Pulmonary adiaspiromycosis in the Eurasian beaver (Castor fiber) inhabiting Poland.

Adiaspiromycosis is a rare fungal infection caused by saprophytic fungi Emmonsia spp. (type Ascomycota) occurring especially in small free-living mammals. The aim of this study was to evaluate the occurrence of histopathological lesions asscociated with adiaspiromycosis in the Eurasian beaver inhabiting Poland. In order to evaluate the presence of natural adiaspiromycosis we systematically investigated beaver populations from north-eastern Poland for adiaspores in the lungs. This study reveals for the first time the presence of pulmonary adiaspiromycosis of Eurasian beaver in Poland. As far as we know, there is no published data regarding pulmonary adiaspiromycosis in human patients in Poland.

Immunosuppression in Dogs During Mammary Cancer Development.

Cancer immunosuppression that facilitates tumor progression and metastasis evolves by development of an immunosuppressive network. The aim of this study was to assess this network in dogs with benign or malignant tumors with or without confirmed metastasis. The authors showed that the number of various T cell subpopulations was constant during tumor development; however the number of regulatory T cells (Tregs) was significantly higher in tumor-bearing dogs than in healthy individuals. The number of myeloid-derived suppressor cells (MDSCs) and their p-STAT3 expression (which is a negative regulator of hematopoiesis and regulates VEGF expression) were higher in cancer patients than in control dogs, however their number increased significantly in late-stage cancer patients. Canine mammary carcinomas with confirmed metastases to either lymph nodes or internal organs had greater MDSCs and Treg infiltration than benign mammary tumors or malignant mammary tumors for which metastases had not been detected. Similarly, expression of p-STAT3 and VEGF-C was the highest in tumors with confirmed metastases. This research shows changes occurring in the blood (n = 30 patients) and tumor tissue of patients (n = 100) during canine mammary tumor development. The findings should be considered preliminary because of the small number of samples. Nonetheless, the findings suggest that a high level of Tregs and MDSCs as well as high expression of p-STAT3 and VEGF-C may significantly contribute to mammary tumor progression and metastasis in dogs.

Multidrug therapy of Mycobacterium avium subsp. avium infection in experimentally inoculated budgerigars (Melopsittacus undulatus).

The aim of the study was to determine whether the four-month experimental therapy of mycobacteriosis in budgerigars may cause a complete recovery. A group of nine budgerigars was infected with a Mycobacterium avium subsp. avium isolate with proven pathogenicity for budgerigars. Five weeks post-inoculation, multidrug therapy was started. Another group comprising six birds received the same treatment but no infection, and the third group also comprising six birds was kept without infection or treatment as a control. The adopted antibiotic regimen included clarithromycin 61 mg/kg b.w., moxifloxacin 25 mg/kg b.w. and ethambutol 60 mg/kg b.w. administered by crop gavage every 12 h for 18 weeks. Despite a significant improvement in the condition of the infected, treated birds, the four-month therapy was not sufficient for the complete recovery of all.

Histological Evaluation of Selected Organs of the Eurasian Beavers (Castor fiber) Inhabiting Poland.

There is a general scarcity of data on the histological structure of major organs in the Eurasian beaver (Castor fiber). This study presents the histological characteristics of beaver organs such as the liver, spleen, cardiac muscle, lungs and kidneys. Tissue samples were collected from 21 beavers and analysed. Selected samples of tail tissue were additionally examined. Tissue samples were placed in neutral buffered formalin and embedded in paraffin. 4-µm-thick sections were stained with haematoxylin and eosin, and other staining techniques were also used. Scant amounts of inter-lobular connective tissue were found in the liver. Ion or copper deposition was not observed, but scattered cytoplasmic glycogen deposits were present in hepatocytes. Our results suggest that beavers have defensive rather than storage spleens. Interestingly, the presence of melanin in splenic red pulp was noted. The histological structure of the examined organs closely resembled that of other rodent species. According to our knowledge, this is the first study evaluating the histological structure of beaver organs. Nevertheless, precise characterization of the evaluated organs requires further work with the involvement of accurate and reliable techniques, such as molecular biology or electron microscopy methods.

Hematological parameters in relation to age, sex and biochemical values for mute swans (Cygnus olor).

The knowledge of the correct morphological and biochemical parameters in mute swans is an important indicator of their health status, body condition, adaptation to habitat and useful diagnostic tools in veterinary practice and ecological research. The aim of the study was to obtain hematological parameters in relation to age, sex and serum biochemistry values in wild-living mute swans. We found the significant differences in the erythrocyte count, hematocrit, hemoglobin concentration and erythrocyte sedimentation rate in relation to age of mute swans. There were no differences in hematological values between males and females. The leukogram and H/L ratio did not vary by age and sex in swans. Among of biochemical parameters the slightly increased AST, ALP, CK, K, urea, decreased CHOL and TG values were recorded. As far as we know, this is the first study in which the morphometric parameters of blood cells in mute swans were presented. We found extremely low concentration of lead in blood (at subthreshold level). No blood parasites were found in blood smears. The analysis of body mass and biometric parameters revealed a significant differences dependent on age and sex. No differences in the scaled mass index were found. Our results represent a normal hematologic and blood chemistry values and age-sex related changes, as reference values for the mute swan.

Cutaneous paraneoplastic syndrome in a cat with thymoma.

Feline cutaneous paraneoplastic syndrome is a rare disorder associated mainly with pancreatic carcinoma and thymoma. In this report the authors describe the case of a 12-year-old cat with paraneoplastic exfoliative dermatitis associated with thymoma. Lateral radiographic examination of the chest showed a small subtle soft tissue density in the ventral part of the first and second intercostal space, which together with skin changes suggested thymoma. Because of pain associated with the skin condition, costs of treatment and the risk associated with surgical treatment, the owner chose euthanasia of the cat. Post-mortem examination revealed a tumour which was diagnosed as thymoma by histopathological examination.

High agreement of routine cytopathology and immunocytochemistry in canine lymphomas.

The aim of this study was to compare the concordance of immunophenotype established with routine cytopathology (slides stained with Giemsa solution) and immunocytochemistry according to CD3 and CD79 alpha immunoreactivity. The study was performed on cytological samples of 70 canine lymphomas subtyped on the basis of the updated Kiel classification system. Additionally, cytologic samples were examined immunocytochemically for the CD3 and CD79 alpha antigens presence and thus immunophenotype of neoplastic growth was confirmed. The cytopathological and immunocytochemical diagnoses were then compared; in order to measure the concordance between immunocytochemistry (IC) result and Giemsa stain result of the same sample the Cohen's kappa coefficient was calculated. On the basis of the results of immunocytochemistry of 70 cases of canine lymphoma examined 42 were recognized as B cell lymphoma and 28 as T cell lymphoma. Full accordance between the results of routine cytopathology and IC was obtained in 63 out of 70 examined dogs (90% of cases). It can be concluded that cytopathological examination of Giemsa stained smears is helpful in determining the lymphoma cells immunophenotype. Additionally, it seems that combination of routine cytopathology and immunocytochemistry in cases of canine lymphomas allows to obtain the precise diagnosis in 90% of cases, and allows to receive most important information that is necessary for planning of appropriate therapy and to determine prognosis. Finally, this routine procedure allowed to eliminate the need of collection of tissue samples during surgery or core-biopsy and thus time, cost and patient discomfort related to more complex and invasive medical procedures can be easily reduced.

Derivation of feline vaccine-associated fibrosarcoma cell line and its growth on chick embryo chorioallantoic membrane - a new in vivo model for veterinary oncological studies.

Feline vaccine associated fibrosarcomas are the second most common skin tumor in cats. Methods of treatment are: surgery, chemotherapy and radiotherapy. Nevertheless, the usage of cytostatics in feline vaccine associated sarcoma therapy is limited due to their adverse side effects, high toxicity and low biodistribution after i.v. injection. Therefore, much research on new therapeutic drugs is being conducted. In human medicine, the chick embryo chorioallantoic membrane (CAM) model is used as a cheap and easy to perform assay to assess new drug effectiveness in cancer treatment. Various human cell lines have different tumors growth on CAM. In veterinary medicine such model has not been described yet. In the present article derivation of feline vaccine associated fibrosarcoma cell line and its growth on CAM is described. The cell line and the tumor grown were confirmed by histopathological and immunohistochemical examination. As far as we believe, this is the first attempt to create such model, which may be used for further in vivo studies in veterinary oncology.

Cytopathological diagnosis of visceral histiocytic sarcoma in five dogs.

Histiocytic sarcoma is a malignant neoplastic proliferation of atypical histiocytes with tendency to spread, characterized by fast progression to disseminated form--disseminated histiocytic sarcoma. Cytopathology is a low, invasive, cheap, and quick method of diagnosis commonly used in veterinary oncology. The aim of the presented study was description of cases of visceral histiocytic sarcomas in dogs diagnosed by cytopathology and immunocytochemistry. The study was conducted on 5 dogs which were brought to the veterinary clinic because of unspecific clinical signs and tumoral masses recognized in the thoracic or abdominal cavity. Samples of cells were collected during ultrasonography-assisted fine-needle aspiration biopsy (FNAB), smears were stained with Giemsa method and immunocytochemistry (CD3, CD79alpha, cytokeratin, vimentin, desmin) was also performed in all patients. Four of five dogs were Bernese mountain dogs, nonspecific clinical signs of systemic disease were present in all cases. Visceral mass or masses were detected by ultrasonography or radiography. Final diagnosis of histiocytic sarcoma was obtained on the basis of routine cytopathological examination and confirmed by immunocytochemistry. On the basis the results obtained it can be stated that in cases of typical clinical and cytopathologic pictures, examination of cellular samples collected during ultrasonography-assisted fine-needle biopsy supported by some immunocytopathological characteristics seems to be sufficient method of diagnosis of histiocytic sarcoma in dogs. Visceral histiocytic sarcoma should be included into differential diagnosis in every Bernese mountain dog with nonspecific clinical signs, ambiguous results of hematologic examination and when tumoral mass or masses within a body cavity were detected in imaging techniques.

Histopathology in diagnosis of broiler chicken and layer diseases--review of cases 1999-2010.

The aim of this study was to estimate the prevalence of histopathological lesions in the different organs in relation to the commercial-type and the age of birds (i.e. broiler chickens and layers). During the period 1999-2010 a total of 189 cases was submitted to the Division of Animal Pathomorphology, Department of Pathology and Veterinary Diagnostics at WULS. Most cases were found in broiler chickens (66.7%). The majority of the histopathological lesions were detected in the liver and lymphoid organs. In of 29% cases of hepatic injury pathognomonic lesions associated with inclusion body hepatitis (IBH) were found. The mean age of birds was 23 days. Among IBH cases proventriculitis (58%) was more often found than gizzard lesions (25.8%). Interestingly, we noted some intranuclear inclusions in the epithelial cells within the proventriculus. A low percentage of histopathological evidence of infectious bursal disease (IBD) was reported in chickens. The gastrointestinal tract was the second most frequent predilection site for histopathological lesions. Histopathological findings within the heart and lungs were less common and were more often seen in the upper respiratory tract. Cases of infectious laryngotracheitis (ILT) were registered in broiler chickens (3.2%, mean age 37 days) and in layers (4.8%; mean age 196 days). Lesions associated with Marek's disease, avian leukosis and fowl pox were recognized only in layers, respectively in 3.2% (mean age 176 days), 1.6% (mean age 205 days) and 1.1% (mean age 196 days) of all cases. Avian encephalomyelitis (AE) was noted only in 0.5% of all cases.

Relationship between receptors for insulin-like growth factor- I, steroid hormones and apoptosis-associated proteins in canine mammary tumors.

In the veterinary literature there are few data concerning the expression of insulin-like growth factor type I (IGF-IR) in the canine mammary gland tumors. The aim of the present study was the evaluation of IGF-IR expression and its correlation to the expression of estrogen receptor alpha (ERalpha) and progesterone receptor (PR), proteins: Bcl-2, Bax, p53 in canine mammary gland tumors, and also a correlation with other features: bitch's age, tumor diameter, histologic type of tumor, degree of histologic malignancy, proliferate activity. The study was done on 112 epithelial neoplasms: 21 (19%) were adenoma, 38 (34%) complex carcinoma (adenocarcinoma), 47 (42%) simple carcinoma (adenocarcinoma) and 6 (5%) solid carcinoma. Histochemistry and immunohistochemistry methods were employed. It was shown that more common and/or higher IGF-IR expression in cells of canine mammary gland tumors was related to the histologic type of cancer of worse prognostic (solid and simple carcinoma), high histologic degree of malignancy (III degrees) but the statistical analysis did not reveal any significant differences. We observed the high degree of IGF-IR expression in tumors which displayed the high ERalpha and PR expression. These results suggest the involvement of IGF-IR in the development of hormonosensitive canine mammary tumors. Additionally, the significant positive correlation between expression of IGF-IR and p53, Bax was found. Our study provides some evidence that interactions exist between the IGF-IR and these apoptosis-associated proteins may contribute to the development and progression of canine mammary gland tumors. These results require further investigations.

Density of tumor-associated macrophages (TAMs) and expression of their growth factor receptor MCSF-R and CD14 in canine mammary adenocarcinomas of various grade of malignancy and metastasis.

Several years ago, the presence of macrophages in the tumor microenvironment was thought to be an inflammatory response to kill the cancer cells. Now, this is clear that the inflammatory cells that exit blood vessels and migrate to the tumor tissue play an important role in cancer progression. Various cells present in the tumor microenvironment enhance cancer growth and invasiveness by secretion of tumor-enhancing products. That is why tumors should not be treated as only aggregates of cancer cells but as separate structures. Macrophages form a major component of the inflammatory infiltration in tumors, where they are termed tumor-associated macrophages (TAMs). To the best of our knowledge, up-to-date there were no studies on tumor associated macrophages and the role of the tumor microenvironment in tumor invasion/metastasis in dogs. This is the first study performed to asses if the number of TAMs and expression of MCSF-R (macrophages colony stimulating factor receptor) and CD14 (LPS co-receptor) are associated with the grade of tumor malignancy and its ability to metastasize. We have performed immunohistochemical analysis of 50 canine mammary adenocarcinomas of various grade of malignancy (1st, 2nd, 3rd) and tumors that gave local or distant metastases. The results indicate that in dogs, similarly to humans and mice, the number of tumor associated macrophages is related to the cancer ability to metastasize. Our results also indicate that the expression of MCSF-R and, what is particularly new finding, CD14 is associated with tumor malignancy and its ability to metastasize. Hence, these molecules play a role in tumor progression, metastasis and microenvironment interactions. These results show that in dogs we should treat the tumor as a whole organ rather than just try to eliminate the cancer cells.

Gene expression pattern in canine mammary osteosarcoma.

Canine mammary sarcomas are usually very aggressive and easily metastasize. Unfortunately the biology of this type of tumor is not well known because they are a very rare type of tumors. The aim of this study was to find differences in gene expression patterns in canine mammary osteosarcomas (malignant) versus osteomas (benign) using DNA microarrays. Our microarray experiment showed that 11 genes were up-regulated in osteosarcoma in comparison to osteoma whereas 36 genes were down-regulated. Among the up-regulated genes were: PDK1, EXT1, and EIF4H which are involved in AKT/PI3K and GLI/Hedgehog pathways. These genes play an important role in cell biology (cancer cell proliferation) and may be essential in osteosarcoma formation and development. Analyzing the down-regulated genes, the most interesting seemed to be HSPB8 and SEPP1. HSPB8 is a small heat shock protein that plays an important role in cell cycle regulation, apoptosis, and breast carcinogenesis. Also SEPP1 may play a role in carcinogenesis, as its down-regulation may induce oxidative stress possibly resulting in carcinogenesis. The preliminary results of the present study indicate that the up-regulation of three genes EXT1, EIF4H, and PDK1 may play an essential role in osteosarcoma formation, development and proliferation. In our opinion the cross-talk between GLI/Hedgehog and PI3K/AKT pathways may be a key factor to increase tumor proliferation and malignancy.