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Specimens should be labeled with (4-7)
- date sample was taken, species, ID, initials
- optionally, age, history and symptoms of specimen
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Preferred way of drawing blood sample
vaccutainer bolder with needle attachment or as large a needle as possible, gently put into tube
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Hemolysis
- breaking apart or destruction of RBCs caused by trauma. Pink or red serum.
- Can effect chemistry and hemology results.
- can be caused by alcohol
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Hemolysis-affected results.
- diluted sample leads to falsely lower concentrations (PVC, RBC count, WBC count)
- higher levels of certain constituents (K+, ALP, enzymes, LDH, Ca, Mg, LD, AST, ALT, GGT, folate, iron.
- interferes with lipase and bilirubin determinations
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Blood for CBC
anticoagulated. Lavender top, EDTA tube.
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CBC consists of (9)
- RBC count
- WBC count
- platelet count
- PVC or hematocrit
- hemoglobin
- blood smear (differential, automated, give counts, morphology, platelet estimate, inclusion bodies and blood parasites)
- Total plasma protein
- erythrocyte indices (MVC, MCH, MCHC)
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erythrocyte indices
MCV, MCH, MCHC
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reticulocyte count
- performed to check for degree/type of anemia
- COunt of reticulocytes (must be anticoagulated blood)
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EDTA tube
- purple top, used for CBCs. Anticoagulant.
- Spun, plasma only used for cTnI (cardiac troponin I)
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Heparin tube
Green top. Can be used for CBC, not great for WBC. Used in exotics, plasma can be used for chemistries, so good when only a small amount of blood available.
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Sodium citrate tube
- blue top. Not used for CBCs.
- Clotting factor tests.
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Sodium floride tube
grey top. Rare in vet. Used in glucose tolerance tests
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Red stopper tube
nothing inside. Coated with silicone to prevent sticking. Yields serum after spinning.
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Red/tan striped stopper tube
(tiger top). Serum separator. Contains wax plug between serum and clot. Yields serum, you can pour serum out without a pipette.
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supernatant
removable liquid part of a sample after centrifugation.
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Plasma
liquid part (supernatant) of anticoagulated blood
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serum
liquid part (supernatant) of clotted blood.
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PCV
packed cell volume. Made up of RBC, buffy coat and plasma.
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Total protein
told with plasma on a refractometer (scales for proteins and specific gravity)
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Estimated platelet count
- Count platelets in 10 oil immersion fields on stained blood smear, multiply by 15,000. Platelets per microliter.
- Look at feather edge for clots.
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Why do corrected WBC count?
Automated counter counts nRBCs as WBCs, have to subtract for accuracy.
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Hypersegmented neutrophils
five lobes or more. Can be from EDTA or old neutrophils, or disease/inherited condition
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band cell
immature neutrophil.
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metamyelocyte
VERY early neutrophil, marked left shift, usually seen with many band cells. Cytoplasm darker.
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Myelocyte
Extremely immature neutrophil, younger than metamyelocyte, very rarely seen.
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Lymphocyte
- Can be small or large. Small has less cytoplasm and is smaller. Cytoplasm is pale blue.
- a-typical, abnormal, reactive (granulation)
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Toxic changes in neutrophils
- signifies rapid reproduction, can be too much EDTA or old blood too.
- Least to most severe is dohle bodies, cytoplasmic basophilia, vacuoles or foaminess, intensely stained primary granules (toxic granulation), change in nuclear size and shape (or change in nuclear cytoplasmic ratio).
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Red blood cell changes
basophilic staining (common in mature ruminants, sometimes in cats with anemia, lead poisoning in dogs)
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Heinz bodies
- oxidative injury to RBCs. 5-10% normal in cats.
- Nipple-like projection of denatured hemoglobin
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Howell-Jolly bodies
nuclear remnants, stain blue. More significant in dogs than cats (more common in cats)
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Precursors to platelets
megakaryocytes
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Where we look on a blood slide
everywhere, then monolayer
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crenation
pointed projections around cell, caused by smear not being dry before fixing.
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rouleaux
stacking (coins), normal in horse.
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Agglutination
clumping of cells, immune mediated. Can try saline agglutination test
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polychromasia
blue tint to cell (usually macrocytic, reticulocytes)
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hypochromia
decreased staining in cell due to decreased hemoglobin.
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stomatocytes
rod shaped pallor across cell. no significance unless there are lots, then inherited.
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anisocytosis
variation in size of cell. Microcytosis or macrocytosis.
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poikilocytosis
change in shape, general term (must be qualified with specifics)
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acanthocytes
spur cells. uneven blunt projections. Hepatic lipidosis, hermangiosarcoma, dogs with liver diease
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echinocyte
burr cells. Spiculated cells with evenly spaced and sized projections. Not blunt.
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Spherocytes
round, smaller than normal, darkened cells with no/little central pallor. IMHA
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Leptocytes
thin, folded or distorted cells. Target cells. Too much EDTA or precursers to acanthocytes
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schistocytes
fragmented cells.
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keratocyte
incompletely fragmented cells
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eccentrocytes
displacement of hemoglobin because opposing sides of the RBC membranes have fused due to oxidative injury.
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anulocytes
bowl shaped erythrocytes, too much membrane flexibility
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Dacrocytes
teardrop shaped erythrocytes seen in myelofibrosis. Artifact if all tails point same directions
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pencil cells
oval elliptical cells or elongated.
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ghost cells
intervascular trauma
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Babesia
large teardrop shaped intracellular organisms, seen at feather edge. Looks like a vacuole. Tick-bourne.
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Hemobartonella felis
short dark purple rods on periphery of cell. Also mycoplasma haemofelis.
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Ehrlichia canis
intracellular parasite in monocytes and neutrophils. Looks like small clusters called morulae.
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Refractile droplets within RBCs. Which stain is messed up?
Fixative has water in it.
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Plasma fibrinogen
spin 2 microhematocrit tubes. Read plasma protein on one, put the other in 56 degree C water for 5 minutes, then read again. Fibrinogen has precipitated.
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Coagulation tests
usually blue top. Whole blood clotting time, activated clotting time, bleeding time
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Ways to get RBC and WBC in birds and reptiles
Natt and Herrick's solution or eosinophil method (indirect)
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How to count on a slide (manual)
backward "S"
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What do you need to calculate an absolute count?
Differential and absolute count (percentage vs. absolute number)
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Four things in a SNAP test
lyme, heartworm, ehrlichia, anaplasmosis
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Mice and rats
- granulocyte nuclei are doughnut shaped.
- Basophils may be confused with mast cells (appear with cardiocentesis)
- Hamsters, gerbils and chinchillas have same hematological features
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Guinea pig hemology
larger RBC than other rodents. Kurloff bodies.
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Rabbit hemology
- neutrophils have eosiniphilic granules and are called heterophils (used to be called pseudo-eosinophils). Same function.
- Eosinophils have larger granules.
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Ferret hemology
Inhalant anesthetics decrease RBC and hematocrit, as much as 33%.
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Llama hemology
thin, elliptical erythrocytes. Gives higher oxygen carrying capacity due to high altitude
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Colors of plasma
- red: hemolysis (slight v. marked)
- yellow: icteric (normal in horse)
- white and cloudy: lipemic
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Blood stains
- 1. Fixative
- 2. eosin stain for RBC, eosinophils and some parts of parasites
- 3. Wrights-Giemsa or hematoxylin stain for nuclei of WBC, inclusion bodies and parts of parasites
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Order of most frequently found WBC in cats and dogs
- No Lazy Monkeys Eat Bananas
- neutrophils, leukocytes, monocytes, eosinophils, basophils
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nRBC
metarubrocyte. immature RBC (before reticulocyte)
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barr body
female sex indicator, little nodule off neutrophil nucleus
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dohle bodies
toxic change on a neutrophil, small light grey staining area in cytoplasm of neutrophil
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How much of an atypical cell do you report?
write down all, but 5% significant
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lymphoblast
HUGE lymphocyte
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Plasma cell
looks like a lymph with more cytoplasm, peripheral nucleus, vacuoles called russell bodies
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distemper inclusion body
large greyish (bluish?) body in lymphocyte.
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Largest RBC in rodents is in
guinea pigs
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kurloff bodies
HUGE inclusion, pushing nucleus of lymph to side. Sometimes in guinea pigs
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Dif between basophilic rubricyte and thrombocyte in bird
Basophilic rubrocyte is bigger, has dark cytoplasm. Throbocyte has very pale cytoplasm.
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Small lymphocyte vs thrombocyte in bird
Small lymph is bigger
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Blood parasite plasmodium common name
malaria. Avian plasmodium.
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leukocytozoan
extracellular, big leaf-shape. Exotic
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hemoproteus
big darker vacuole inside of cell. Exotic
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Azurophil
in reptiles, like monocyte. Same function.
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If you cannot positively identify an inclusion you classify it as....
an inclusion. Don't classify unless you can tell.
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