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Hematology Test #2
Terms in this set (222)
Most primitive form of a cell line; can renew themselves, differentiate and mature
Bone Marrow Pool
Cells go through maturation process in the bone marrow and are stored until needed
Leukocytes 2 categories
Granulocytes and Agranulocytes
Neutrophil, Eosinophil and Basophil
Monocytes and Lymphocytes
Production of granulocytes
Granulocytes seen in circulation represent approx. ___ of the body's granulocytes (___ are in the bone marrow)
40% and 60%
How long do granulocytes stay in circulation?
Where are granulocytes eliminated?
In the liver, respiratory tract and GI tract
How long do granulocytes stay in the tissue?
What is the lifespan of a granulocytes?
Large, round irregular cell; High N:C ratio; 2-3 nucleoli seen; Smooth basophilic cytoplasm
Type II Myeloblast
Large # of azurophilic primary granules ; granules made of lysosomes (digest proteins), enzymes, antibacterial agents; Nucleus occupies 1/2 of cell
- Neutrophilic: Oval, round nucleus, light blue cytoplasm, faint pink granules
- Eosinophilic: More basophilic cytoplasm, distinct large orange granules
- Basophilic: Smaller, dark blue, few, large black granules
Nucleus becomes kidney shaped; Chromatin more condensed/clumped; Cytoplasm background less blue; Granules are neutrophilic, eosinophilic, basophilic
Nucleus becomes thinner, horseshoe shaped; has parallel sides; twist to conform to cytoplasm spaces
Horseshoe shaped nucleus, rounded ends or S shaped; Indentation greater than 1/3 width of nucleus is segmented neutrophil even if only 2 lobes; Left shift occurs if # of band neutrophils are high; Toxicity common in left shifts; Seen in inflammatory conditions
Mature Granulocytes Neutrophil Canine
Cytoplasm pale pink or light blue; Dense nuclear chromatin; Small, poorly stained, pink granules
Mature Granulocytes Neutrophil Feline
Cytoplasm pale pink or light blue; Dense nuclear chromatin; Often lacks visible granules
Mature Granulocytes Neutrophil Bovine
Pink-tinted cytoplasm due to pink granules
Mature Granulocytes Neutrophil Equine
Slightly pink cytoplasm; Often lacks visible granules
Mature Granulocytes Neutrophil
Final stage - polymorphonuclear cell; Nuclear membrane not smooth, irregular widths; segments into 2 to 5 lobes always connected by nuclear filaments; Cells with greater than 5 lobes are considered hypersegmented
Lifespan of a Neutrophil
Function of a Neutrophil
Phagocytize and digest fungi and bacteria
Absolute increase in neutrophils; due to extreme exercise, stress, chronic infection and steroid therapy
Absolute decrease in neutrophils; due to drugs, shock, decreased production in bone marrow, hemolysis, acute infection, congenital defect
Mature Granulocytes Eosinophil
Nuclei of eosinophils usually have 2 lobes; granules vary in size, shape and number with species
Mature Granulocytes Eosinophil Feline
Light pink, rod-shaped granules are numerous (may cover nucleus); Small amount of faint bluish cytoplasm
Mature Granulocytes Eosinophil Canine
Orange-red, round granules vary in size and number; May occasionally have vacuoles
Lifespan of an Eosinophil
Function of Eosinophils
Regulation of allergic and acute inflammatory responses; May induce tissue damage, some phagocytic ability
Decrease in eosinophils; may be normal; due to emotional stress
Increase in eosinophils; chronic disease; histamine factors from mast cells; allergic reactions; parasitism; exercise; may occur in canine estrus
Basophilic cytoplasm and granules; rare, may be seen with eosinophilia
Mature Basophil Feline
Pale pink granules with a few dark purple granules
Mature Basophil Canine
Granules vary in #, size and intensity; may cover nucleus (appear as a vacuolated monocyte); has grey-blue cytoplasm
Lifespan of a Basophil
Function of Basophils
Exhibit chemotaxis and limited phagocytic ability; granules contain histamine (role in allergies) and heparin (inhibit coagulation -- important in inflammation); migrate towards foreign proteins
Increase in basophils; associated with increased eosinophils; chronic myelogenous leukemia, polycythemia
Too few to tell
What does a Monocytic Series consist of?
Monoblasts, promonocytes and monocytes
Resemble myeloblasts except nucleus is round to convoluted
Similar to myelocytes and metamyelocytes
Large cells; Variably shaped nuclei - kidney shaped, convoluted, band, or doughnut; chromatin more diffuse than neutrophil; Cytoplasm is typically blue-gray staining, abundant and vacuolated
Monocytes develop into ____________ when they leave the blood and enter the tissues
Macrophages are found where?
The spleen, bone marrow, liver, stomach, small intestine and at sites of inflammation
Function of monocytes/macrophages
Defense against intracellular parasites; Necrotaxis and necrophagocytosis; Present processed antigen to T lymphocytes; Release inflammatory mediators
Remove old and damaged RBCs
What do T lymphocytes do?
Help initiate the immune response
Increase in monocytes; need to clean blood or tissue of debris; sub-acute and chronic inflammation caused by bacteria, fungi or protozoa; severe crushing trauma; hemorrhage into tissues or body cavities; neoplasia; stress or corticosteroid therapy
not clinically significant
Where are lymphocytes produced?
Migrate to the lymphatic system which includes the lymph nodes, thymus and spleen; Varying sizes; Circulating lymphocytes decrease with age
Low # of lymphoblasts and prolymphocytes seen; Difficult to differentiate from rubriblasts and prorubricytes; Nucleus almost fills the cell; Categorized into small, medium or large, depending on amount of cytoplasm; some may be reactive; usually present and very diagnostic of lymphomas
Small # circulate in blood; Mostly reside in lymphoid organs; Nuclei are round and can be slightly indented; Nucleus eccentric, chromatin coarse and clumped; Cytoplasm light blue; Atypical lymphocytes contain granules; have chromocenters
Produce antibodies, develop immunity to infections; Initiate an immune response; Active in tissue rejection
Decrease in lymphocytes; Corticosteroid or stress induced; Loss of lymph; Drugs, ionizing radiation or neoplasia; viral disease; severe bacterial infection
Increase in lymphocytes; Physiologocal or epinephrine induced; Chronic disease; Leukemia/lymphosarcoma; Hypoadrenocorticism; Excited or frightened cats; recovery from viral or bacterial infection
Increased WBC count due to increased muscle activity with corresponding increase HR and RR; Could be seen in stressful situations, exercise, pregnancy; lactation and post-prandially
Reflects systemic stress and participation of white cells in fighting underlying causes
Where can changes be seen with WBCs
Nucleus, cytoplasm, granules or all three
Abnormalities of Neutrophils
Nucler Hypersegmentation, Nuclear Hyposegmentation, Pelger-Huet Anomaly
Neutrophils with 5 or greater lobes
Absolute increase in hypersegmented neutrophils
Causes of Nuclear Hypersegmentation
Old cells, Chronic inflammation or infection, Anemia, Steroid use
- Left Shift; may also see toxic cytoplasm, donut shaped nuclei and giant sized neutrophils
- Cells vary from bands to metamyelocytes and myelocyes
Increased # of immature neutrophils in the blood
Cause of Nuclear Hyposegmentation
Seen in inflammatory conditions
Congenital defect causing hyposegmentation of granulocytes and monocytes; Cells are mature; Nucleus has condensed chromatin can take on band or metamyelocyte nuclear shape
Barr Bodies (sex chromatin "drumsticks")
What % of neutrophils in healthy female animals will have Barr bodies?
Mimics PH anomaly but fewer hyposegmented cells seen; Response to severe infection, sometimes reaction to certain drugs
What is Toxic Neutrophils associated with?
Conditions such as inflammation, infection and drug toxicity
Why may toxic neutrophils occur?
Due to accelerated production of cells in response to disease
Toxic Neutrophil changes include what?
Dohle Bodies, Cytoplasmic Basophilia, Vacuolization, Pyknosis, Karyorrhexis, Karyolysis and Toxic Granules
Coarse, irregular gray to blue cytoplasmic inclusions (RNA) that can be seen single or in pairs; Common in felines as a toxic change
Where are Dohle Bodies found?
Infections, poisoning, burns and following chemotherapy
Blue cytoplasm usually seen with vacuoles; Slightly more severe sign of toxicity
Can be recorded as mild to severe; can be seen in lymphocytes as well; gives cytoplasm a frothy appearance; Artifact if prolonged exposure to EDTA
Condensed or shrunken nucleus; Very dense chromatin; May fragment (karyorrhexis)
Represents cell death; Nucleus becomes swollen and nuclear membrane degrades
Less common; Cytoplasmic granules become very prominent
Broken WBCs; Lacy/net-like cells; May have crisscross basket weave pattern; Can also show nuclear remnant lacking intact cytoplasm
What is Basket/Smudge Cell fragility due to?
- Excess anticoagulant
- Rough handling
- Extended period of time from collection to processing
WBC Infectious Inclusions
Bacterial, fungal, parasitic and viral
Sometimes contaminant; Clinically significant if neutrophils contain phagocytized bacteria
Cytoplasmic inclusion of lymphocytes, monocytes, neutrophils and platelets; Appears as small basophilic clusters in the cytoplasm
How is Ehrlichia spp transmitted?
How is Ehrlichia spp best diagnosed?
Buffy coat smears as it is usually present in small #
Clinical signs of Ehrlichia spp?
Pyrexia, inappetence, lethargy
Gram negative bacteria transmitted through ingestion of infected snails and other water insects
Causes of Neorickettsia risticii
Potomac Fever in Horses
Who can get infected by Neorickettsia risticii?
Dogs and Cats
What does Neorickettsia risticii infect?
Granulocytes, monocytes and macrophages
Similar appearance to Ehrlichia canis
How is Anaplasma phagocytophilia transmitted?
Who does Anaplasma phagocytophilia affect?
Equine neutrophils and eosinophils
Anorexia, lethargy, difficult ambulation
Dimorphic fungus found in soil containing bird and bat feces
Histoplasmic capsulatum: Cytoplasmic inclusions
Neutrophils, eosinophils, monocytes and macrophages
Histoplasmic capsulatum infection is caused by?
Aerosol contamination of the respiratory tract, sometimes GI contamination
Who does Histoplasmic capsulatum affect?
People, dogs, cats, sheep, pigs and horses
Signs of Histoplasmic capsulatum
Hepatosplenomegaly, pulmonary disease, lymphadenomegaly, anorexia, depression, conjunctivitis, optic neuritis, unresponsive pyrexia
Hepatozoon canis transmitted by?
Through eating ticks, not from being bitten by one
Cytoplasmic inclusions in neutrophils and monocytes; Appear as large oblong structures
Signs of Hepatozoon canis
Anorexia, pyrexia, pale mm, lethargy, diarrhea, lameness, purulent ocular discharge
Canine Distemper (CDV)
Contagious, incurable, often fatal, multi-systemic viral disease that affects the respiratory, GI, and central nervous systems
Canine Distemper commonly transmitted by?
Through bodily secretions that become airborne viral particles that the dog breathes in
Signs of Canine Distemper
Pyrexia, discharge from eyes and nose, depression, anorexia, diarrhea, pneumonia, and vomiting
What does Canine Distemper develop?
Large aggregates of viral particles seen in RBCs and WBCs; Transient
Appear as unstained rods within cytoplasm
How is Mycobacterium spp confirmed?
Who does Mycobacterium spp affect?
Cattle, pigs, sheep, deer, poultry, horses, birds, rats, cats, and goats
Leishmania infantum transmitted by?
What does Leishmania infantum cause?
Severe systemic disease
Signs of Leishmania infantum
Cutaneous lesions, muscle wasting, lymphadenopathy, ocular lesions, epistaxis, anemia, renal failure, diarrhea, lameness, normal or decreased body temperature
Who does Leishmania infantum affect?
Humans, dogs and rodents
They are engulfed by macrophages in which they transform and divide into amastigotes
Cytauxzoon felis signs
Depression, weakness, dyspnea, dehydration, pyrexia and icterus
What can you see with Cytauxzoon felis?
Splenomegaly, hepatomegaly, enlarged lymph nodes, renal and pulmonary edema
Forms within RBCs, lymphocytes and macrophages
Vector of Cytauxzoon felis
Rare inherited disease; abnormally stored substances in cell; blue to magenta granulation in cytoplasm
What is lysosome storage due to?
Birman Cat Granulation
Fine eosinophilia to magenta granules in neutrophils; inherited in Birman cats; differentiate with toxic granulation and lysosomes disorders
Neutrophils show lysosomes in cytoplasm & leg plump granules pink to purple in colour; tendency to bleed; abnormal platelet function
What is the sequence that platelets are produced in?
Megakaryoblast, promegakaryocyte, megakaryocyte and thrombocyte
What happens during development of Megakaryocytopoiesis?
Nucleus duplicates by mitosis
How does development of megakaryocytosis end?
With large polylobulated nucleated cell
Which is the largest cell in bone marrow?
Cytoplasm colour of cell of megakaryocytopoiesis
Intensely basophilic to granular light blue
What regulates the development of megakaryocytes?
Most immature form in the sequence; 1, 2 or 4 round nuclei; fine chromatin in nucleus; one or more nucleoli; basophilic and non-granular cytoplasm; High N:C ratio
Usually 2-4 nuclei seen; nuclear chromatin becomes more condensed as cell mature; moderately basophilic cytoplasm; can see azurophilic granules in some areas of the cytoplasm; N:C ratio is lower
Nuclei are joined in a lobulated mass; cytoplasm less basophilic and has numerous, small, azurophilic granules; huge cell
What magnification can you evaluate a megakaryocyte?
If megakaryocyte is dwarf in size with single or less lobulated nucleus, what could that indicate?
How many platelets can one megakaryocyte produce?
Thrombocytes; important part of hemostasis; provide initial coagulation factor; plug small ruptures in blood vessels; round to oval, anucleate structures
How are platelets formed?
fragmentation of megakaryocyte cytoplasm
What species is clumping of platelets common?
Average size of platelets?
⅓ to ½ the size of an RBC for a given species
aka Shift platelets; same size or larger than an RBC; early release from the bone marrow
Where are macroplatelets seen?
Myeloproliferative disorders and in certain breeds with hereditary platelet defects
Very small (less than ⅓ the size of a RBC)
Where are microplatelets seen?
Elongated cigar-shaped platelets suggest hemorrhage
Where can anisocytosis be seen?
Early bone marrow release
Have cytoplasmic projections; may form small clumps; partially activated platelets
Degranulation occurs from activation; appear light blue in projections; very few to no granules
Where is hypogranular seen?
Ehrilchia platys morulae; appear as large granules but are inclusions; tightly packed basophilic granules in cytoplasm
Prepare blood smear, perform estimated platelet count; can also perform estimated count with WBC differential
Formula for estimated count with WBC differential
Platelets/100WBC x total WBC count/uL = platelets/uL
Platelets/uL normal for dogs
Platelets/uL normal for cats
What do you document with a platelet evaluation?
Clumping, estimate # of platelets/oif, morphology (present of macro or microplatelets, aniocytosis, activation, presence of hypogranular platelets, parasites)
What are oher platelet evaluations
Automated analyzers, reticulated platelet count
What could thrombocytopenia be due to?
Collection circumstances, increased destruction, decreased production, increased consumption, loss due to hemorrhage,
Thrombocytopenia: Decreased platelet count
Abnormal platelet production usually accompanied by anemia and neutropenia, Accelerated platelet removal immune or non-immune, Abnormal distribution of platelets, combination of all.
Causes of abnormal platelet production
Drugs, infection, viral, bone marrow production
Causes of accelerated platelet removal immune or non-immune
Autoimmune, infection or bleeding disorder, neoplasia
Causes of abnormal distribution of platelets
enlarged spleen and toxicities
Thrombocytosis is due to
Life stage, Myeloproliferative disorder, Reactive thrombocytosis
Reactive Thrombocytosis is due to
Increased thrombopoietin stimulated by the release of inflammatory cytokines
Where is reactive thrombocytosis seen?
Cancer and inflammatory conditions
Thrombocytosis: Increased Platelet Count
Essential - primary bone marrow disorder, secondary to disease, physiological
Thrombocytopathia: Platelet function disorders
Acquired, abnormal production, dysfunction of normal platelets, congenital, deficiency of plasma factors, membrane and granular
Membrane and granular - abnormalities
Canine thrombasthenic thrombopathia, Basset hound thrombopathia, spitz thrombopathia, cocker spaniel bleeding disorders
BMBT, ACT, PT, PTT
Buccal Mucosal Bleeding Time (BMBT)
Tests platelet function, fast efficient test
When would you perform a BMBT test?
When animal is showing signs of a bleeding disorder such as: Petechia, Epistaxis, Hematuria
Prolonged BMBT due to:
Platelets defects, increased capillary fragility, Von Willebrand's Disease
Activated Clotting Time (ACT)
Tests intrinsic clotting mechanism, grey top tube
Prothrombin Time (PT)
Tests for extrinsic clotting mechanism, usually sent away to lab or done in-house special machine, test is sensitive to Vitamin K deficiency or antagonists such as Warfarin
What is the normal range for Prothrombin time test?
7 to 10 seconds
Prolonged results of Prothrombin time associated with
Severe liver disease, DIC, acquired or hereditary disorders
Partial Thromboplastin Time (PTT)
Tests intrinsic clotting mechanism, usually sent away to lab or done in-house with special machine
What is the normal range for partial thromboplastin time?
What is the abnormal range for partial thromboplastin time due to?
Acquired or hereditary disorders or heparin treatments
Bone Marrow Evaluation
Neutropenia, unexplained thrombocytopenia, non-regenerative anemia, neoplastic conditions, proliferative abnormalities, abnormal immunoproteins
Aspirate less accurate than core, administration of pain meds, pre-procedure, induction agent & local anesthetic needed, complications rare but consist of hemorrhage & infection, prepare site with surgical scrub
must have a removable stylet; 16-18 gauge needle between 1-1.5 in
Common collection sites in dogs and cats
Proximal humerus (greater tubercle), femur (trochanteric fossa) and ilium
Common collection sites in large animals
sternum, ilium & ribs
Are aspirate preps on euthanized animals poor quality?
When is a bone marrow core biopsy used?
When getting dry aspiration as result of technique or when marrow is packed with cells or in myelofibrosis or in very young animals even if highly cellular
Evaluation of smear in a healthy animal
rubricytes and metarubricytes usually comprise of 80-90% of the erythroid cells; metamyelocytes, bands and segmented myeloid cells comprise 80-90% of the myeloid cells
Cellularity of Sample: If more than 75% cells
Cellularity of Sample: If more than 75% fat
Hypercellular is response to what conditions?
Anemia, inflammation, neoplasia
Evaluation of RBCs in Bone marrow smears
Erythroids are smaller, more spherical & have condensed nuclei; basophilic cytoplasm becomes more polychromatophilic and them pink as mature
Evaluation of Granulocytes in Bone marrow smears
Larger, paler cells are usually granulocyte precursor; nucleus elongates as matures
Lymphocytes in bone marrow
small lymphocytes may be present
_______________ and ______________ are rare but when seen are in low numbers and hard to differentiate with RBCs
What indicates neoplastic leukemias and lymphomas or immune mediated responses called lymphoproliferative disorders?
High numbers of precursors
transformed lymphocytes; appear with antigenic stimulation
What do plasma cells have?
Abundant basophilic cytoplasm, round eccentric nucleus, clumped looking chromatin; more abundant cytoplasm than rubricytes; perinuclear clear zone
Monocytes and macrophages
Very small portion; difficult to differentiate with granulocytic cells
Monocytes - inflammation
Monoblasts - myeloproliferative disorder
Macrophages - when phagocytosis of cells is prominent, inflammatory and neoplastic and infectious
Naphthol AS-D acetate preparation; both monocytes and granulocytes contain esterase
Monocytes vs granulocytes
cells with heavy, blue granules are monocytes and cells with only a few blue granules are granulocytes
Osteoblast and osteoclast
giant cells that form and phagocytize bone; consistently found in young animals; rarely in adults except in bone sarcomas
giant, multinucleated, clearly separated rounf to oval, abundant blue cytoplasm, variably sized magenta granular material
What are osteoclast responsible for?
Lrgr than plasma cells, abundant vacuolated, basophilic cytoplasm, eccentric nucleus
What are osteoblast responsible for?
Iron stores in macrophages; appears gray to black; readily visualized with Romanowsky stains; easily found; increases with age
What species are hemosiderin absent in?
Bone marrow derived but rarely seen; lrg cell, round nuclei and lrg # of deep purple granules in cytoplasm
When do mast cells occur?
In varying amounts in immune response, inflammation, neoplastic conditions & anemia due to bone marrow problems & allergies
From vascular, connective and endothelial tissue as well as reticular matter; usually ruptured in smear preparation
Where do free nuclei come from?
Stromal and other bone marrow cells
Lymphoid precursors are called?
lacelike chromatin called basket cells
Nuclei in cells seen in reproduction are called?
What can bone marrow help with?
Find a diagnosis when combined with patient history, clinical findings, CBC and other diagnostic tests
Provide info about the pathogenesis of abnormalities found in blood
Neoplastic change in cells within bone marrow
Neoplastic changes are classified as:
Myeloproliferative disorders - increase in blast cells in blood and/or bone marrow for RBC, WBC, monocytes and platelets
Lymphoproliferative disorders - neoplastic proliferation of lymphocytes and plasma cells and are more common
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