Leukocyte identification and evaluation

Question 1

Leukocyte identification

Answer 1

Identification of white blood cells found in the peripheral blood
A leukogram- is the quantification of the different classes of leukocytes as well as ny morphologic abnormalities of the cells
Calculated by evaluating a peripheral blood smear
Species differences

Question 2

How to examine a blood smear

Answer 2

Start by examining th slide on the 40x objective
-Examine the slide for platelet clumps, enlarged cells and microfilariae
Go to the feathered edge of the slide
-Look for evidence of platelet or cell clumping
Move 3-4 fields inwards to the monolayer
-This is where you will find approx. 200 cells/field
-Cells are single layer→ they are not overlapping and they are not touching
Go to 100x objective
-Perform the differential cell count
-100 WBC will be counted- this will give you a percentage for each cell type observed

Question 3

Absolute values

Answer 3

Once your differential cell count is performed and your total WBC count is known, absolute values of the WBCs can be calculated

Question 4

Neutrophils are

Answer 4

Also known as “segs”
Nucleus: irregular, elongated and lobed

Question 5

Segmented neutrophils

Answer 5

Mature neutrophil- the most abundant leukocyte in peripheral blood in small animals
Granulocytic cell

Question 6

What do neutrophils do

Answer 6

Participate in the inflammatory response
Increased numbers generally indicate inflammation of infection
Phagocytosis of foreign invaders or disease self

Question 7

what do neutrophils look like

Answer 7

Small, very poorly stained granules
species/individual differences: colourless to lightly staining granules
Cattle- granules stain faintly pink, giving cytoplasm an orange-pink tint overall
Horses- fine diffuse granules
May contain small vacuoles, cells will be larger if they are actively phagocytic
Larger cells may have a bubbly or foamy appearance
Reptile, birds, some fish and some mammals (rabbits and guinea pigs) have heterophils in the place of neutrophils
Granules are eosinophilic

Question 8

Band neutrophils are and look like

Answer 8

Immature stage of neutrophils
Has a smooth, parallel sided, horseshoe-shaped nucleus with “clubbed ends” and no constrictions, the chromatin is visible
If the narrowest part of the nucleus is greater than ⅔ the width of the widest part of the nucleus, the cell is classified as a band neutrophil
When unsure of what maturity stage a cell is, always classify it as the more mature stage
Normal values for all species: 0-3%

Question 9

Eosinophils are and function

Answer 9

Granulocytic cell
Nucleus similar to neutrophils
Primary function is in immune system modulation
Also capable of phagocytosis
Numbers increase with allergic reaction and parasitic infections
Found in small numbers in peripheral blood in healthy animals

Question 10

What do eosinophils look look

Answer 10

Contain granules that are large and stain red
They vary considerably b/w species
Canine: granules are light reddish colour and will range from small to large within the same cell, 2-10%
Feline: granules are rod-shaped, small, numerous and stain a reddish colour, 2-12%
Equine: bright orange-red granules are very large, often filling the cell and occluding the nucleus, 0-4%
Bovine: granules are relatively small, round and red, 2-20%

Question 11

Basophils are and functions

Answer 11

Granulocytic cell
Involved in the deviation of immune system
Will see increased numbers with inflammation or infection
Not often found in peripheral blood smears

Question 12

Basophils look like

Answer 12

They are larger than neutrophils
Have nucleus similar to monocytes
They have granules that vary among species
Canine: small number of dark- purple-blue/black granules
Feline: large, round, lavender granules
Equine/Bovine: large number of dark blue/black granules that may contain the nucleus (0-3%

Question 13

Lymphocytes are and function

Answer 13

Agranulocytic cell
Function is the humoral immune response (fever, etc) and cell mediated responses (antibody production)

Question 14

Lymphocytes look like

Answer 14

Dark purple- blue round to oval or slightly indented nucleus (kidney bean shape)
The nucleus will have variations of its colour due to the clumping of chromatin
Small amount of clear to sky blue cytoplasm, may appear to have none
Most abundant cell in the blood smear of ruminants
Variation in lymphocyte size based on species
Cats and dogs have smaller lymphocytes with scant cytoplasm
Cattle have medium to large (neutrophil size) lymphocytes with more abundant cytoplasm
Cytoplasm may have pink/purple granules

Question 15

What does an increase of lymphocytes indicate

Answer 15

Viral infection

Question 16

Monocytes are and functions

Answer 16

Classified as agranulocytes cells
The largest of the WBCs
Participate in the inflammatory response
Numbers are often increased with chronic infections
Capable of migrating into tissues
They phagocytize foreign material and dead, dying or diseased cells
Responsible for normal erythrocyte destruction and associated metabolic iron recycling
The most misidentified cell on blood smears

Question 17

Monocyte looks like

Answer 17

Nucleus varies in shape, but is generally kidney shaped with occasional lobes
A soft cell, so it can fold over on itself- giving it weird shapes
Nucleus does not stain as dark as other cells and appears to have a lacy appearance
Cytoplasm is grey-blue
The cytoplasm may contain small, pink granules or vacuoles

Question 18

Monocyte identification

Answer 18

Cytoplasm is grey-blue with ground glass appearance, sometimes tiny pink granules
Nucleus is usually deeply indented and may show amoeboid characteristics
Large and small vacuoles may be preset
Chromatin (in the nucleus) is reticulated (lacy)

Question 19

Lymphocyte identification

Answer 19

Cytoplasm is true or sky-blue no granules except the occasional large one
Nucleus is round to slightly indented (occasional deep indentation)
Vacuoles are extremely rare and are tiny if present
Chromatin is clumped

Question 20

2 systems used to identify abnormal leukocyte morphology

Answer 20

Slight, moderate, marked
Slight: ~10% of cells affected
Moderate :~25% of cells affected
Marked: >50% of cells affected
1+,2+,3+,4+
1+: 5-10% of cells affected
2+: 10-25% of cells affected
3+: ~50% of cells affected
4+: >75% of cells affected

Question 21

Abnormal leukocyte morphology

Answer 21

Nuclear hypersegmentation
Toxic change
Intracytoplasmic inclusions
Atypical and reactive lymphocytes
Smudge cells/basket cells
Neoplasia

Question 22

Nuclear hypersegmentation is

Answer 22

Canine or feline neutrophils with more than 5 lobes
Usually due to ageing of cells either in the patient or in the blood tube
Commonly seen

Question 23

Toxic change of leukocytes is

Answer 23

The most common disease-induced cytoplasmic change in neutrophils
Found in association with inflammation, infection and drug toxicity
If severe, it is often in association with a bacterial infection
Most significant when found in dogs
Common in cats that are not severely ill
Cytoplasmic basophilia

Question 24

What can be seen in leukocytes with toxic change

Answer 24

Cytoplasmic basophilia
Dohle bodies
Blue staining cytoplasmic inclusions
Can be single or multiples
Vacuoles or “foamy” appearance to the cytoplasm
Toxic granulation (rare)
Intensely stained granules
May be significantly larger than normal neutrophils

Question 25

How to quantify percentage of neutrophils with toxic change

Answer 25

Mild 5-10%
Moderate 11-30%
Marked >30%

Question 26

Severity of toxic change within the cell

Answer 26

Dohle bodies 1+
Mildly basophilic 1+
Moderately basophilic with dohle bodies 2+
Moderately basophilic and foamy 3+
Basophilic with toxic granules 3+

Question 27

Intracytoplasmic inclusions can be caused by

Answer 27

Canine distemper inclusions
RBCs and neutrophils
Ehrlichia and anaplasma
Others including
Histoplasma capsulatum
Francisella philomiragia
Mycobacterium
Hepatozoon canis
Leishmania infantum

Question 28

Reactive lymphocytes are and look like

Answer 28

Basophilic cytoplasm
More abundant cytoplasm
Increased cell size
Larger and more convoluted nucleus
Also called immunocytes
Seen with antigen stimulation
This will occur with antigen exposure through infection of vaccination

Question 29

Atypical lymphocytes look like

Answer 29

Basophilic cytoplasm
Increased cell size
Cleaved nuclei
Asynchronous maturation of the nucleus and the cytoplasm
Azurophilic granules
Chronic antigen stimulation
Esp. Canine Ehrlichiosis
May be present in normal bovine lymphocytes

Question 30

Smudge/basket cells are

Answer 30

Ruptured, degenerative leukocytes
Significant when large numbers are present- leukaemia
Can be an artefact

Question 31

Neoplasia and blood cells is caused by and looks like

Answer 31

Neoplastic proliferation of hematopoietic cells
Also known as leukaemia- there are neoplastic cells in the blood and/or bone marrow
Neoplastic blood cells in the peripheral blood are also known as type cells
They include
Blast cells, atypical cells, bizarre cells, mitotic figures, unclassified neoplastic cells- nucleus appears to belong to one cell line, while the cytoplasm appears to belong to another cell line

Question 32

Mitotic figures are indicative of

Answer 32

when seen on the blood smear are indicative of neoplasia

Question 33

Blast cells are

Answer 33

confined to the bone marrow in relatively small numbers
With neoplasia, blast cell numbers will increase, or they will be present in the blood smear

Question 34

Abnormal lymphocytes with neoplasia look like

Answer 34

will be vacuolated, clumped together or appear more basophilic
They are usually seen at the feather edge of a smear and are rare

Question 35

When to consider neoplasia

Answer 35

Neoplasia must be considered if you are seeing bizarre cells or if an unexpected cell type predominates on the blood smear

Question 36

Why does neutrophils change consideration for neoplasia

Answer 36

The only exception being that of the neutrophil series
Toxic change is common with significant inflammation
Immature neutrophils may be present in peripheral blood with severe infections
The clinical history must be considered

Question 37

Myeloproliferative disease is seen where and in what animals

Answer 37

Characterized by bone marrow
That has hypercellularity
A loss of orderliness in maturation
Abnormal (neoplastic) cells
Not uncommon in cats
Often infected with FeLV
Characterised by the presence of abnormal RBCs, WBCs or platelets
More than one cell line may be affected

Question 38

CBC may see what with myeloproliferative disease

Answer 38

Increased WBC
Increased nucleated RBC count
Decreased PCV
Abnormal cells in the blood smear
Nonregeneratice anemia
Thrombocytopenia

Question 39

Lymphoproliferative disorders are used to descibe

Answer 39

Can be used to describe any abnormal proliferation of lymphoid cells
More commonly used to describe tumors that are derived from lymphocytes or plasma cells
More common than myeloproliferative disorders

Question 40

Lymphoproliferative disorders are common in

Answer 40

More common in cats
Often associated with FeLV or FIV
Also seen in cattle with bovine leukosis virus (BLV)

Question 41

Lymphoproliferative disorders are characterized as and can be detected in what test

Answer 41

Categorized as primary lymphoid leukaemia, lymphoma, or plasma cell tumors
We can use PCR testing to distinguish b/w neoplastic and non-neoplastic lymphoproliferative disorders

Question 42

The main difference between lymphoid leukemia and lymphoma is

Answer 42

the distribution of disease
Lymphoid leukemia- neoplastic cells are in the bone marrow +/- blood
Lymphoma- neoplastic cells are in solid tissue in the form of tumors (ex. Lymph nodes)