Purpose of making this presentation is to provoke knowledge of veterinary science towards modern world

1. Bone Marrow Examination
In All Domestic Animals
Dr.Sama Awais Akram
DVM
College Of Veterinary And Animal
Sciences Jhang

2. Bone Marrow Examination
• Bone marrow examination refers to the
pathologic analysis of sample of bone marrow
obtained by bone marrow aspiration and
bone marrow biopsy (often called Trephine
biopsy)

3. • The aspirate yields semi-liquid bone
marrow, which can be examined by
• under a light microscope
• analyzed by flow cytometry
• chromosome analysis
• polymerase chain reaction (PCR).
• Frequently, a trephine biopsy is also
obtained, which yields a narrow,
cylindrically shaped solid piece of bone
marrow which is examined microscopically
for cellularity and infiltrative processes.

4. Materials Required
Needles (Salah needle, Kalima needle,
Jamshidi needle)
Anti-coagulant (EDTA)
Microscope
Slide and Cover Slip
Stains
(Leishman’s stain, Giemsa’s stain, PAS stain,
Wright’s stain, Peroxide stain, Prussian blue
Stain)

5. .

6. Collection of bone marrow Sample
• By two methods
1-Aspiration
2-Core Cutting/Biopsy

7. .
Collection of sample by Aspiration

8. Indication for Bone Marrow Aspirate
• To establish or conform the diagnosis of Leukemia
• To determine, if deficiency of one or more blood type in
the peripheral blood is due to deficiency of precursor in
bone marrow
• To determine the cause, if possible, of refractory
anemia especially if interference with erythrocyte
maturation is suspected (Vit.B12 and Folic acid
deficiency, lead poisoning).
• To demonstration of metastatic neoplasm
• To demonstration of certain type of infection
(histoplasmosis, toxoplasmosis)
• To demonstrate deficiency in iron store (iron deficiency
amenia)

9. Aspiration Sites
Dog: Iliac crest preferred, sternum & ribs also.
Cat: Proximal end of Femur through trochanteric fossa
Birds: Sternal ridge (keel) or tibio-tarsus.
Small Ruminants: iliac crest of hip bone & ribs.
Large Ruminants: Femur, Humerus, Sternum & Ribs.
Calf: Vertebral spines good site.
Horse: iliac crest, tuber coxea, sternum, ribs.

10. Preparation of Smear
1- Before it clots, eject the aspirate on slanted slide & make
smear.
2- Removal excess blood by capillary tube improve quality of
bone marrow smear.
3- Smear can be made immediately by fresh marrow.
4- Aspirate place in EDTA Vacutainer
5- Strains commonly used
>Wright’s Strain
>Prussian blue reaction---- For iron granules
>Peroxidase --- To diff. granulocytes from lymphocytes

11. Look for marrow fragment while
making smear

12. Scheme for examining BM aspirate
At Low Power (10X)
>Determine cellularity
>Identify Megakeryocytes
>Look for Clump of Abnormal cells
>Identify Macrophage

13. .
• At high Power (40X,100X)
>Identify all Stages of maturation of M:E cells
>Determine the M:E ratio
>Perform a Diff. Count
>Look for areas of BM necrosis
>Assess the iron content

14. .
Interpretation of BM Examination

15. Cells of the bone marrow
• Erythroid series
• Myeloid series
• Megakaryocytic series
• Monocytic series

16. Erythroid series
Proerythroblast
Early erythroblast (Basophillic )
Intermediate erythroblast (Polychromatic)
Late erythroblast (Orthochromatic )
Myeloid series
Myeloblast promyelocyte myelocyte  metamyelocyte  band form  mature
neutrophil

17. Megakaryocytic series
Megakaryoblasts
Promegakaryocytes
Megakaryocytes
Monocytic series
Monoblast
Promonocyte
Monocyte

19. .Cellularity
1- If bone marrow particles on avg., <25% adipose tissue,
bone marrow is Hypercellular.
2- If………………………………………………., >75% ............
…………………….,……………Hypocellular.
Megakaryocyte
1- If more megakaryocytes are present then megakaryocytic
hyperplasia.
2- If less megakaryocytes are present then megakaryocytic
hypoplasia.
3- The presence of immature form of megakaryocytic series
indicate increased megakaryocyticpoiesis, usually
megakaryocytic hyperplasia.

20. Compare BM
Hypocellular Hypercellular Normo

21. Arrow show Megakaryocytes

22. .M:E (G:E) ratio
G:E ratio 1:1 means both cells are equal.
1-if G:E ratio increases means granulocyte
hyperplasia or erythrocyte hypoplasia so
animal is anemic.
2-if G:E ratio decreases means granulocyte
hypoplasia or erythrocyte hyperplasia.

23. Average of M:E ratio
Species Avg. M:E ratio
Dog 1.25
Cat 1.63
Horse 0.93
Cattle 0.71
Goat 0.63
Sheep 0.50

24. .Macrophage
Relative %age of macrophage is determined.
Incease in macrophage indicate certain
pathological processes.
Iron Content
Iron pigment in hemosiderin will range from
yellow to yellow-green to yellow-brown.
Complete assessment obtain by iron stains
> Prussian blue or Mallory stain

25. Stored Iron Stain Blue

26. .
Evaluation of Diseases Associated with
BM Examination

27. A: Hematologic Diseases
Deficiency of cell from any cell lines
Non-regenerative or Non-responding Anemia
Excessive production of Neutrophils or lymphocytes
Hematopoietic and non-hematopoietic
Presence of immature or morphologically atypical
cells in circulation
Erythrocytosis ,espacially splenic contraction or
Cardio-pulmonary disease
Thrombocytopinea

28. B: Systemic Diseases
Congenital hematologic disorders
Hematolymphoid neoplasms: i.e leukemia, chronic
myeloproliferative disorder, myeloma, etc.
Osteomyelitis
Iron deficiency
Specific diseases: histoplasmosis, lishmaniasis,
myeloma, storage diseases.
Hyperproteinemia
Hypercalcemia

29. Reference
1- OUTLINE OF Veterinary Clinical Pathology
BY ( MAXINE M. BENJAMIN )
2- VETERINARY CLINICAL PATHOLOGY 1ST EDITION
BY ( AHRAR KHAN )
3- CLINICAL PATOLOGY 2ND EDITION
BY ( Sabitri Sanyal . Aparna Bhattacharyya )
4- Fundamentals of Veterinary Clinical Pathology
BY ( Stockham Scott )

30. .
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