Flow cytometric immunophenotyping of bone marrow in cats diagnosed with hematopoietic disorders

Cytological examination of bone marrow in cats, due to the large number of cells and various growth phases is somewhat complicated. The use of flow cytometric techniques and monoclonal antibodies are appropriate methods in the diagnosis of hematopoietic malignancies. The purpose of the present study is to determine cell-surface antigens for various developmental stages of feline bone marrow cells in hematopoietic disorders using flow cytometric. In this study, bone marrow cells from 4 cats with hematopoietic disorders and 2 clinically healthy cats, were labeled with 5 types of anti-feline MAbs included: CD21-like [Cr-Br], T lymphocyte subpopulation, CD-172a, Granulocyte, Pan-Leukocyte [CD45-like] and then analyzed using flow cytometric. The results revealed changes in immunophenotyping and light scatter properties compared with normal cases. The percentage of CD45, Granulocyte and CD172a markers in the bone marrow of a cat with erythroleukemia were lower compared with normal bone marrow. In a cat with myelodysplastic syndrome, scatter plot indicated an increase in the immature myeloid cells and a decrease in mature myeloid cells. It was concluded that cytological examination of bone marrow with studying dispersion studies on cells using flow cytometric and usage of a panel of antibodies such as CD21-like[Cr-Br], T lymphocyte subpopulation, CD-172a, Granulocyte, Pan-Leukocyte [CD45-like] could support the diagnosis of feline hematopoietic abnormalities

[ effects of age, sex and slaughtering on cellular distribution of bronchoalveolar lavage fluids in calves and cattle]

Based on our knowledge, there have been no studies about the effect of age, sex, lobe and slaughtering stress on cellular distribution of bronchoalveolar lavage fluids in calves and cattle in Iran. The main purpose of this study was to compare the cell distribution pattern of bronchial alveolar lavage fluids in calves and cattle in terms of age, sex, type of lung lobes and the stress of slaughtering. Bronchoalveolar lavage was performed in twelve calves at three different ages' groups [less than 2 months, 2-4 months and 4-6 months]. 250 milliliters sterile normal saline was infused through the tracheal tube and lavage was performed using syringe pressure. Post-mortem BAL was performed on twelve isolated lungs by infusing 150 milliliter normal saline. The lavage fluid was collected in sterile plastic tubes. The slide smears was prepared by pellet which were air-dried and stained with a Giemsa stain. Slides were evaluated cytologically. The data was analyzed by T-test, One Way ANOVA and post hoc Tukey test .Also, the Spearman correlation coefficient was used to assess the age and cell. Significant negative correlations were observed between cells such as epithelial cells, macrophages and mast cells, with a correlation coefficient of -0.430, - 0.059 and- 0.267- and age although the differences were not significant [p<0.01]. Statistically significant difference was observed in percentages of mast cells in males [1.48 +/- 1.25%] and females [zero percent] [p=0.04]. The differences were significant [p=0.019] between percentage of epithelial cells in the lungs of slaughtered cattle and lavaged lung of calves. The percentages of macrophages in slaughtered animals was significantly decreased [p=0.019] compared with live calves. Significant differences were not observed in cell density in different ages and sexes. But in the cell density in live animals [583 cells per microliter] was significantly higher than the lungs of slaughtered [237 cells per microliter] [p=0.03]. It seems that age and lobe do not affect pattern of lung immune cells. Sex and slaughtering stress, however may cause changes in immune cell type and density and lead to develop more respiratory disease

[Comparison of diagnostic accuracy of ultrasonography and radiology in radial critical-sized defects healing process in rabbit]

Healing of Critical-Sized Defects [CSDs] is one of the major challenges facing orthopedic surgeons. To assess the bone healing process usually plain radiography is used. Serial radiography results in certain side effects and recent findings are indicating the early detection of bone healing via ultrasonography. The purpose of current study is to compare the diagnostic accuracy of radiography and ultrasonography in healing process of radial CSDs in rabbit. Sixteen New Zealand White Rabbits were used in this study. The radial CSDs of 15 mm size were created in a routine surgical procedure described previously and the two diagnostic tools were compared 12 weeks post-surgery. In case of obtaining different results from radiology and ultrasonography, to determine which diagnostic imaging method is of more accuracy, the histopathologic results of samples from the same day were used. In 15 cases of 16, ultrasonography findings were confirmed by radiography and only in one case they were in contradiction, in which radiographs showed an Atrophic Nonunion while Ultrasound detected an outstanding filled defect with a reverberation artifact underneath. In histopathology, the defect was filled with an osteoid matrix of high cellularity and calcification was obvious in some regions, confirming the ultrasound results. This study indicates that in diagnostic imaging of CSDs, the ultrasonography and radiography are usually consonant and even ultrasound is more accurate than radiology. In addition, in case of detection of a reverberation artifact and lack of any healing-related finding in radiography, this artifact may be an indication of osteoid matrix formation

[Molecular study on Ehrlichia canis in thrombocytopenic dogs]

Infection with Ehrlichia canis, a gram negative obligatory intracellular bacterium, causes canine monocytic ehrlichiosis which is the worldwide disease in dogs. The objective of this study was to investigate the prevalence of E. canis in thrombocytopenic dogs using nested PCR and diagnostic role of thrombocytopenia in the infection. Blood samples collected from 40 dogs attended in Teaching small animal hospital of Tehran University were classified as group A [platelet counts below 101.000/microL, thrombocytopenic, n=11], B [101.000-200.000/microL, thrombocytopenic, n=15] and C [platelet counts more than 201.000/microL, non-thrombocytopenic, n=14] according to their platelet counts. 16S rRNA was analyzed by nested PCR using specific primers. 16S rRNA gene fragment of E. canis were detected in five samples of group A [45.5%], three samples of group B [20%], and one sample of group C [7.1%]. Prevalence rate of infection was statistically higher in group A than the other groups [p=0.02]. In total, approximately one third of thrombocytopenic dogs had demonstrable E. canis infection [30.7%]. While thrombocytopenia cannot be considered as specific marker for detection of E. canis infection, it can be used as a surveillance test prior to other diagnostic methods

Desmin expression by a gastrointestinal stromal tumor in a dog

This report describes the histological and immunohistochemical features of a small intestinal tumor that resembled a human gastrointestinal stromal tumor [GISTs] in a seven-year-old male cross-breed dog. This was unique because of the expression of desmin by the tumor. Grossly, the white-gray tumor measured 8.0x4.0x6.5 cm and was 760 gram in weight. It was cystic and enveloped the jejunum. Histopathologically, long spindle-shaped cells were arranged densely in the interwoven pattern. The tumor cells had a low rate of mitosis, were pleomorphic, and were positive for vimentin, alpha-smooth muscle actin, desmin, S100 and C-KIT and negative for CD34 on immunohistochemistry. The expression of desmin in the cytoplasm of this tumor cells is a rare event in these types of tumors

Immune-mediated hemolytic anemia in cats referring to Veterinary Teaching Hospital of Tehran [2006-2007]

Immune-mediated hemolytic anemia [IMHA] is characterized by the destruction of erythrocytes or sometimes bone marrow erythroid precursors mediated by immunoglobulins [IgG, IgM], with or without complement [C[3]]. The main objectives of this study were to assess the laboratory test results of IMHA and to investigate its possible underlying causes in cats referring to the Veterinary Teaching Hospital of Tehran. The Coombs' test [CT] was performed in 74 cats with PCV below 0.35. The test was positive in 26 cats. These positive CT cats were categorized into four groups based on the PCV ranges and type of anemia including: Group A: 9 of 26 cats had nonregenerative anemia [PCV median, 0.22] [1 feline leukemia virus positive which had erythroleukemia, 1 feline infectious peritonitis positive, 1 with Hemoplasma spp., 3 with renal failure, 2 with inflammatory disease and 1 with no diagnosis]. Group B: 4 cats [PCV median, 0.31] had a regenerative anemia with severe dehydration. Group C: the other 9 cats with a normal range of PCV [median, 0.34] involved with various conditions [vaccination, parturition, acetaminophen poisoning, osteoporosis, and renal failure]. Group D: the remaining 4 cats with a marginal range of PCV [median, 0.30] had a history of inflammatory disease and drug therapy. The results of this study indicated that infectious diseases and drug therapy were the main factors associated with positive Coombs' test results