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Hematology basics pt 1 jpg
1. THE BLOODY TRUTH
ABOUT CATS AND DOGS
HEMATOLOGY BASICS: DIFFERENTIALS
Sarah Ouellette, CVT
2. Why we run a CBC
• Provides broad overview of overall health status
• Used for:
• General health
• Sick patients
• Pre-anesthetic
• Recheck abnormalities
3. Hematology Basics
• Collection/handling
SAMPLE QUALITY IS A MAJOR CONTRIBUTOR
TO ANALYTICAL ERRORS
• Poor collection/handling methods can lead to:
• Inadequate cell counts
• Morphologic artifacts
4. Hematology Basics
• EDTA
• anticoagulant of choice
• LTT
• Correct ratio
• Low blood volume can cause
• False increase in plasma protein
• False decrease in PCV/RBC count
• Morphologic artifacts
5. Things to remember!
• Make blood smears soon after collection
to reduce the risk of artifacts
• Especially when sending out for pathology
review
• Make a good quality smear
• unfixed
6. Why we do a differential
• CBC analyzers do NOT give us any information
about specific morphological changes or specific
cell types
• CBC analyzers only “suspect” abnormalities such
as nRBC/band cells, etc.
8. Blood smear
• Staining:
• Wright’s stain
• New methylene blue
• IVG protocol:
• 10 dips in each stain or 30 seconds/stain
9. Blood Smear Staining
• Why its important? Overstaining
• Not reliable for accurate results
10. Reading a blood smear
• NEVER NEVER NEVER go looking for
something abnormal – unless indicated
• Examples:
• Automated analyzer suspects an abnormality:
• Nucleated RBC
• Band cells
• Abnormal (high/low) cell count
12. Erythrocytes
• Non-nucleated cells
• Biconcave disk
• Prominent central pallor
• Stain pink/salmon color
• Function: transport oxygen between lungs
and tissues
• Hemoglobin is the O2 carrying component of the cell
13. Erythrocytes
• Canine:
• Lifespan = 110-120 days
• Slightly larger than cats
• Mild polychromasia
• Abnormalities that can be seen in healthy dogs:
• Occasional nucleated RBC
• Occasional howell jolly bodies
14. Erythrocytes
• Feline:
• Lifespan: 65-76 days
• Smaller than canine RBC’s
• More variable in size
• Little to no central pallor
• Abnormalities that can be seen in healthy cats:
• Occasional heinz bodies
• Small # of howell jolly bodies
15. Abnormal Hemogram
• Polychromasia
• Color variance with RBC
• Indicative of immature RBC within the blood
• MILD variance is accepted in cats and dogs
• Causes:
• Anemia
• Bone marrow disease
**importance of staining**
16. Abnormal Hemogram
• Hypochromasia:
• Increased central pallor
• Cells stain less overall
• Low hemoglobin content
• Causes:
• Iron insufficiency
• Chronic blood loss
• Anemia
17. Abnormal Hemogram
• Poikilocytes or poikilocytosis:
• General term used to describe RBC’s with an
abnormal shape
• Some abnormalities have specific significance
relating to a particular disease while others may
be non-specific
18. Poikilocytes
• Acanthocytes (spur cells)
• 2-10 blunt/club shaped projections
• Different lengths
• Irregular intervals
• Causes:
• Dogs with hemangiosarcoma
• Increased blood cholesterol, iron deficiency
anemia, renal disease, and occasionally some liver
diseases
19. Poikilocytes
• Echinocytes (spiculated or crenated cells)
• Numerous sharp/blunt projections
• Same length
• Evenly spaced around the cell
• Causes:
• Incidental
• excessive EDTA:Blood ratio causes dehydration of cells
• Drugs
• salicylates, phenylbotazone, Lasix, and certain
chemotherapeutic agents
20. Ecchinocytes
• Shown at right are two
smears from the same
sample, the upper one
made while the blood was
fresh
• The lower made after
overnight storage of blood
at refrigerator temperature
21. Poikilocytes
• Eccentrocytes
• Ragged fringe of cytoplasm along one side of cell
• Dense staining
• May rupture (hemi-ghosts or “pseudo-spherocytes”)
• Causes:
• Oxidative damage to cell
• Ex. onion toxicity
22. Poikilocytes
• Keratocytes (helmet cells)
• Blister-like defect along the perimeter
• May rupture (bite-shaped)
• Causes:
• DIC
• Vasculitis
• Hemangiosarcoma
• Iron deficiency anemia
• Hepatic lipidosis in cats
26. Poikilocytes
• Stomatocytes
• Elongated mouth-like area of central pallor
• Due to the absence of hemoglobin
• Causes:
• Regenerative anemias
• Liver disease
• Lead poisoning
• Non-clinical: blood smear that is too thick
29. Poikilocytes
• Codocytes (target cells)
• Cluster of hemoglobinized cytoplasm within the
area of normal central pallor
• Only observed in dogs
• Causes:
• Cholesterol/phospholipid abnormalities
• Regenerative anemia
• Liver disease
• Non-clinical - Excessive amounts of EDTA
30. Target cells
• Upper panel:
numerous target
cells in the blood of
a dog with
cholestasis.
• Lower panel: target
cells in a dog with
regenerative
anemia
31. RBC Inclusions
• Red blood cell inclusions can arise from a
variety of sources but is important in
providing insights into
metabolic, physiologic and pathogenic
conditions
• Inclusion = nuclear or cytoplasmic aggregates (proteins)
present within the cell
32. RBC inclusions
• Howell-jolly bodies
• Small round basophilic inclusion
• Deeply staining non-functioning nuclear remnant
• Rare in normal dogs
• Cats may have low #’s
• Removed by the spleen
• Causes:
• Regenerative anemias
• Splenic disease
34. RBC Inclusions
• Distemper
• Aggregates of viral particles
• Found in RBC,WBC, epithelial cells
• Rarely seen even with positive infection
• Larger than Howell-jolly bodies
35. Abnormal Hemogram
• Basophilic stippling
• Spontaneous aggregation of ribosomal RNA in the
cytoplasm of young erythrocytes
• Causes:
• Regenerative anemia
• Bone marrow disorders
• Lead poisoning (absence of anemia)
36. RBC inclusions
• Heinz Bodies
• Small pink/clear short blunt projection
• Precipitates of denatured or oxidized hemoglobin
• Uncommon in dog/ common in cats
• Confirm w/ New methylene blue stain
• Causes
• Onion toxicity (dogs)
• Endogenous HB in cats
• (diabetes, lymphoma, hyperthyroidism)
37. Heinz bodies
• Blood from a cat with
Heinz body hemolytic
anemia associated with
acetaminophen toxicity.
Upper panel: the large
Heinz bodies are causing
severe distortion of the cell
outline.
Lower panel: stained with
New Methylene Blue.
38. Abnormal Hemogram
• Metarubricytes (nucleated RBC)
• Late stage nucleated red blood cells
• Chromatin is markedly clumped and the cytoplasm is
more abundant than that of a lymphocyte
• Causes:
• Regenerative anemias
• Acute lead toxicity
• Bone marrow disease
• Septicemia
39. Metarubricytes/Nucleated RBC
• Left panel: metarubricyte from a dog.
• Middle panel: basophilic rubricyte from a dog.
• Right panel: a megaloblastoid nRBC from a cat with
myelodysplastic syndrome.
40. Abnormal hemogram
• Rouleaux formation
• Arranged in overlapping chains
• Stacks of coins
• Common in cats
• Weak clumping (proteins)
• Usually not significant
42. Abnormal hemogram
• Reticulocytes
• Young non-nucleated cells
• Retained ribosomal DNA
• Appear as small dark blue dots within the cell
• Polychromatophils stained with New Methylene
blue stain
• Causes:
• Anemia
• Bone marrow disease
44. RBC Parasites
• There are several organisms that can be
detected from examination of a peripheral
blood smear.
• Some organisms can cause significant
disease in the host while others can be
non-pathogenic.
45. RBC parasites
• Babesia canis
• Protozoal parasite transmitted via brown dog tick
• Hemolytic anemia seen in in mild dz
• Intravascular hemolysis seen in severe dz
• B. canis appear usually as a single or multiple pear
shaped organisms within the cell
• Babesia in endemic in greyhounds.
46. RBC parasites
• Mycoplasma haemofelis
• Formally known as hemobartonella
• Transmitted through infected fleas
• Epicellular bacterial parasite
• Observed as small blue cocci, rings, or rods on
the edges or the surface of the red cells
• Can be mistaken for stain participate
• Causes hemolytic anemia
47. RBC parasites
• Mycoplasma haemocanis
• Formally known as hemobartonella
• Transmitted through infected fleas
• Epicellular bacterial parasite
• Recognized as chains of cocci across the face
of the red cell
• Can be mistaken for stain participate
• Rarely causes anemia in dogs
48. Artifacts
• Artifacts can significantly impair the
examination of a blood smear
However,
They can be easily avoided
• The most common cause of these artifacts is
error in handling, collection, storage and
preparation of the blood smear
50. Artifacts
• Water
• Caused by precipitation in fixative
• Moth eaten appearance
• Refractile
• As you focus up and down on
the microscope the artifact
“flashes”
** Always want to make sure stains
covers are on tight**
Same sample first stained with IVG protocol and the second stained for a 1-2 walked away and came back
One should never go looking for an abnormality, if you ware specifically looking for something you are more likely to “imagine” or overidentify an abnormality than identify a real abnormality
Hemoglobin = the carrying component of oxygen
Occasional nucleated RBC and howell jolly bodies can be seen in healthy patients and are considered normalMild polychromasia (color variance) can also be seen and is normal
Causes: the body is trying to kick out more cells from the marrow to compensate for a loss however the cells are not fully matured yet
In dogs they have an increased zone of central pallor with a thin rim of cytoplasm and in cats a visible pallor is seen. The red blood cells as a whole stain very pale compared to normal cells. Picture: Blood film from a dog with chronic blood loss resulting in iron deficiency anemia.Note that, in addition to the hypochromic cells (with increased areas of central pallor), a variety of other shape abnormalities are present (keratocytes, schistocytes).
Poi.kilo.cy.tosisNow we are going to talk about the different kinds of RBC shapes and what they meanCause: liver or splenic diseases, bone marrow disease, or iron deficiency
Most commonly Dogs with hemageosarcomacells can also be seen in dogs with increased cholesterol, iron deficiency anemia, renal disease, DIC, and occasionally some patients liver disease.
eKIN.O.sitesAlso referred to as spiculated red blood cells, however crenated or crenation of cells is most commonly used. With this abnormality numerous sharp or blunt projections are seen evenly spaced around the circumference of the cell. This is the most common morphologic artifact in blood smears. Drugs: not common but can be seen
E.centro.sitesThe thin membrane of the RBC is removed or ruptures and leaves behind a fragmented cell that lacks a central pallor.This type of abnormality is seen in oxidant injuries in dogs and catsOxidative means it damages or affects the hemoglobin in the RBC on a molecular level making it ineffectivePicture:Three eccentrocytes from a dog with onion toxicity
This is a blister like defect in the red blood cell perimeter commonly called helmet cells. Disseminated intravascular coagulation (DIC) is a serious disorder in which the proteins that control blood clotting become over activeThese cells are sometimes seen in cats but have no clinical significance. When present in increased numbers or viewed along with other red blood cell abnormalities it is considered a significant finding.
However cats with liver disease such as hepatic lipidosis or a portosystemic shunt can have these cells present along with other acanthocytes.IMAGES: Blood smears from cats with hepatic lipidosis. Upper panel: Non-anemic patient with relatively few altered red cells.
Indicated by red arrows
oval/rectangular slit or mouth-like clearing in the center of the cell due to the absence of hemoglobin
Pyruvate kinase deficiency is an inherited lack of the enzyme pyruvate kinase, which is used by red blood cells. Without this enzyme, red blood cells break down too easily, resulting in low levels of these cells (hemolytic anemia
Canine IMHA: nearly all the cells in this field are spherocytes, as indicated by their decreased diameter, dense staining properties, and lack of central pallor
Cholestasis is any condition in which the flow of bile from the liver is slowed or blocked
Nucleated red blood cells and polychromasia are commonly accompanied with basophilic stippling.
Common in cats – ineffective spleens, their cells are more susceptible to oxidant injury'sPathologic in dogs but not in catsEndogenous - Having an internal cause or originHas to do with oxidative damage to cells
Many stages of development, the late stage is most commonly seen
Normal stain they are called Polychromatophils and appear bluer than mature red blood cells
A definitive diagnosis is based upon finding the organism in a peripheral blood smear and/or a positive immune fluorescent assay (IFA) result. Endemic = of a disease or condition) Regularly found among particular people or in a certain area
Bacteria present from stain… found others not on RBC in smear when I looked around.
