Canine nodular dermatophytosis (kerion): 23 cases

Dermatophytosis is a common zoonotic disease, and one of its clinical presentations in the dog is nodular dermatophytosis (kerion). Because the infection is located within the dermis, routine diagnostic tests such as a Wood's lamp examination, microscopic examination of hair shafts for fungal elements and fungal culture can yield negative results. In such cases, histopathological examination with routine and special stains (periodic acid-Schiff, Gomori methenamine silver) is required to confirm the diagnosis. Nodular dermatophytosis in 23 dogs of different breed, age and sex with single or multiple nodules is described. Twelve dogs had a single nodule, and 11 dogs showed multiple lesions. Wood's lamp examination was negative in all cases. Microscopic examination of plucked hairs showed arthrospores in 8 of 23 cases. Skin scrapings in mineral oil looking for arthrospores and/or hyphae were positive in 12 cases. Impression smears of exudates were diagnostic in 21 of 23 cases (91%), showing arthrospores within fragments of hair shafts or free among neutrophils and macrophages (pyogranulomatous inflammation). Histopathology was performed in two cases. Fungal culture was positive for Microsporum canis in 16 dogs and for Microsporum gypseum in one dog. In six cases, the causative agent was not identified by fungal culture. All dogs were treated with systemic antifungal therapy and in eight cases with concurrent antibiotic therapy. Nodular dermatophytosis resolved in all dogs with the prescribed treatments within 4 to 8 weeks. Transmission to people or other pets in the home was not found.     

Genetic immunization with the immunodominant antigen P48 of Mycoplasma agalactiae stimulates a mixed adaptive immune response in BALBc mice

A DNA vaccine against contagious agalactia was developed for the first time, encoding the P48 of Mycoplasma agalactiae. Specific immune responses elicited in BALB/c mice were evaluated. Both total IgG and IgG1 were detected in mice vaccinated with pVAX1/P48. Proliferation of mononuclear cells of the spleen, levels of gamma interferon, interleukin-12, and interleukin-2 mRNAs were enhanced in immunized animals. Results indicate that pVAX1/P48 vaccination induced both T_h1 and T_h2 immune responses. Nucleic acid immunization could be a new strategy against M. agalactiae infections and may be potentially used to develop vaccines for other Mycoplasma diseases.     

Pancytopenia in a cat with visceral leishmaniasis

Abstract: A 4‐year‐old, domestic shorthair, female spayed cat was presented for decreased appetite and depression. Severe pancytopenia with erythrocyte autoagglutination was found. The cat was seronegative for feline immunodeficiency and leukemia viruses. Immune‐mediated hemolytic anemia was suspected but no response to treatment with a blood transfusion, enrofloxacin, and prednisone was observed. Blood and bone marrow smears obtained 11 days later contained Leishmania amastigotes in the cytoplasm of neutrophils and macrophages, respectively. Serologic and PCR testing of peripheral blood confirmed infection with Leishmania infantum. Despite treatment, the cat worsened clinically and was euthanized. At necropsy, visceral dissemination of the parasite was confirmed. The findings in this case indicate that visceral leishmaniasis should be considered as a differential diagnoses in cats with pancytopenia in areas endemic for Leishmania. In addition, amastigotes may be observed in peripheral blood neutrophils.     

Extra hepatic expression of the acute phase protein alpha 1-acid glycoprotein in normal bovine tissues

Combined quantitative (real-time) polymerase chain reaction and immunohistochemistry were used to evaluate the expression of the minor acute phase protein alpha 1-acid glycoprotein (AGP, orosomucoid) in bovine extrahepatic tissues. AGP was produced mainly in the salivary glands and spleen, whereas minor expression was detected in all other tissues sampled, including lung, lymph nodes, uterus, ovary, kidney and tongue. The findings were consistent with immunohistochemical results. In view of the immunomodulatory and direct antibacterial activity of AGP, its expression in the salivary glands may signal an involvement in the regulation of the local immunity, even in non-pathological conditions.     

Evaluation of static telepathology in veterinary diagnostic cytology

BACKGROUND: Telepathology is the process of diagnostic pathology performed on digital images viewed on a display screen rather than by conventional light microscopy with glass slides. At present, electronic mail (e-mail) attachment is the most common communication medium for telepathology, because it is relatively simple and incurs minimal cost. OBJECTIVE: Our objective was to test whether telediagnosis through Internet e-mail could be applied to veterinary diagnostic cytology. METHODS: Using 20 cytology cases, on which a consensus diagnosis was reached by 3 experienced pathologists, a total of 130 images were digitized and sent as e-mail attachments from the Unit of Anatomic Pathology of the Faculty of Veterinary Medicine of Naples to the Unit of Pathology of the Faculty of Veterinary Medicine of Messina, Italy. The images were reviewed at the Unit of Pathology of Messina, where the consulting pathologist formulated his diagnoses by visualizing the images on the monitor of a computer. The telediagnoses were compared with the consensus diagnoses and with the consulting pathologist's interpretation using conventional light microscopy at a later date. RESULTS: The median time to capture images was approximately 30 minutes for each case. The median time to make a telecytodiagnosis was a few seconds for each case. Overall, there was good agreement (85%) between the consensus diagnosis and the consultant's telediagnosis. In 100% of the cases there was agreement between the consulting pathologist's telediagnosis and conventional glass slide diagnosis. CONCLUSION: Telepathology by e-mail provides acceptable efficacy and a faster turnaround time than post and can be applied to veterinary diagnostic cytology.     

Flow cytometric patterns in blood from dogs with non-neoplastic and neoplastic hematologic diseases using double labeling for CD18 and CD45

BACKGROUND: In dogs, flow cytometry is used in the phenotyping of immunologic cells and in the diagnosis of hemic neoplasia. However, the paucity of specific antibodies for myeloid cells and B lymphocytes and of labeled antibodies for multicolor techniques limits the ability to detect all leukocyte subpopulations. This is especially true for neoplastic and precursor cells. CD18 and CD45 are expressed on all leukocytes and are involved in cell activation, and together could be useful in helping determine cell lineage. OBJECTIVES: The purpose of this study was to double label canine blood for CD18 and CD45 and to use the differential expression of antigens to identify leukocyte populations in dogs with non-neoplastic and neoplastic hematologic diseases. METHODS: A template was developed using blood samples from 10 clinically healthy dogs and a back-gating technique. Differential leukocyte counts obtained with the template were compared with those obtained by manual and automated methods on blood samples from 17 additional healthy dogs. Blood samples obtained from 9 dogs with non-neoplastic (reactive) hematologic diseases and 27 dogs with hemic neoplasia were double stained for CD18 and CD45 using mouse anticanine CD18 monoclonal antibody (mAb) plus phycoerythrin-conjugated rat anticanine CD45 mAb and fluorescein isothiocyanate-conjugated rabbit antimouse IgG. Hemic neoplasms were diagnosed by cell morphology, and immunophenotypic and cytochemical markers. RESULTS: With the double label, neutrophils, eosinophils, monocytes, and T- and B-lymphocytes were identified. In reactive disorders, a population of activated neutrophils with high CD45 and CD18 expression was detected. In hemic neoplasia, cell lineage was easily determined, even in acute leukemia. CONCLUSIONS: Double labeling for CD18/CD45 may be useful as a screening method to evaluate hematologic diseases and help determine cell lineage, and to aid in the selection of a panel of antibodies that would be useful for further analysis.     

Localisation of iron in wheat grain using high resolution secondary ion mass spectrometry

Insufficient iron (Fe) is one of the most prevalent micronutrient deficiencies in humans, with billions of people affected. Cereal grains are an important source of Fe for humans but the bioavailability of Fe in cereals is generally low. Information regarding the cellular and sub-cellular localisation of Fe in wheat grain will aid optimising nutrient delivery for human health. In this study high resolution secondary ion mass spectrometry (NanoSIMS) was used to map the distribution of Fe in the aleurone layer and in the endosperm of immature wheat grain. Iron was shown to be localised strongly in the phytin globoids in the aleurone cells and to a lesser extent in the cytoplasm around the starch granules in the endosperm.     

Trafficking and deposition of prolamins in wheat

The processing properties of wheat flour are largely determined by the structures and interactions of the grain storage protein, the so called “gluten proteins”. The synthesis, folding and deposition of the gluten proteins take place within the endomembrane system of the plant cell. Several studies have suggested that two trafficking pathways occur in wheat, with the proteins either being transported via the Golgi apparatus into the vacuole or assembling within the ER to form protein bodies that are subsequently internalized into vacuoles by a process analogous to autophagy. The precise routes of trafficking and deposition of individual gluten proteins remain unclear and so are the molecular and cellular mechanisms regulating their sorting in the two pathways.     

Seasonal stability in parasite assemblages of the Brazilian flathead, Percophis brasiliensis (Perciformes: Percophidae): predictable tools for stock identification

A comparison of the composition and structure of parasite communities of the Brazilian flathead, Percophis brasiliensis Quoy et Gaimard (Perciformes: Percophidae) among seasons during one year was carried out in the Argentine Sea. A total of 195 fish specimens were examined and 25 parasite species were found. Parasite communities in seasonal samples showed a high degree of homogeneity in taxonomic composition and infection levels. Similarity analysis showed that the seasonal stability within and between samples was constant in both the composition and community structure throughout the year. Parasites can, therefore, be considered predictable markers for fish stock identification, independently of the season of capture, at least on an annual scale.     

Antimycotic effectiveness against dermatophytes: comparison of two in vitro tests

Seven antimycotic drugs (econazole, enilconazole, fluconazole, griseofulvin, itraconazole, ketoconazole, and miconazole) were tested against 36 dermatophyte strains (19 M. canis, 7 T. mentagrophytes, 5 M. gypseum, 2 M. cookei, 1 T. rubrum, 1 T. ajelloi, and 1 T. terrestre) isolated from animals, humans, and the environment. Two in vitro methods were compared: a micro-dilution test based on the CLSI M38-A method, and a disk-diffusion test. Fluconazole was not effective in vitro against the dermatophytes. Econazole and enilconazole were the most effective. Thirteen strains were griseofulvin-resistant. The correlation between the two methods was statistically significant for enilconazole, griseofulvin, itraconazole, and miconazole.