S100A4 (metastasin) positive mesenchymal canine mammary tumour spheroids reduce Tenascin C synthesis under DMSO exposure in vitro

In breast cancer research S100A4‐positive tumour‐associated stromal cells are assumed as primary source of Tenascin C (TNC) in the metastatic environment. Aim of the present study was to isolate and characterize S100A4/TNC positive stromal canine mammary tumour (CMT) cells. Cells grown as scaffold‐free spheroids were investigated for S100A4, TNC, and proliferative activity under 1.8% DMSO stimulation by means of Western blot and immunohistochemistry. DMSO is a commonly used drug solvent despite well‐known side effects on cells including TNC expression. DMSO did not affect proliferation, but TNC was significantly reduced under DMSO exposure for 7 and 14 days, whereby for S100A4 a reducing effect was only observed after 14 days. Without DMSO, cells stably expressed TNC and S100A4 which makes them suitable to be used in experimental approaches requiring S100A4/TNC expressing CMT stromal cells. Results show that 1.8% DMSO should not be used as solvent for experiments concerning TNC/S100A4 expression.     

Oral dosage regimen in the nonlinear pharmacokinetics of sulphadimethoxine in pigs

Summary

An oral high dosage regimen of sulphadimethoxine (SDM) was examined in pigs. The dose (50 mg/kg) in the therapeutic range, showed nonlinear pharmacokinetics, and administered by drench once a day for 4 days. The unbound plasma concentration‐time profile was compared with that of the dosage regimen based on nonlinear pharmacokinetics, where a pharmacokinetic model and parameters were used except for the first order absorption rate constant (ka) and bioavailability (F). F and ka were obtained from oral and intravenous administration of 20 and 10 mg/kg of SDM. The unbound plasma concentration was observed almost within the setting range by the dosage regimen through the experimental period. This result suggested that the dosage regimen, based on the nonlinear pharmacokinetic model, resulted in an appropriate effect in the clinical use.     

An investigation into the feasibility of interspinous injections in the horse

Injections of substances into the interspinous spaces between the thoracic and lumbar vertebrae cannot be practically achieved in the horse.     

Two cases of dermatophytic pseudomycetoma in the dog: an immunohistochemical study

Two cases of canine dermatophytic pseudomycetoma resulting in subcutaneous nodules resembling those previously reported in Persian cats are described. Culture performed from one nodule yielded dark yellow colonies consistent with Microsporum canis. Immunohistochemistry, using rabbit anti-M. canis, demonstrated specific binding to fungal elements in paraffin sections. The specificity of the antiserum was further tested by an agar gel immunodiffusion assay using a soluble extract from a feline isolate of M. canis as antigen. The antiserum did not cross-react with an Aspergillus fumigatus antigen. These are the first two reported cases of canine dermatophytic pseudomycetoma and immunohistochemical staining supported the diagnosis.     

An analysis of factors underlying hypotrichosis and alopecia in Irish Water Spaniels in the United Kingdom

A survey on the occurrence of dermatoses in the Irish Water spaniel (IWS) was carried out in the United Kingdom. A group of 20 dogs was selected and examined clinically. All dogs had a nonpruritic, noninflammatory, regionalized hair loss affecting the same areas of the body in males and females, although an initial cyclical pattern associated with the oestrus cycle was identified. Hormonal investigations showed features suggestive of an abnormality of steroidogenesis. Histopathology revealed features similar to canine recurrent flank alopecia (CRFA) and follicular dysplasia associated with abnormal melanization, as in colour dilution alopecia, although the clinical features did not correlate with those conditions. Dietary changes improved coat and skin quality in most of the cases in this series but the role of the diet was not investigated further. This study suggests that hair loss in IWS is influenced by dietary factors and sexual hormones. Abnormalities of the steroidogenic pathways may contribute to the severity of the condition.     

Verification of the fCAL turbo immunoturbidimetric assay for measurement of the fecal calprotectin concentration in dogs and cats

The concentration of calprotectin in feces (fCal) is a clinically useful marker of chronic gastrointestinal inflammation in humans and dogs. No commercial assay is widely available to measure fCal in small animal medicine, to date. Thus, we verified the immunoturbidimetric fCAL turbo assay (Bühlmann) of fCal for canine and feline fecal extracts by determining linearity, spiking and recovery, and intra-assay and inter-assay variability. We determined RIs, temporal variation over 3 mo, and effect of vaccination and NSAID treatment. Observed:expected (O:E) ratios ($\bar{x}$ ± SD) for serial dilutions of feces were 89–131% (106 ± 9%) in dogs and 77–122% (100 ± 12%) in cats. For spiking and recovery, the O:E ratios were 90–118% (102 ± 11%) in dogs and 83–235% (129 ± 42%) in cats. Intra- and inter-assay CVs for canine samples were ≤19% and ≤7%, and for feline samples ≤22% and ≤21%. Single-sample RIs were <41 μg/g for dogs and <64 μg/g for cats. With low reciprocal individuality indices, using population-based fCal RIs is appropriate, and moderate fCal changes between measurements (dogs 44.0%; cats: 43.2%) are considered relevant. Cats had significant (but unlikely relevant) fCal increases post-vaccination. Despite individual fCal spikes, no differences were seen during NSAID treatment. The fCAL turbidimetric assay is linear, precise, reproducible, and sufficiently accurate for measuring fCal in dogs and cats. Careful interpretation of fCal concentrations is warranted in both species during the peri-vaccination period and for some patients receiving NSAID treatment.     

Immunohistochemical detection of virus antigen in the nasal planum of rabid dogs

The rabies virus is one of the most neurotropic of all viruses infecting mammals. During the terminal phases of infection, the virus spreads to peripheral tissues, including the skin. The external skin of the nose, called the nasal planum, is a sensory organ where numerous nerve bundles and terminal nerves are distributed. Therefore, the nasal planum is expected to serve as a postmortem diagnostic material. However, the distribution of rabies virus antigens in the nasal planum in rabid animals has not yet been studied. In this study, the nasal planum was obtained from 45 rabid dogs. In all rabid dogs, the viral antigen was detected in the peripheral nerve tissues, Merkel cells, and squamous cells. The viral antigen in the epidermis exhibited three patterns: first, a diffuse positive pattern from the basal layer to the squamous layer; second, a reticular positive pattern along the cell membrane in the squamous layer; and third, a basal layer pattern of the epidermis. In the dermis, viral antigens were detected more often in lamellated corpuscles just beneath the rete pegs. These results suggest that the nasal planum could serve as a useful alternative source for postmortem diagnosis in rabies endemic countries.     

Unilateral malignant melanoma of the adrenal gland with metastatic spread to the thorax in a horse

A 7-year-old grey horse that had a history of inappetence, weight loss, weakness, dyspnoea, coughing and intermittent colic was examined post-mortem. A melanoma, confirmed histologically, was found in the right adrenal gland and had eroded into the posterior vena cava. Thoracic metastases were also identified. The skin, including the legs, trunk and perineum was normal with no evidence of melanomas. This was considered to be a primary, malignant melanoma, a novel finding in the adrenal gland of the horse.     

Sequelae and complications of Streptococcus equi subspecies equi infections in the horse

Streptococcus equi ssp. equi infection in the horse, or strangles, commonly results in abscessation of the submandibular, submaxillary or retropharyngeal lymph nodes. Although this classical presentation of strangles is associated with a low mortality rate, complications and sequlae may worsen the prognosis and increase mortality rates. This article reviews sequelae and complictions of S. equi ssp. equi infection in the horse, including guttural pouch empyema, bastard strangles and immune mediated diseases such as purpura haemorrhagica, myopathies and myocarditis.     

Double-blind pilot study on the effects of ketoconazole on intradermal skin test and leukotriene C4 concentration in the skin of atopic dogs

Abstract The effects of ketoconazole on intradermal skin test results and on leukotriene C₄ (LTC₄) concentration in the skin of atopic dogs were evaluated in a pilot study. Twelve atopic dogs without a detectable Malassezia dermatitis were selected. All dogs had positive immedíate reaction to intradermal injection of house dust mite (HDM) at 25 PNU mL^-1. Six dogs received a control sugar tablet and six dogs received ketoconazole at 5 mg kg^-1 PO b.i.d. for 3 weeks. On days 0 and 21, intradermal injections of saline, lipopolysaccharide (LPS) and house dust mite (HDM) were performed and biopsies of the injection sites were taken at 90 min postinjection to measure the concentration of LTC₄ in the skin. Intradermal skin test results were not affected by ketoconazole therapy. Ketoconazole significantly decreased the concentration of LTC₄ that could be elicited by the intradermal injection of saline and LPS. Ketoconazole also decreased the HDM-induced LTC₄ but differences between the prevalues and postvalues were not statistically significant. The mean decrease of LTC₄ concentration in the ketoconazole group was 37% for the saline injection, 42% for the LPS injection and 26% for HDM injection. In the control group no significant changes in the LTC₄ concentrations were found over the 3-week time of the study. This pilot study showed that ketoconazole has anti-inflammatory properties and suggests that this drug may be effective in decreasing the skin concentrations of LTC₄ in atopic dogs. Résumé— Les effets de kétoconazole sur les résultats des tests intradermiques et la concentration en leucotriène C4 (LTC₄) dans la peau de chiens atopiques ont étéévalués dans une étude en double aveugle. Douze chiens atopiques sans dermite à Malassezia ont été selectionnés. Tous les chiens ont des tests intradermiques positifs aux acariens de la poussière de maison à 25 PNU mL^-1à 20 minutes. Six chiens ont reçu des comprimés de sucre, six autres ont reçu du kétoconazole à 5 mg kg^-1, 2 fois par jour par voie orale pendant 3 semaines. Aux jours 0 et 21, des injections intradermiques de liposaccharides (LPS), d'eau salée et d'aeariens de la poussière de maison sont réalisées et des biopsies des points d'injections sont effectuées 90 minutes après l'injection afin de mesurer la concentration en LTC₄ dans la peau. Les résultats des tests intradermiques ne sont pas modifies par la thérapeutique au kétoconazole. Par contre, le kétoconazole diminue significativement la concentration en LTC₄ induit par les injections intradermiques de LPS et d'eau salee. Le kétoconazole diminue également le LTC₄ induit par les acariens de la poussière de maison, mais il n'existe pas de différence significative entre les valeurs avant et après. La diminution moyenne de LTC₄ dans le groupe traité au kétoconazole est de 37% pour l'injection intradermique d'eau salée, de 42% pour l'injection intradermique de LPS et de 26% pour l'injection intradermique d'acariens de la poussière de maison. Dans le groupe de contrôle, aucune différence significative dans les concentrations en LTC₄ n'est observée durant les 3 semaines de l'étude. Cette étude démontre que le kétoconazole a des propriétés antiinflammatoires et suggèrent qu'il peut être efficace dans la diminution des concentrations en LTC₄ chez les chiens atopiques. [Marsella R, Kunkle, GA, Vaughn, DM, Macdonald, J. Double-blind pilot study in the effects of kétoconazole on intradermal skin test and leukotriene C₄ concentration in the skin of atopic dogs. (Etude en double aveugle sur les effets du kétoconazole sur les tests intradermiques et la concentration en leucotriène C₄ dans la peau de chiens atopiques.) Veterinary Dermatology 1997; 8 : 3–10.] Resumen Se evaluaron en un estudio piloto los efectos del ketoconazol sobre los resultados del test intradérmico cutáneo y sobre las concentraciones de leucotrieno C₄ (LTC₄) en la piel de perros atópicos. Se seleccionaron doce perros atópicos sin Dermatitis por Malassezia detectable. Todos los perros mostraban respuesta positiva inmedíata a la inyección del ácaro del polvo doméstico (HDM) a 25 PNU mL^-1. Seis perros recibieron una tableta control de azúcar y seis recibieron ketoconazol a dosis de 5 mg kg^-1 PO BID durante 3 semanas. En los días 0 y 21 se realizaron inyecciones intradérmicas de suero salino, lipopolisacárido (LPS) y ácaro del polvo doméstico, y se tomaron biopsias de las zonas inyectadas a los 90 minutos postinyección para cuantificar la concentración de LTC₄ en la piel. Los resultados de los tests intradérmicos no se afectaron por la terapia con ketoconazol. El ketoconazol disminuyó significativamente la concentración de LTC₄ que pudo haber provocado la inyección intradérmica de suero salino y LPS. El ketoconazol también disminuyó el LTC₄ inducido por el HDM pero las diferencias entre los valores previos y posteriores no fueron estadisticamente significativos. La disminución en la concentración medía de LTC₄ en el grupo de ketoconazol fue del 37% para la inyección de suero salino, 42% para la de LPS y 26% para la de HDM. En el grupo control no se encontraron alteraciones significativas en las concentraciones de LTC₄ durante las 3 semanas que duró el estudio. Este estudio piloto mostró que el ketoconazol posee propiedades antiinflamatorias y sugiere que este fármaco puede ser efectivo en disminuir las concentraciones cutáneas de LTC₄ en perros atópicos. [Marsella R, Kunkle, GA, Vaughn, DM, Macdonald, J. Double-blind pilot study in the effects of ketoconazole on intradermal skin test and leukotriene C₄ concentration in the skin of atopic dogs. (Estudio doble ciego sobre los efectos del ketoconazol en los tests intradérmicos cutaneos y en la concentración de leucotrieno C₄ en la piel de perros atópicos.) Veterinary Dermatology 1997; 8 : 3–10.] Zusammenfasung In einer Pilotstudie wurden die Wirkung von Ketoconazol auf Ergebnisse des intrakutanen Hauttests und der Leukotrien C₄ (LTC₄)-Konzentration in der Haut atopischer Hunde bewertet. Zwölf atopische Hunde ohne feststellbare Malassezia Dermatitis wurden ausgewählt. Alle Hunde hatten eine positive Sofortreaktion zu Hausstaubmilbenextrakt von 25 PNU mL^-1. Sechs Hunde erhielten eine Zuckertablette als Kontrolle und sechs Hunde erhielten Ketoconazol in einer Dosis von 5 mg kg^-1 oral zweimal täglich für drei Wochen. An den Tagen 0 und 21 wurden intrakutane Injektionen von physiologischer Kochsalzlösung, Lipopolysaccharid (LPS) und Hausstaubmilbenextrakt durchgeführt und 90 Minuten danach Biopsien an den Injektionsstellen entnommen, um die Konzentration von LTC₄ in der Haut zu messen. Ketoconazoltherapie hatte keinen Einfluss auf die Ergebnisse des Intrakutantests. Ketoconazol verminderte die LTC₄ Konzentration in den intrakutanen Injektionsstellen von physiologischer Kochsalzlösung und LPS. Die Konzentrationen des von Hausstaubmilbenextrakt-induzierten LTC₄ waren ebenfalls vermindert, die Unterschiede zwischen den Werten vor und nach der Injektion waren jedoch nicht statistisch significan. Die durchschnittliche Verminderung der LTC₄ Konzentration in der Ketoconazol-Gruppe betrug 37% für die Injektion mit Kochsalzlösung, 42% für die Injektion mit LPS und 26% für die Injektion mit Hausstaubmilbenextrakt. In der Kontrollgrupe wurden während der dreiwöchigen Studie keine signifikanten Veränderungen in der LTC₄ Konzentration festgestellt. Diese Pilotstudie zeigt die entzündungshemmende Wirkung von Ketoconazol und deutet darauf hin, daß dieses Medikament geeignet sein könnte, die Hautkonzentration von LTC₄ in atopischen Hunden zu verringern. [Marsella R, Kunkle, GA, Vaughn, DM, Macdonald, J. Double-blind pilot study in the effects of ketoconazole on intradermal skin test and leukotriene C₄ concentration in the skin of atopic dogs. (Doppelblind-Pilotstudie über die Wirkung von Ketoconazol auf den intrakutanen Hauttest und die Konzentration von Leukotrien C₄ in der Haut atopischer Hunde.) Veterinary Dermatology 1997; 8 : 3–10.]     

An investigation of the pathology of Mycoplasma mastitis in the cow

Summary

A field case of mastitis in cows, caused by Mycoplasma agalactiae var. bovis, formed the occasion to conduct an infection experiment. Five lactating heifers were infected in the udder at different times. The cows were slaughtered 2, 5, 7, 9 and 12 days p. i. and the pathological changes were studied. The investigation indicated that the pathological picture differed with time: in the acute stage, the inflammation was characterized by exudation of mostly eosinophils in the alveoli; later on, the mastitis was identified by an interstitial reaction with eosinophils and mononuclear cells, including plasma cells and lymphocytes; in the chronic stage, progressive fibroplasia around ductuli and alveoli, with hypertrophy of alveolar epithelium, was characteristic. The pathological findings are discussed.     

Comparison of three monoclonal and three polyclonal antibodies in the immunohistochemical diagnosis of canine autoimmune skin diseases

The aim of this study was to evaluate the utility of three monoclonal antibodies (mAbs), and two anticanine IgG and one anticanine IgM polyclonal antibodies (pAbs) for the immunohistochemical diagnosis of canine autoimmune skin diseases. Skin biopsies from 11 cases of pemphigus (7 foliaceus, 3 vulgaris and 1 erythematosus), 12 cases of discoid lupus erythematosus (DLE) and 12 cases of chronic hyperplasic dermatitis were used. The CA4E7 mAb (IgG1 + IgG2) showed similar sensitivity, but higher specificity and lower background than the two anti-IgG pAbs for the immunohistochemical diagnosis of pemphigus and DLE. The CA4F1 mAb (IgG2) and CA3H1 mAb (IgG2) showed moderate and low interepithelial reactivity, respectively, in autoimmune skin diseases, but strong staining of the cytoplasm of plasma cells of the inflammatory infiltrates. These results suggest that the CA4E7 mAb may be valuable in the immunohistochemical diagnosis of such disorders.     

犬瘟热病毒疫苗变异株的分离及其P1蛋白的表达、鉴定

从河北某宠物医院病犬的脾脏中分离到1株犬瘟热病毒,H基因测序结果表明,该分离株与疫苗株的同源性为99.5%,有3个氨基酸突变,但不影响其糖基化位点。将该株病毒P1基因克隆到pET32a(+)载体,转化大肠杆菌BL21(DE3)及Rosetta感受态细胞,在IPTG诱导下获得大小为50 ku,可溶性表达的P1重组蛋白。Western blotting、间接ELISA试验结果显示表达产物能被犬瘟热高免血清所识别,表明其具有良好的反应原性,有望用于犬瘟热的检测。     

A Serine12Stop mutation in PB1-F2 of the 2009 pandemic (H1N1) influenza A: a possible reason for its enhanced transmission and pathogenicity to humans

As the scientific community scrambles to define the ancestry and lineages of the eight segments of new pandemic H1N1 strain, we looked for unique genetic events in this virus''s genome to explain the newly found enhanced virulence and transmissibility among humans. Genome annotations of this virus identified a stop mutation replacing serine at codon 12 (S12Stop) of the PB1-F2 protein, a virulence factor in influenza A viruses. Here, we discuss the significance of this finding and how it may contribute to host specialization, explaining the virtual absence of the H1N1 influenza A virus strain in pig populations. This finding is expected to lead to a better understanding of the transmission and pathogenesis of the 2009 pandemic strain.     

Preliminary investigation into the prevalence of mucormycosis in the platypus (Ornithorhynchus anatinus) in three catchments in north-west Tasmania

Objective To investigate the distribution and prevalence of mucormycosis in platypus (Ornithorhynchus anatinus) from the Inglis, Emu and Black-Detention catchment areas in north-west Tasmania. Procedure A field study was performed and resulted in the examination of 44 wild platypuses; in addition, one dead platypus and two live platypuses were examined after they were independently submitted to a local veterinary clinic. Results No cases of mucormycosis were conclusively diagnosed. One platypus with signs consistent with those previously described in cases of mucormycosis was captured in the Emu River catchment. However, laboratory tests did not provide a definitive diagnosis for the lesion. Two platypuses from the Inglis catchment area had signs very similar to those previously described in cases of mucormycosis, but laboratory tests found Corynebacterium ulcerans to be the likely cause of the cutaneous ulcers on one of these platypuses and an unidentified fungal agent to be the cause of a cutaneous nodule in the other. Conclusions These findings do not prove that mucormycosis is absent from the populations studied. However, they may indicate that the prevalence of disease is low. The possibility that Mucor amphibiorum is present in a subclinical form in platypuses, or infecting another reservoir, is not excluded. The findings also suggest that caution should be exercised when diagnosing mucormycosis based on clinical findings alone and raise the possibility that some cases may have been incorrectly diagnosed.     

An investigation into the detection of the pulse in conscious and anaesthetized dogs

ObjectivesTo record the success rate of veterinary professionals and students at identifying the pulse in conscious and anaesthetized dogs. To explore the influence of clinical experience, pulse location, anaesthesia and likely confounding variables on the success of pulse palpation.Study designProspective, observational, randomized study.AnimalsA total of 54 client-owned dogs scheduled for general anaesthesia.MethodsFor each dog, three participants (senior anaesthetist, anaesthesia resident/nurse, veterinary student/animal care assistant) attempted pulse palpation at three locations (femoral, radial and dorsal pedal pulse) in conscious and anaesthetized dogs. The time to pulse palpation was measured with a stopwatch for each attempt and data were modelled using a multivariate Cox regression survival analysis (significance p < 0.05).ResultsThe overall success rate of pulse palpation was 77%, with a median time of 10.91 seconds (interquartile range 9.09 seconds). Success rate was lower in conscious dogs (67%) than in anaesthetized dogs (87%). There was a 77% lower likelihood of success at the radial than at the femoral pulse [hazard ratio (HR) 0.23, 95% confidence interval (CI) 0.38–0.69, p < 0.001]. Veterinary students/animal care assistants had a 71% lower likelihood of success than senior anaesthetists (HR 0.29, 95% CI 0.22–0.39, p < 0.001). Age, weight and American Society of Anesthesiologists physical status had no significant influence. Premedication/anaesthetic drugs, heart rate or mean arterial pressure had no significant influence on the time to pulse palpation in anaesthetized dogs. The median time to palpation was less than 10 seconds for all experience groups at the femoral location.ConclusionsPalpation of the femoral location had the greatest likelihood of success with the least amount of time. Monitoring the femoral pulse during induction of anaesthesia is suggested as a method for confirming spontaneous circulation. Pulse palpation improves with clinical experience.     

An investigation into the bacterial contamination of goniolenses in use in clinical practice

Objectives To report the incidence and evaluate the clinical significance of goniolens bacterial contamination in clinical use in dogs with three different usage protocols and one with an added cleaning protocol. Animals Studied and Methods Three groups of twenty dogs undergoing gonioscopy at a private practice in the UK had the goniolenses swabbed for bacteriology culture and identification prior to placement on the cornea. Three protocols of lens use, with 2 different types of goniolens, were studied. One protocol was then repeated with 21 dogs with a lens cleaning protocol prior to storage. Results Low levels of bacterial contamination were found in all 3 initial groups (10‐15%). No correlation was found between usage protocol used and rate of contamination and no correlation was found between length of storage between use and contamination. All bacteria cultured were considered naturally occurring commensals for the canine eye and environment. The group with a cleaning protocol had a 4.7% contamination rate. This was not statistically different from the non‐cleaning groups. Conclusions The rate of bacterial contamination of goniolenses in clinical practice is low and the bacterial contaminants consist of commensal bacteria, unlikely to be of detriment to the eye. Minimal contamination of the goniolenses was found and this did not appear to be of clinical significance. The introduction of a simple cleaning protocol did not produce a statistically significant reduction in bacterial contamination.