GETTING MOST OUT OF HEMATOLOGY IN

CATS & DOGS

What is your Minimum Data Base?
Hematology – Engine Light
What makes HEMATOLOGY test GOLD STANDARD ?

❑ Numbers

❑ Graphs

❑ Microscopy
5 Golden Rules for hematology testing
1. Garbage in – Garbage out (Sample quality)

2. Reticulocytes are part of hematology

3. You need 5 part differential

4. You have to look at the graphs

5. Use your microscope no longer than 2 mins.

1. Garbage in - Garbage out (Sample Quality)
Sample collection and management
2. Reticulocytes are part of hematology
CBC – Data
Erythrocytes

Red Blood Cell mass — RBC, HCT, HGB

Erythrocytes
Red Blood Cell mass — RBC, HCT, HGB

1/3 of RBC is HGB

HGB X 3 = HCT

• If it is greater than this it could be caused by hemolysis

 Erythrocytes
Erythrocytes indices — MCV, MCH, MCHC, RDW

• MCV ( Mean corpuscular volume ) : Volume of the cells

• MCH ( Mean corpuscular hemoglobin) : Quantity of hemoglobin per

avarage RBC

• MCHC (Mean corpuscular hemoglobin concentration ) : Cellular hemoglobin

concentration per avarage RBC – Apprx 1/3 of Hct if RBCs are in normal size!

• RDW : Variation in RBC size

Classifying Anemia – Reticulocytes

Dog : %1 in health

Cat : % 0.4 (Aggregates retics)

Anemias are common

• 203.939 animals were tested for CBC in 3 months and 18. 975’ of them ( % 9.3 ) had anemia

2DeNicola DB, Matthews JA, Ferandis PJ, Frye MB. Comparison of reticulocyte counts to mean corpuscular volume
and mean corpuscular hemoglobin in anemic dogs. Paper presented at XIIth Congress of the International Society of
Animal Clinical Biochemistry; May 22–26, 2006; Istanbul, Turkey.
Reticulocytes –IDEXX Announcement

• Helping you to classify anemia: regenerative or not

• For non-anaemic patients, it will help you to detect underlying disease
such as occult haemorrhage or haemolytic disease.

Talking about the previous run and looking at trends PERSISTENT increase!

For example, dogs with occult haemorrhage following GIT ulceration due to
NSAIDs may have a reticulocytosis !

Canine: 10 to 110 K/μL Feline: 3 to 50 K/μL

Kayla’s haematology
Kayla’s hematology

Reticulocytosis

normal
Kayla’s blood film
Schistocytes
Kayla’s splenic haemangiosarcoma
Factors affecting MCV

 MCV
• Reticulocytosis
• FeLV infected cats
• Poodle breed
• Erythrocyte agglutination
(false)
 MCV
• Iron deficiency
• Porto -systemic shunts
• Akita breed
Factors affecting MCHC

 MCHC  MCHC
• Always a result of in vivo or in • Iron deficiency
vitro hemolysis or lipemia
• Increased amount of Hb can
not be produced in the cell
3. Increase confidence with 5 part differantial
Leukocytes
WBCs : Granulocytes / Agranulocytes

Neutrophils Lymphocytes Monocytes Eosinophils Basophils

6 part differential
How many dogs with severe
inflammatory leukograms
have normal or low WBC ?

50- 60 %
WBC data are used to answer the following questions :
1. Is there evidence of inflammation?

2. Is there evidence of a glucocorticoid (stress) or epinephrine (excitement) response?

3. Is there a demand for phagocytosis or evidence of tissue necrosis?

4. If inflammation is present can it be further classified chronic?

5. Is there evidence of hypersensitivity or allergic reaction?

Inflammation Leukogram (Acute)

✓ Neutrophilia + LEFT SHIFT ( Band Neutrophil)

✓ Lymphopenia
✓ Mild monocytosis (Demand for phagocytosis)
✓ Eosinopenia or Eosinophilia (systemic allergic or hypersensitivity reaction)
Stress Leukogram
(Glucocorticoid leukogram)

✓ Primary changes lymphopenia (750 – 1500/µl)

✓ Eosinopenia second most common changes
✓ Mild neutrophilia
✓ Mild monocytosis but this is not a consistent change
Stress Leukogram

LEM (O) N
Schnookers - Hematology

3 part- diff????

Inflammatory Leukogram with

Superimposed Strees Leukogram
Raleigh- Hemogram

3 part- diff?
Epinephrine Leukogram

✓ Mild neutrophilia – dogs

✓ Lymphocytosis and neutrophilia – cats
✓ This type of response dissapates in minutes
Demand for phagocytosis or
Evidence of tissue necrosis ?

✓ Monocytosis indicates a demand for phagocytosis or tissue necrosis

✓ Monocytosis is almost always present in chronic inflammatory condition,

but it can also occur with acute inflammation

✓ Monocytosis > 4000/ µl : Prepare blood film to look for phagocytosed agents
( opsonized RBCs in IMHA , E. canis, histoplasma vs.)
If inflammation is present, can it be further
classified as chronic?
Classic acute inflammation

WBC count is elevated due to the granulocytes

✓ Neutrophilia + LEFT SHIFT (Band Neutrophil) – Regenerative Left Shift

✓ Lymphopenia
✓ Mild monocytosis (Demand for phagocytosis)
✓ Eosinopenia or Eosinophilia (systemic allergic or hypersensitivity reaction)
Classic chronic inflammation

Most cases WBC is normal or slight elevation!

✓ LEFT SHIFT DISSIPATES ! (normal or slightly elevated neutrophil )

✓ MONOCYTE is almost always elevated

✓ Normal or elevated lymphocyte

✓ Mild Non – regenerative anemia ( PCV %30 – dog / % 25 cat)

✓Normocytic- normochromic anemia (Normal MCV & MCHC)

Evidence of “hypersensitivity”

Persistent eosinophilia and/or basophilia

• Parasitic diseases with systemic component (heartworm, flea bite dermatitis)

• Feline astma

• Allergic gastroenteritis

• Eosinophilic granuloma complex in cats

4. The importance of graphs…

RBC
RBC

WBC WBC
Optical
Fluorescence
Technology
Impedance Technology
2. Impedance Technology
NUMBER of cells

Example from an in clinic

analyser

RBCs

Platelets

SIZE of cells
NUMBER of cells

RBCs

Platelets

SIZE of cells
Cats have relatively small RBCs and more
variety in the size of their platelets
NUMBER of cells

Cat
RBCs

Cat
platelets
SIZE of cells
Grouping the cells only by the size is not enough
NUMBER of cells

Feline
RBCs

Feline
Platelets

SIZE of cells
On every haematology report from an impedance analyser, the user
must look at the histogram and ask ‘is the cut in the right place?’

So they may provide a magnification

The impedance analyser tries the same things with the white blood cells:
classify by size alone
NUMBER of cells

Example from the VetScan HM5

SIZE of cells
Examples from the field
Examples from the field
Grouping the cells by size alone is inherently limited
Hematology Analyzers
Procyte Dx

Laser flow cytometry

Optical fluorescence

Laminar flow impedance

CBC: RBC-PLT Dot Plot

Red blood cells

Size (Forward Scatter)

Reticulocytes

Platelets DNA + RNA

Side Fluorescence
A Tale Of 2 Dot- Plots

Josie Exley

Which one is regenerative?

Cutie, 2 years old , FS, Greyhound

• 4-week history of lethargy and depression

• Febrile 4 weeks ago

• Lost some weight

• Pale mucous membranes

Josie
Cutie -ProCyteDx
IRON is needed to make hemoglobin
MAKING TATA MAKING JAGUAR
Cutie-Leukogram
Cutie- Differantial Diagnosis
• GI Bleeding
Cutie is 2 year old
• tumor and doesn’t have fleas…
• ulcer
• hookworms Look for
• External bleeding hookworms!
• tumor
• fleas
Leukocytes
Granularity (complexity)

Neutrophil

Eosinophil
Fluorescence
CBC: WBC Dot Plot

Side Fluorescence

Granularity (Side Scatter)

RETICULOCYTE MATURE RBC

BAND MATURE
NEUTROPHIL NEUTROPHIL
Blizzard - Leukogram

Band neutrophil – blue cytoplasm

with significant amounts of RNA

Mature neutrophil – pale blue cytoplasm

with minimal amounts of RNA
Shark fin!
Montana

Which cell populations are interfering with each other?

Case study- Sassy

Sassy Patient
Sassy is a 5 year old FS Long haired Dachshund

Presenting Complaints
• Acute onset of weakness and collapse
• Fever

History
• Current on vaccines and parasite prophylaxis
Sassy
Sassy - Erythrogram
Sassy - Erythrogram
Reticulocytosis in
patient compared
to normal

Increased number
of small cells or
RBC fragments
Sassy - Leukogram
Sassy - Hematology
Saline Slide Agglutination Test
Autoagglutination
Sassy - Leukogram

• Leukocytosis
• Neutrophilia with suspected
left shift
• Lymphocytosis, Monocytosis
• nRBC presence
Sassy - Leukogram

nRBCs
Compliance tips-the “broken car part”!
Questions