Vet Tech Clin Path Exam 1

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Clinical pathology

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The examination of diseased tissues, fluids or other materials from a living patient, using all of the techniques available including chemistry, hematology,enzymology, cytology, microbiology, parasitology, protozoology, immunology and histopathology.

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RVT's role in a veterinary laboratory can entail many things:

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Quality control of machinery

Installing software updates

Calibration of instruments

Inventory of testing supplies

Obtaining samples

Reading microscope and reporting results

Insuring timely and accurate results

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Reasons to perform clinical pathology assays?

Why run tests?

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Establish a diagnosis, rule out a disease, pre-anesthetic screening, monitoring effects of chronic medications on organ function, establishing a base-line prior to initiating medication,

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hematopoiesis

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The production of blood cells and platelets including erythropoiesis, leukopoiesis and thrombopoiesis.

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Erythropoiesis

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production of erythrocytes (red blood cells)

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Erythrocytes

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Red blood cells

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leukopoiesis

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Production of leukocytes (white blood cells)

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Leukocytes

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White blood cells s

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thrombopoiesis

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Production of thrombocytes (platelets)

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Thrombocytes

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Platelets

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Morphology

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The size, shape, and structure of an organism or one of its parts.

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Organelles

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A differentiated structure within a cell, such as mitochondrion, vacuole, or chloroplast, that perform at performs a specific function

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Anemia

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A pathological deficiency in the oxygen-carrying component of the blood, measured in unit volume concentrations of hemoglobin, red blood cell volume, or red blood cell number.

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Hemoglobin

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The iron-containing respiratory pigment in red blood cells of vertebrates, the functional unit of a red blood cell

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fibrinogen

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A protein in the plasma that is essential for the coagulation of blood and is converted to fibrin by the action of thrombin in the presence of ionized calcium

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Globulin

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A general term for proteins that are insoluble in water or highly concentrated salt solutions but soluble in moderately concentrated salt solutions.all plasma proteins except albumin and prealbumin are globulins.

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Reticulocyte

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An a nuclear, immature erythrocyte. Requires staining with a supravital stain to identify such as New Methylene Blue

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erythrocyte indices

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Calculated values that provide the average volume and hemoglobin concentrations of erythrocytes in a peripheral blood sample.

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Hypoxia

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Broad term meaning diminished availability of oxygen to the body tissues

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-Penia

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Suffix for decreased number of cells in the blood

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-Philip or -cytosis

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Suffix for increased numbers of cells in the blood

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Left shift

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Increased numbers of immature neutrophils (bands) in the blood

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Leukemia

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Neoplastic cells in the blood or bone marrow (not to be confused with feline viral leukemia)

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Leukemia response

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Marked leukocytosis (>50,000/microliter), usually the result of inflammatory disease, rather than leukemia

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anisocytosis

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Variation in RBC size

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polychromasia

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Variation in RBC color (staining) due to variations in hemoglobin concentrations.

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Polycythemia

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Increased number of circulating erythrocytes

Also from dehydration

Also known as 'hemoconcentration' when reading the PCV

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polycythemia

pathologic causes

Physiologic causes

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Pathologic causes: bone marrow disease, sever cardiac disease

Physiologic causes: stress

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Anemia

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Decreased oxygen-carrying capacity of blood

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Anemia can be due to:

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1. Decreased production of RBCs

2. Increased destruction or loss of RBCs

3. Decreased hemoglobin concentration within RBCs such as Iron deficiency anemia

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Anticoagulants:

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1. Ethylendiamine Tetracetic Acid

2. Heparin

3. Citrates and Oxalates

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Purple topped tube

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ethylendiamine Teracetic Acid (EDTA)

Preferred for hematologic studies because it does not alter cell morphology

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Green-topped tube

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Heparin

Used for tests requiring plasma samples (esp. Blood chemistry analysis)

Cannot be used for differential (WBCs don't stain)

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Blue or gray topped tube

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Citrates and Oxalates

Used for coagulation tests & transfusions

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Plasma

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The fluid filled portion of whole blood in which cells are suspended (anticoagulant was used)

90% water and 10% dissolved constituents- proteins, vitamins, carbohydrates, enzymes, lipids, salts, waste material, antibodies, etc.

Tubes yield ________ are EDTA, lithium heparin, etc.

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Serum

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This is plasma from which fibrinogen (a plasma protein) has been removed by the clotting process. Red-topped tube or serum separator tube is used

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Red-topped tube

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Contains serum (serum separator tube)

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What are some reasons to run a CBC?

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1. Diagnose a disease or pathologic process

2. Well animal screening/baseline

3. geriatric panel

4. Pre-anesthetic panel

5. Evaluate response to treatment of a chronic disease

6. excessive bruising or suspicion of a clotting disorder (low platelets)

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The Buffy coat

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The whitish/grey layer on top of RBCs after microhematocrit tube is spun down. Made of white blood cells and platelets.

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Erythrocyte morphology per species

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Canine- biconcave disc with central pallor

Feline- round with no central pallor

Avian and reptiles- nucleated RBCs

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Erythrocyte cell arrangement

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1. rouloux

2. Agglutination

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rouloux

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Grouping of RBC in stacks, like a stack of coins

Seen with increased fibrinogen or globulin concentrations

Seen in healthy horses and occasionally in normal cats

Can occur if slide is not made promptly or blood is refrigerated prior to slide preparation

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Agglutination

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Cells are clumped together

Can be seen in autoimmune disorders when antibiotics coat the cells and cause them to stick together

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anisocytosis

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variation in RBC size

(an- not, ISO- same, cyto- cell, oasis- condition of)

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macrocytes

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Usually young, polychromatophilic erythrocytes (reticulocytes).

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microcytes

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Small RBCs, possible Iron deficiency

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polychromasia

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Variable staining patterns, Basophilic, bluish RBCs when stained with Wright's type stain

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hypochromasia

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Decreased staining intensity due to low hemoglobin concentration.

This would be seen as a low MCHC. Common in Iron deficiency and usually have concurrent microcytosis (low MCV)

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Normochromic

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Normal staining intensity

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schistocytes

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RBC fragments, shearing of cell due to intravascular trauma (example is DIC, HSA)

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Crenation

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Normal artifact- shrinking of RBCs usually near the edge of the feathered edge. Can be due to improperly filled EDTA tube or slow drying of slide

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spherocytes

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Darkly staining red cells with reduced or no central pallor (detectable in dogs, not other species, because dogs normally have a pronounced central pallor)

Suggest immune mediated destruction of RBCs, hemolytic anemia (partial phagocytosis) or mismatched transfusion

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erythrocyte inclusion

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Are things that are inside or on the surface of the erythrocyte

Nucleated RBCs. Basophilic stippling

Howell-Jolly bodies. Heinz bodies.

reticulocytes

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Correct white blood cell count

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WBC count x100/ nRBCs +100

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Corrected reticulocyte count

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Reticulocyte count x (anemic animal PCV/normal PCV)

If greater than 1.5% is considered regenerative

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2 ways to classify anemia

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Bone marrow response

RBC size and hemoglobin concentration (RBC indices)

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Regenerative

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Nucleated RBCs, reticulocytes (greater than 1.5%) blood loss or blood cell destruction

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non-regenerative

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Zero or very few (<1%) reticulocytes

bone marrow dysfunction

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Normocytic

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Usually secondary to chronic diseasedd

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Macrocytic

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Regenerative (reticulocytosis)

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Microcytic

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Iron deficiency

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Hypochromic

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reticulocytosis

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Normochromic

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Most other types of anemia

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White blood cells are also called

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Leukocytosis

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Types of leukocytes:

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Myeloid line: neutrophils (granulocyte), monocytes, eosinophil (granulocyte), basophils (granulocyte)

Lymphoid line: lymphocytes

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4 different WBC abnormalities (all occur in neutrophils

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1. Nuclear hyposegmentation

2. nuclear hypersegmentation

3. Toxic change

4. Smudge cells

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Types of toxic change:

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cytoplasmic Basophilia

Dohle's bodies- ribosomes seen in toxic neutrophils

Vacuoles (foaminess)

intensely-stained primary granules (rare)

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The smallest blood "cell"

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platelets

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If platelet count is low it is called

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Thrombocytopenia

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If platelet count is high it is called

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thrombocytophilia or thrombocytosis

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Reasons to evaluate bone marrow

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1. Persistent, unexplained pancytopenia, neutropenia, neutropenia or thrombocytopenia

2. Cases of non-regenerative anemia

3. staging of neoplastic disorders (has cancer spread to bone marrow)

4.evaluation and diagnosis of leukemia

5.diagnosis of specific parasitic diseases.

6.immature or abnormal morphologic features of blood cells in systemic circulation on a blood film

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Sites used for bone marrow aspirates

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Head of humerus

Iliac crest

Femoral canal

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Bone marrow smears are classified as

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hypocellular (hypoplasia), acellular (aplasia), or hypercellular (hyperplasia)

Based on proportion of blood cells to fat

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Acute anemia:

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When >30% of marrow cells are blast cells.

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Chronic anemia:

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When <30% of marrow cells are blast cells

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myelofibrosis

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Most of the bone marrow is replaced by scar tissue and is incapable of producing blood cells

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Hemostasis

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The ability of the body's systems to maintain the integrity of the blood and the blood vessels. Very Simply stated, this is the ability of the blood to clot

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Preferred anticoagulant for coagulation test is?

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Sodium citrate (blue topped tube)

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Coagulation test

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Platelet count activated clotting time

Whole blood clotting time.

buccal mucosal bleeding time

Clot retraction time. Fibrinogen determination

one-stage prothrombin time test (OSPT)

Activated partial thromboplastin time (APPT)

PIVKA. D-dimer and fibrin degradation products sodium citrate

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Activated clotting time (ACT)

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Test requires a grey-topped Vacationer tube containing diatomaceous earth which activates the coagulation pathways.

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buccal Mucosa Bleeding Time (BMBT)

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Tests for abnormalities in platelet function

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one-stage prothrombin time (OSPT) test

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Prolonged time could be associated with severe Liver disease (liver produces clotting factors), DIC, hereditary or acquired deficiencies. Sensitive to vitamin K deficiency or antagonism (warfarin toxicity, rodenticide ingestion).

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Activated partial thromboplastin time (APTT)

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Tests intrinsic clotting mechanism, an automated assay. Also know as PT/PTT

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PIVKA

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An acronym for proteins induced by vitamin K absence. Vitamin K is required to activate factors II, VII, IX and X. this test is useful for differentiating between rodenticide toxicity from primary hemophilia when ACT is prolonged

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D-dimer and fibrin degradation products (FDPs)

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Useful in identification of DIC and provide diagnostic information in cases of liver failure, trauma and hemangiosarcoma.

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What is the most common inherited coagulation disorder of domestic animals is?

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Von Willebrand's disease, the result of decreased or deficient production of von Willebrand's factor

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The most common non-inherited coagulation disorder seen in small animal veterinary practice is?

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Thrombocytopenia

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DIC (disseminated intravascular coagulation)

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____ is associated with trauma, heat stroke, infectious disease and a variety of other serious illnesses

This disorder manifests as systemic hemorrhage or microvascular thrombosis

Poor to grave prognosis

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MCV

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mean corpuscular volume

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MCH

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Mean corpuscular hemoglobin

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MCHC

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Mean corpuscular hemoglobin concentration

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Most common problem encounter with a centrifuge?

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Improper balancing

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Purpose of refractometer?

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To indicate amount of solid in solution

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Spectrophotometry

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Measuring the amount of light transmitted or absorbed through a solution. It does not measure the refraction of light like a refractometer, but the amount of light that passes through the solution

measure the amount of light absorbed by the blood sample and give a numerical value. The value is then compared against the most common "norma" ranges for each blood constituent

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CBC machine measures actual cell counts for__________

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WBC, RBC, and platelets

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3 types of hematology analyzers we discussed

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Impedance, laser based, quantitative Buffy coat analysis

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3 layers of a spun down microhematocrit tube

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Packed RBCs, Buffy coat, plasma

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Pre- analytical errors

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Incorrectly entering species, age, breed

Sampling errors- lipemia (fat in serum), hemolysis (ruptured red blood cells in sample)

Labeling errors- incorrect patient or sample

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Analytical errors

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Improper maintained equipment random errors

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Post- analytical errors

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Clerical errors, transcription errors

Try not to lose outside lab results