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Cytological Examination: Part II Clinical Pathology Microscopic Evaluation • Scan at 10x – Determine if stained adequately – Any localized or increased cellular areas – Scan for any large objects: parasites, foreign bodies, etc. – Look at what type of cells are found • Examine at 40x – Evaluate individual cells • Examine at 100x (oil immersion) – Cell morphology, nucleus, chromatin, cytoplasm Interpretation • Inflammatory vs. Non-inflammatory – Most important judgement – May not get a definitive diagnosis, but may be able to get a general idea/general process. – Cytology may be helpful on what’s the next diagnostic step to take • Culture • Biopsy • Rads • Serology Inflammatory cells found in Cytology • Neutrophils – May resemble same as blood neutrophils. – Be degenerative – Hypersegmented • Lymphocytes • Plasma cells – Active lymphocytes that have a very basophilic cytoplasm • Eosinophils – Mast cells- allergic inflammation • Macrophages – Large tissue monocytes. – Have abundant blue cytoplasm with vacoules that may contain phagocytized cells or debris – Oval to pleomorphic nucleus – Mesothelial cells • Cells that line the pleural, peritoneal,a nd visceral surfaces. • A type of macrophage Neutrophils in Tissue Lymphocytes and Plasma cells in tissue Eosinophils in tissue Mast cells in tissue Macrophages in Tissue Mesothelial Cells in Tissue Classifications of Inflammation • • • • Purulent Pyogranulomatous Granulomatous Eosinophilic • May also be classified as duration: – – – – Acute Subacute Chronic-active Chronic Purulent Inflammation (Abscess) • Most common type of inflammation • Usually caused by bacteria • Also called suppurative inflammation • Over 70% neutrophils with a few macrophages and lymphocytes Pyogranulomatous inflammation • Also referred to as chronic/active • Consists of macrophages and 50-75% neutrophils Granulomatous Inflammation • Greater than 50-70% of cells are mononuclear (monocytes, macrophage, giant cells). • Few neutrophils • Also called chronic inflammation. Eosinophilic Inflammation • Consists of greater than 10% eosinophils • Allergic related • May see a few mast cells, plasma cells and lymphocytes Selected Infectious agents of Cutaneous lesions • Bacterial agents – Tend to produce lesions characterized by >85% neutrophils, few macrophages, lymphocytes, and plasma cells. – Rods, cocci – Cytology is helpful in determining what kind of culture or stain is needed. • Fungal agents – Tend to have more macrophages than bacterial lesions, but may be mixed (pyogranulomatous). Low numbers of lymphocytes. • Sporothrix schenkii • Histoplasma capsulatum • Blastomyces dermatidis • Crytococcus neoformans • Coccidiodes immitis Sporiotrichosis: Sporothrix schenkii • Organisms are round to oval or cigar shaped • Stain pale to medium-blue cytoplasm with a slightly eccentric pink or purple nucleus. • Dimorphic fungus found in the environment worldwide • Inoculated into tissue via puncture wounds • Suppurative to pyogranulamatous • Skin lesions are characterized by multiple, non-painful, nonpruritic nodules that may ulcerate and drain purulent exudate. • Dissemination is rare Sporotricosis continued • Diagnose via cytology, biopsy, fungal culture • Easier to diagnose in cats, tend to have more organisms • Infected cats are highly contagious to humans • Treatment includes long term antifungals – Ketoconazole – Itraconazole • Prognosis is fair to good, but relapse is possible. Histoplasma Capuslatum • Round to oval- yeast-like • Dark blue/purple staining nucleus surrounded by a thin halo • Causes systemic disease • Cutanous lesions are rare, causes lungs or GI tract infections • Most common in termperate and subtropical areas. • Diagnosed through cytology, histopathology, fungal cultures, rads Blastomyces dermatidis: Blatomycosis • Caused by inhaling the conidia • Causes a disseminated infection – – – – Lymph nodes Skin Bones Other organs • Found in mostly acidic soils • Diagnosed by cytology, histopathology, serology and fungal cultures. • Most are single, blue, spherical and thick walled. • Pyogranulamatous Cyptococcus Neoformans: Cryptococcosis • Found worldwide • Organism is inhaled and establishes infections in the nasal cavity, sinuses, skin and other organs • Spherical, yeast-like organisms • Thick, clear mucoid capsule • May be budding or non-budding • Cats: URI signs, SQ swelling over bridge of nose, non-painful, may have CNS signs • Dogs: CNS signs and ophthalmic signs usually occur. Nodules on lips and nose. Coicidiodes Immitis: Coccidiomycosis • Dimorphic fungus and soil saprophyte endemic to desert areas • Organisms are inhaled and disseminate in body • Skin lesions are nodular, abscesses and draining tracts • Painful lameness Coccidiomycosis Continued • Spherical with thick deeply stained wall. • Diagnosis thought cytology, pyogranulamatous, histopathology, serology and fungal culture. • Treated by long term systemic antifungals (8-12 months) • Prognosis is unpredictable • Relapses are common • Fungal cultures are contagious • Infected animals are not considered contagious Leishmania donovani: Leishamaniasis • • • • • • • • • • • • Protozoa transmitted by blood-sucking sandflies Endemic to Central and South America Sporadic infections in the US A visceral and cutaneous disease that develops over months-years Lesions are dark and small to large and ulcerated. Diagnose by imprints, scraping and FNA Organism usually found in macrophages Small, round to oval Has a very light blue cytoplasm, an oval nucleus, and a small dark kinetoplast Usually numerous organisms found Not curable Contagious to other dogs through vector Non-inflammatory Lesions • Neoplastic – Epithelial – Mesenchymal (spindle cell) – Discrete Round cell tumor • Non-Neoplastic – – – – – – Cysts (sebaceous) Hyperplasia (prostatic hyperplasia) Dysplasia Hematomas Seromas Salivary Mucocele Epithelial Neoplasms • Tend to exfoliate cells in sheets or clumps • Cells tend to be large with moderate to abundant cytoplasm • Benign epithelial tumors – Papilloma – Epidermal inclusion cyst (epithelioma) – Perianal gland adenomas • Malignant epithelial tumors – Perianal gland adenocarcinoma – Squamous cell carcinoma Mesenchymal tumors: Spindle cell tumors • Tend to exfoliate individual cells instead of clusters • May be difficult to differentiate from normal granulation tissue (spindle cells are plump). • Difficult to differentiate from the different types of tumors on cytology. • Benign forms: – Fibromas – Lipomas – Hemangioma • Malignant forms: – Fibrosarcoma – Liposarcoma – Hemangiosarcoma Discrete Round Cell Tumors • Tend to exfoliate small to medium sized cells. • Also called cutaneous round cell tumors • Types: – Mast cell tumors – Cutaneous lymphosarcoma – Histioctyomas Transmissable venereal tumor Evaluation of Malignant Potential (Criteria of Malignancy) • • • • • • • • Variation of cell size Variation in nuclear size Multinucleated Increased nucleus: cytoplams ratio Mitotic figures Variation in nucleolar size/shape Coarse Chromatin pattern If more than 3 criteria are recognized in a high percentage of cells, this is strong evidenc for malignancy • If 1-3 criteria are present, may be either benign or malignant and should be sent to pathologist or biopsied. Submission of Cytologic Slides • Send 2-3 air-dried unfixed smears and 2-3 stained smears • Fluid samples should have smears prepared from them immediately • Also send EDTA and red top tubes filled with fluid • Mail in protective containers • Timely transportation service • Easy accesible and easy to collect cytology • Tranquilization/anesthesia seldome needed for sample collection • Quick-sample can be prepared, stained, and microscopically evaluated in minutes.