Veterinary Toxicovigilance: Objectives,Means and Organisation in France

Veterinary Research Communications -     

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.     

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.     

P‐36 Juvenile idiopathic nasal scaling in three Bengal cats

Affected cats were three Bengals, one male and two females, whose age at onset of lesions ranged from 13‐ to 20‐weeks old. Nasal planum scaling progressed to thick crusting, consequent exfoliation and exposure of underlying erosions. No signs of pruritus or pain were present but mild respiratory signs were noticed. In all cats, haematology and biochemistry were normal, they were FIV and FeLV negative, PCR for herpesvirus, calicivirus and Chlamydia were negative, and viral isolation for calicivirus and herpesvirus was negative. Wood's lamp examination was negative, as were bacterial and fungal cultures. Cytology showed exfoliating keratinocytes. A skin biopsy taken from one case showed no significant changes. Biopsies from the nasal planum of four dead cats with nondermatological conditions (controls) were collected for comparative studies. Morphometry to record the percentage of the granular layer (GL) and stratum corneum (SC) on the total thickness of the epidermis of the nasal planum was performed. The GL and SC accounted for 10.2 and 21.7% of the epidermal thickness in the affected cat, compared to 18.3 and 20.2%, respectively, in the controls. A significant reduction (P < 0.02) of the SC thickness was detected in the affected cat compared to controls. No treatment was instituted as all cats underwent complete (two cats) or nearly complete (one cat) resolution. The reported cases share the same breed, age at onset, type of lesions and a similar outcome. A reduction of the SC thickness in one of the affected cats was recorded. Therefore, an underlying congenital condition is suspected that manifests with high epidermal cell turnover and normal keratinization. Funding: Self‐funded.     

P‐81 A retrospective study of equine sarcoidosis

The purpose of this retrospective study was to evaluate six cases of equine sarcoidosis for initial presenting symptoms, response to therapy and actual outcome. Dermatologists and dermatopathologists from Europe, the United States, Australia and Canada were contacted to obtain these six cases, as this is a rare disease. Signalment, clinical signs, histological findings, clinical management and outcome were determined via a questionnaire and compared to former reports. There was no age or breed predilection, and four of six horses were geldings. Age of onset ranged from 3 months to 17 years. Onset of the disease was insidious or rapid. Interestingly, in five of six cases, scaling began on the trunk (girth and shoulder). Scaling, crusting and alopecia were seen in all six horses. In one horse, clinical signs of systemic disease were reported and included intermittent fever, prescapular lymphadenopathy, depression, poor body condition and nasal discharge. Treatment included phenylbutazone, deworming agents, antibiotics, short‐term low‐dose corticosteroids, and 1–1.5 mg/kg of prednisolone. One horse showed a partial response to trimethoprim and sulfonamide, and five of six went into clinical remission with corticosteroid treatment. Five of six horses were still alive 1 year after diagnosis; one horse was diagnosed <12 months ago. Two horses are in complete remission 4 and 8 years after diagnosis. In both horses, clinical signs recurred after cessation of therapy and went into remission again with reintroduction of treatment. Both of these horses have been in remission for several years without therapy. Funding: Self‐funded.     

Antiviral Effect of Saccharomyces cerevisiaeβ‐glucan to Swine Influenza Virus by Increased Production of Interferon‐γ and Nitric Oxide

The aim of these experiments was to investigate the potential antiviral effect of Saccharomyces cerevisiaeβ‐glucan on the pneumonia induced by swine influenza virus (SIV). Forty colostrum‐deprived 5‐day‐old piglets were randomly divided into four groups of 10. The 20 pigs in groups 1 and 2 were administered Saccharomyces cerevisiaeβ‐glucan orally (50 mg/day/pig; En‐Bio Technology Co., Ltd) for 3 days before SIV infection and those in groups 3 and 4 were given culture medium/diluent alone. Groups 1 and 3 were inoculated intranasally with 3 ml of tissue culture fluid containing 2 × 10⁶ tissue culture infective doses 50% (TCID₅₀)/ml of SIV and those in groups 2 and 4 were exposed in the same manner to uninfected cell culture supernatant. The microscopic lung lesions induced by SIV infection (group 1 pigs) were significantly more severe than those induced by infection in animals pre‐administered β‐glucan (group 3) (P < 0.05). Significantly more SIV nucleic acid was detected in the lungs of pigs experimentally infected with SIV only (group 1) at 5, 7 and 10 days post‐inoculation (dpi) compared with lungs from pigs pre‐administered β‐glucan and infected with SIV (group 3) (P < 0.05). The concentrations of interferon‐γ (IFN‐γ) and nitric oxide (NO) in bronchoalveolar lavage fluid from pigs pre‐administered β‐glucan and infected with SIV (group 3) were significantly higher than for any other group at 7 and 10 dpi for IFN‐γ, and at 5, 7 and 10 dpi for NO (P < 0.05). Saccharomyces cerevisiaeβ‐glucan reduced the pulmonary lesion score and viral replication rate in SIV‐infected pigs. These findings support the potential application of β‐glucan as prophylactic/treatment agent in influenza virus infection.     

FC‐51 Comparison of intradermal test and antigen‐specific IgE test in 22 cases of feline allergic dermatitis

The usefulness of the intradermal test (IDT) and the serological allergy test (SAT) for detecting antigen‐specific IgE in allergic cats has not yet been established. In this study, we compared the results of IDT with those of SAT and evaluated the clinical usefulness of the two tests for detecting possible allergens in allergic cats. IDT and SAT using eight antigens were performed on 22 cats with intense pruritus after excluding ectoparasites and performing diet elimination tests. Approximately 50% of the cats reacted to at least one allergen by either IDT or SAT, and 36.4% of the cats reacted on both IDT and SAT. In contrast, seven healthy cats did not show any reactions on IDT or SAT. The most commonly detected allergen in both tests was house dust mites (IDT, 36.4%; SAT, 40.9%). Five cats reacted to one allergen and the others reacted to more than one allergen with IDT. Three cats reacted to one allergen with SAT. The following percentage agreement between the results of the two tests was calculated: house dust mites (86.4%), cat fleas (63.6%), grass mix (86.4%), common mugwort (81.8%), cat epithelia (90.9%), ragweed (86.4%), Japanese cedar (90.9%), and plantain (81.8%). The overall mean percentage agreement was 83.5%. In summary, the present study showed good agreement between IDT and SAT for cats, and SAT may be more sensitive than IDT, but less specific for detecting sensitized allergens. Funding: Self‐funded.     

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.