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What are granulocytes?
Named by staining of nucleus...neutrophils, eosinophils, or basophils
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What are mononuclear WBC's?
Lymphocytes and Monocytes
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List three general methods of obtaining a leukocyte count from most accurate to least.
- Manual from Unopette
- Automated (from particle counter, expanded buffy coat, laser flow cytometry)
- Estimation techniques
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What are the functions of neutrophils?
- First line of defense in tissue
- Phagocytosis
- Anti-microbicidal action (kills bugs)
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How many nuclear segments does a normal segmented neutrophil contain?
3 - 5 divisions
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Describe the chromatin pattern of the segmented neutrophil
clumpy
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What is a band?
Immature neutrophil - not yet segmented
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Describe what a band looks like.
- Sides are parallel
- U or S shaped
- Smooth nuclear membrane
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Is there such a thing as a band eosinophil
no
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Define left shift.
- Neutrophils released from bone marrow early, in response to inflammation
- Once bone marrow gears up, it can release enough segs but early in the process when there aren't enough segs, the bands are sent out.
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Define regenerative left shift.
- There are more segs than bands
- Leukocytosis
- We know the bone marrow is working because there are more segs
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Define degenerative left shift.
- Bone marrow is not able to keep up with the demand
- Bands are = or > segs
- Leukopenia because the body just doesn't have any more
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What is degenerate neutrophil and what causes it?
- "Toxic changes" in Neutrophils
- Reaction of segs to inflammation, infection, toxemia
- Happens within the cytoplasm
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What is another name for toxic seg?
Degenerative neutrophil
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Describe what changes occur as toxic changes occur in neutrophils.
- Basophilic cytoplasm
- Cytoplasmic vacuoles/foaminess
- Dark blue granules: Dohle bodies
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What is hypersegmentation and what is it due to?
- More than 5 lobes
- Causes: an old seg, artifact from not making blood smear within 30 minutes
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Where are neutrophils produced?
bone marrow
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What is Pelgar-Huet anomaly?
All mature neutrophils look like bands
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List three abnormal blue blobs found within neutrophils
- Viral Inclusion Bodies
- Ehrlichia Canis (intracellular parasite)
- Histoplasm (fungal) Organisms
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Where are eosinophils produced?
bone marrow
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How long do eosinophils circulate?
30 minutes
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What is eosinophilia?
Increased number of eosinophils in blood
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What does the presence of eosinophilia indicate?
- Parasites (internal and external)
- Allergic reactions (hypersensitivities)
- Eosinophlic leukemia, some tumors (MCT - mast cell tumor)
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What is eosinopenia?
Decreased number of eosinophils in the blood
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What causes eosinopenia?
Glucocorticoids (cortisols) - Hyperadrenocorticism (Cushings), Iatrogenic, Stress (increase in natural cortisol)
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Are basophils commonly found on blood smears?
no
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When they are found, they often accompany what other WBC?
eosinophils
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What do mast cells look like? Where are they normally found?
- Many dark purple granules, may obstruct the view of the nucleus
- Found in tissue
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List the cells of granulopoeisis (in order of production)
- Myeloblast
- Promyelocyte
- Myelocyte
- Metamyelocyte
- Band
- Mature granulocyte
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List the cells of monocytopoeisis (in order of production).
- Monoblast
- Promonocyte
- Monocyte
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List the cells of lymphopoeisis (in order of production)
- Lymphoblast
- Prolymphocyte
- Lymphocyte
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How long does it take to go from myeloblast to segmented granulocyte?
10 - 11 days
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Name six tissues that contain a lot of lymphoid tissue.
- Lymph nodes
- Spleen
- Tonsils
- Thymus
- Mucosal surfaces
- Bone Marrow
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What is a plasma cell?
Activated B-lymphocytes (make lots of antibodies)
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Where do we find plasma cells?
- Lymph nodes
- Bone marrow
- Spleen
- Occasionally seen in blood
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What does a plasma cell look like?
Eccentric nucleus with perinuclear clear zone
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Lise five causes of lymphocytosis.
- Post-vaccination
- Chronic infections
- During infection recovery
- Lymphocytic leukemia
- Part of physiologic leukocytosis in cats (ventipuncture, restraint) - when they get excited or stressed their lymphocytes will go up
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List five causes of Lymphopenia
- Many viral diseases (example - parvo)
- Stress
- Glucocorticoids
- Immunosuppressive drugs (chemotherapy)
- Ionizing radiation
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Where are monocytes produced?
Bone marrow
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Where do macrophages come from?
Monocytes in blood transforms to macrophages in tissues
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What are the appropriate terms for increases and decreases in the different WBC cell types?
- Leukocytosis/Leukopenia
- Neutrophilia/Neutropenia
- Eosinophilia/Eosinopenia
- Lymphocytosis/Lymphopenia
- Monocytosis/Monocytopenia
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What is the primary site of lymphocyte production?
In bone marrow
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What causes stress leukogram?
Due to endogenous (came from the body) or exogenous (given to the body)
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Describe how excitement can affect the leukogram.
It flushes the marginated neutrophils from the capillary walls and will show an increase in segs
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How does excitement produce neutrophilia?
Excitement increases segs in circulating pool by "flushing" them from the walls of capillaries into the circulating blood
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List the causes for monocytosis.
- Chronic infections
- Fungal infections
- Monocytic leukemia
- Situations with an increase in cortisol (stress, hyperadrenocorticism)
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Cell-mediated immunity is performed by _____ and humoral immunity is performed by _____.
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How do you calculate absolute counts from relative WBC percentages?
% from each type of cell X WBC count = absolute count (cells/ul)
The % is the relative count
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What is the function of segs?
First line of defense in tissue
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What is the function of bands?
Help with inflammation
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What is the function of lymphocytes?
- T-Lymphocytes: Destruction of foreign invaders (viruses, bacteria, etc)
- B-Lymphocytes: Produces antibodies (helps flag invaders for the rest of the immune system to destroy)
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What is the function of monocytes?
Eats old RBC
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What is the function of macrophages?
Phagocytes in tissue
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What is the function of eosinophils?
- Attaches to and kills worms
- Attached by chemical mediators released by mast cells udring allergic reactions
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What is the function of basophils?
Mediator of inflammation
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Which WBC is the largest?
Monocytes
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What typically causes leukocytosis?
Neutrophilia
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What are the three different types of neutrophilia?
- Physiologic neutrophilia
- Inflammatory leukogram
- Stress leukogram
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What are the causes of neutrophilia?
- Excitement
- Inflammation
- Stress
- Neoplasia
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Describe what a monocyte looks like.
- Vesicular (lacey) chromatin
- Cytoplasm gray-blue
- Vacuoles in cytoplasm
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Where, besides the blood, do monocytes live?
- In tissue as macrophages
- In reticuloendothelial system as macrophages (Kupffer cells)
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In cats, what other cell is commonly seen in the peripheral blood as part of physiologic leukocytosis? And why?
lymphocytosis due to epinephrine release during a stressful situation (ex/ventipuncture)
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Is a stress leukogram caused by epinephrine release due to a difficult venipuncture?
yes
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What are the differences between an inflammatory leukocytosis and a physicologic leukocytosis?
Inflammatory Leukocytosis: Due to neutrophilia, neutrophilia with or without a left shift
Physicologic Leukocytosis: Due to epinephrine release which causes tachycardia which makes the blood pump faster and clushes the neutrophils from the capillary walls
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What are causes of leukopenia?
- Viral infection
- Overwhelming bacterial infection
- Gram negative endotoxemia
- Poor nutritional state (starvation)
- Physical agents (radiation)
- Chemical agents (chemotherapy)
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What are the causes of neutropenia?
- Overwhelming inflammation
- Decreased production (bone marrow not making enough)
- Ineffective production (not working right)
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What are the causes of marrow insufficiency?
- Depression
- Depletion
- Destruction
- Degeneration
- Lack of hormonal input
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What is marrow depression?
Marrow temporarily loses ability to make segs in response to peripheral demand (see a decreas in segs, but no bands)
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What is marrow depletion?
- High peripheral demand (no more left)
- Degenerative left shift results
- Improves in time if animal lives
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What is marrow destruction?
- Blood-forming cells destroyed
- Decrease in all cell types (red and white)
- Due to chemical/physical agents (estrogen, chloramphenicol, radiation) - bone marrow doesn't come back
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What causes a lack of hormonal input? What are the different hormones and where are they releases)
- Chemical messenger that give marrow the message aren't producing the hormone
- Erythropoietin (renal)
- Leukopoietin (released by dying WBC)
- Thrombopoietin (on platelet surface)
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Where do we do WBC estimations?
Buffy coat or on a smear (with lots of practice)
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What the normal WBC reference ranges for dogs/cats and horses/ruminants?
- Dog & Cats: 5,000 to 20,000 cells/ul
- Horses & Ruminant: 5,000 to 12,000 cells/ul
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When do you do a corrected WBC count?
If you see any NRBCs on a smear, you must do a corrected WBC count
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What is a NRBC?
Nucleated RBC
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What is the formula for a corrected WBC count?
(100/100 + NRBCs) X WBC count
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What do you look for when doing a WBC differential?
- Size and shape of nucleus
- Chromatin pattern
- Nuclear:Cytoplasmic ratio
- Color of cytoplasm
- Any cytoplasmic granules
- Any cytoplasmic vacuoles
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Which is the most numerous granulocyte?
Neutrophils
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Where do granulocytes function?
Function in the tissues and are only transported in blood
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Whats the technical name for a neutrophil?
Polymorphoneutrophil
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How big are neutrophils?
12 - 15 microns in diameter
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What does the cytoplasm of a neutrophil look like?
Pale (light pink/slightly purple with neutral granules)
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What is a Dohle Body?
- Blue dot in cytoplasm - 1 per cell
- Means there is inflammation
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What do eosinophils look like?
Look similar to neutrophils but granules are pink/orange/red
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What are the reasons for an increase in eosinophils?
- Parasitic infections (internal or external)
- Allergic reactions
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What do basophils look like?
Granules are basophilic
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What can basophils be easily confused with?
- Darkly stained eosinophils
- Mast cells
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Where do we find mast cells?
Should not be in blood - should only be in tissue
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Why are immature granulocytes occastionally found in blood?
Due to large tissue demand (bone marrow can't keep up so sends them out early - bands) or due to granulocytic leukemia
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Describe lymphocytes
- Smaller than neutrophils
- Larger than RBCs
- Stain blue/purple
- High nuclear:cytoplasmic ratio
- Small and large lymphs are common
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What is the most common WBC in ruminants and rodents?
Lymphocytes
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What is the most common WBC in dogs and cats?
Dogs and Cats
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How long do lymphocytes live?
3 days to years
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What does the nucleus of monocytes look like?
oval or kidney bean shape
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What do macrophages look like?
- Large cell in tissue
- Abundant pale blue cytoplasm
- Vacuoles
- Pale vesicular nucleus
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How many more neutrophils are stored in bone marrow as they are in peripheral blood?
30 times more
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How many neutrophils are marginated (stored) along capillary walls and do not normally circulate?
Half
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What will we see in a stree leukogram?
- Neutrophilia without a left shift
- +/- monocytosis
- Eosinopenia
- Lymphopenia
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What kind of cell is this?
Band Neutrophil (no segments)
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What kind of cell is this?
Basophil (basophilic granules)
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Describe the chromatin pattern of this neutrophil.
Clumpy & Hypersegmented
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What is in the cytoplasm?
granules
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What are the clear spots in the cytoplasm?
Vacuoles
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Describe the chromatin pattern of this neutrophil.
Diffused
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What is the blue spot on this neutrophil?
Dohle Body (one blue granule in segment cytoplasm)
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What is the spot on this neutrophil?
Ehrlichia Canis (intracellular parasite)
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What kind of leukocyte is this?
Eosinophil (eosinophilic granules)
-
What kind of leukocyte is this?
Eosinophil (eosinophilic granules)
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What are these spots in the cytoplasm of these neutrophils?
Histoplasma Organisms (fungal)
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- A: Segment
- B: Platelet
- C: Lymphocyte
- D: Eosinophil
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- A: Lymphocyte
- B: Segment
- C: Eosinophil
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What kind of leukocyte is this?
Lymphocyte (stain blue/purple, high nuclear:cytoplasmic ratio)
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What kind of cell is this?
Mast Cell (many dark purple granules, may obstruct view of nucleus)
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What kind of cytoplasmic basophilia does this basophil have?
mild to moderate
-
What kind of cytoplasmic basophilia does this basophil have?
moderate to severe
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What kind of leukocyte is this?
Monocyte (vesicular (lacey) chromatin, cytoplasm gray-blue, vacuoles in cytoplasm)
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What kind of leukocyte is this?
Neutrophil (segment)
-
What is wrong with these neutrophils?
Pelger-Huet Anomaly
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What kind of cell is this?
Plasma cell (eccentric nucleus with perinuclear clear zone)
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Describe this neutrophil.
S shaped band
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What is in the cytoplasm of this neutrophil?
Viral inclusion body
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