colonized by C. dactylon mostly increased by means of tillage dispersal, both with and without crop competition, in one growing cycle. It appears sensible to consider changing cultivation practices to reduce weed dispersal and to use crop competition for light to create unsuitable habitats limiting weed colonization.     

UV-absorbing compounds and susceptibility of weedy species to UV-B radiation

Stratospheric ozone (O₃) depletion has led to increased terrestrial ultraviolet‐B (UV‐B) radiation (290–320 nm). Leaves exposed to this radiation produce UV‐absorbing compounds in the epidermal cells, which protect plants from UV‐B damage. To determine the role of UV‐absorbing compounds in the UV‐B sensitivity of weeds (common chickweed (Stellaria media), downy brome (Bromus tectorum), green smartweed (Polygonum scabrum), redroot pigweed (Amaranthus retroflexus), spotted cat’s‐ear (Hypochoeris radicata), and stork’s‐bill (Erodium cicutarium)) seedlings were exposed to 0, 4 (field ambient), 7 (18% O₃ depletion) and 11 (37% O₃ depletion) kJ m^?2 d^?1 of biologically effective UV‐B radiation in a greenhouse. Ultraviolet‐absorbing compounds were extracted from the second true‐leaf (0.5 cm² samples) with methanol : distilled water : HCl (79 : 20 : 1) in an 85°C water bath for 15 min, and the absorbance of the extracts measured at 300 nm. The shoot dry biomass was recorded to determine the susceptibility to UV‐B radiation. Common chickweed was the most sensitive and green smartweed the least sensitive weed to UV‐B radiation. The latter accumulated more UV‐absorbing compounds and this accumulation occurred earlier compared with common chickweed. As UV‐B_BE radiation levels increased from 0 to 11 kJ m^?2 d^?1, the green smartweed shoot biomass did not decline. However, the biomass of all five susceptible species declined despite an increase in the UV‐absorbing compounds in response to increased UV‐B radiation. Therefore, formation of a ‘UV‐screen’ in these species is not sufficient to fully prevent UV‐B damage. When the concentration of UV‐absorbing compounds in the six species was plotted against their susceptibility to UV‐B radiation, no relationship was observed. Thus, while the accumulation of UV‐absorbing compounds may be a major factor in the protection of certain species against UV‐B radiation and may offer some degree of defence in other species, it does not explain UV‐B susceptibility differences in weedy species in general.     

Analysis of a Secreted Aspartic Peptidase Disruption Mutant of Glomerella cingulata

Peptidases have been implicated in the pathogenicity of fungi that cause disease in plants. Expression of the secreted aspartic peptidase gene (gcsap), of a Glomerella cingulata isolate pathogenic on apples, is induced during appressorium formation. To determine whether the secreted aspartic peptidase (GcSAP) is essential to pathogenicity, gcsap was disrupted using a vector containing a 637 bp fragment of genomic DNA that encodes the sequence spanning the two active site aspartic acid (Asp) residues. To ensure that the truncated gcsap gene products could not have residual peptidase activity the codons for the active site residues Asp¹¹² and Asp²⁹⁷ were both mutated to histidine residues. Both PCR and Southern analysis confirmed disruption of gcsap. Neither gcsap mRNA nor GcSAP activity was detected in the disruption mutant. Pathogenicity tests on fruit from three apple cultivars showed that GcSAP was not required for pathogenicity. The disruption mutant grew on medium containing protein as the sole source of nitrogen because G. cingulata secretes a previously undetected peptidase(s). A serine peptidase that had a pH optimum between pH 7.0 and 8.0 and a K _m of 0.25 mM for the synthetic substrate succinyl-Ala–Ala–Pro–Phe-p-nitroanilide was identified.     

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.     

Differences in the growth stages of Erysiphe pisi on cultivars of field pea (Pisum sativum L.)

Journal of Plant Diseases and Protection - The infection process of Erysiphe pisi on field pea cvs. ‘Highlight’ and ‘Tara’ carrying resistance gene erl, line...     

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...