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.     

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.     

A perspective on the measurement of time in plant disease epidemiology

The growth and development of plant pathogens and their hosts generally respond strongly to the temperature of their environment. However, most studies of plant pathology record pathogen/host measurements against physical time (e.g. hours or days) rather than thermal time (e.g. degree-days or degree-hours). This confounds the comparison of epidemiological measurements across experiments and limits the value of the scientific literature.     

Effects of straw and silicon soil amendments on some foliar and stem-base diseases in pot-grown winter wheat

Four foliar and two stem-base pathogens were inoculated onto wheat plants grown in different substrates in pot experiments. Soils from four different UK locations were each treated in three ways: (i) straw incorporated in the field at 10 t ha⁻¹ several months previously; (ii) silicon fertilization at 100 mg L⁻¹ during the experiment; and (iii) no amendments. A sand and vermiculite mix was used with and without silicon amendment. The silicon treatment increased plant silica concentrations in all experiments, but incorporating straw was not associated with raised plant silica concentrations. Blumeria graminis and Puccinia recondita were inoculated by shaking infected plants over the test plants, followed by suitable humid periods. The silicon treatment reduced powdery mildew ( B. graminis ) substantially in sand and vermiculite and in two of the soils, but there were no effects on the slight infection by brown rust ( P. recondita ). Phaeosphaeria nodorum and Mycosphaerella graminicola were inoculated as conidial suspensions. Leaf spot caused by P. nodorum was reduced in silicon-amended sand and vermiculite; soil was not tested. Symptoms of septoria leaf blotch caused by M. graminicola were reduced by silicon amendment in a severely infected sand and vermiculite experiment but not in soil or a slightly infected sand and vermiculite experiment. Oculimacula yallundae (eyespot) and Fusarium culmorum (brown foot rot) were inoculated as agar plugs on the stem base. Severity of O. yallundae was reduced by silicon amendment of two of the soils but not sand and vermiculite; brown foot rot symptoms caused by F. culmorum were unaffected by silicon amendment. The straw treatment reduced severity of powdery mildew but did not detectably affect the other pathogens. Both straw and silicon treatments appeared to increase plant resistance to all diseases only under high disease pressure.     

Fireblight monitoring in Lithuania1

Lithuania has requested that its whole territory should be recognized by the EU as a protected zone for Erwinia amylovora . Fireblight monitoring was performed in 1998/2002 with the aim of detecting and identifying the bacterium, and of determining its distribution in the country. The study consisted of periodic surveys (at least twice a year) of nurseries, orchards, collective farms and host plants, growing individually or in small groups, as well as the surrounding zone within a radius of 250 m. Tests, under conditions of quality control, were applied to host plants with and without symptoms, using detection methods such as ELISA and immunofluorescence (with polyclonal antibodies), semi-selective plating and pathogenicity.     

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.     

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.     

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.     

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.