hematology lab

how many hours should dog be fasted before blood collection

6-8 hrs

2 hr for water

what should be avoided before blood collection

excerize/ play

stress

hemolysis interference with blood parameters

hemolysis free hemoglobin

mild hemolysis free hemoglobin

100mg/dl

moderate hemolysis free hemoglobin

200

severe hemolysis free hemoglobin

>400

when dose primitive hematopoiesis take place

in visceral yolk sac (extraembryonic mesoderm)

dog breeds asso w macrocytosis

Miniature and Toy Poodles. 

dog breed asso w microcytosis w/o anemia

Akita, Shiba Inu, and Chow Chow, and potentially the Siberian Husky

dog breed asso w stomatocytosis

Alaskan Malamutes, Drentse Patrijshonds, and Miniature Schnauzers

when dose definitive hematopoiesis take place

takes place in AGM region of embryo

what was a good Cv system important for

activity in warmer waters and in air conditions => also the hemopoetic system (blood-forming system)

conserved pattern (similar traits) found in

vertebrates (high and lower)

what was key to survival of stem cells

different potencies (mutli-toti-puli) key to survival

what helped remodell chromatin

conserved structure of H1 in chromosomes (a key part in genetic material)

HSC molecular diagnostics

CD 34 ag- used to identify and isolate hematopoietic stem cells (HSC)

GATA 1 gene- transcription factors essential for development of RBC, plts and mast cells

CD 34 ag

used to identify and isolate hematopoietic stem cells (HSC)

GATA 1 gene

transcription factors essential for development of RBC, plts and mast cells

how can CD34+ cells from 3 week old be done

using canine anti-CD34 mAB

after female dog with XSCID (Canine X-linked Severe Combined Immunodeficiency) received CD34+ male cells they showed improved

lymphocyte counts and T-cell and B-cell response (immune resp) after 2 months

how was male origin of cells confirmed

male donor T cells contained SRY gene

what is critical for normal RBC and PLT formation

GATA-1

GATA-1 mutant (changes) effect on erythroid precursor

GATA-1 mutants (a mutation/ change) have blocked committed erythroid precursors maturation and undergo aptosis (RBC precursors cant mature and die)

megakaryocytes (cells that become plts) stay immature and proliferate

if case with laboratory marrow and blood results above (RBC aptosis and megakaryocytes immature and proliferating

GATA1 mutation should be preformed

what is erythropoeisis influenced by

EPO

GATA1

feline metarubriyte, MGG staining

metarubriyte- aka nucleated red blood cell or NRBC

are reticulocytes larger or smaller than mature erythrocytes

larger = described as macrocytes

what regualtes erythropoisis

EPO

EPO

• growth factor

• glycoporteins (165 aa)

where is EPO (erythropoietin) produ

produ by fibroblasts of kidney in adult mammals

terminolgy

rubriblast

proerythroblast

prorubricyte

basophilic erythroblast

basophilic rubricyte

basophilic erythroblast

polychromatophilic rubricyte

polychromatophilic erythroblast

metarubricyte

orthochromatic erythroblast

feline blood smear

• erythro smaller

• varible in size (ansiocytosis)

canine blood smear

• larger than feline

• more uniform in size

• more pronounced central pallor

equine blood smear

• largest of both

• distinct central depression rather than clear cental pallor

polychromasia

RBCs exhibit a bluish-gray tint when stained with specific dyes, indicating an increased number of immature red blood cells (reticulocytes).

This is often a sign of the bone marrow responding to anemia or other conditions by releasing these immature cells prematurely into the bloodstream. 

hypochromasia

red blood cells that have less color than normal under a microscope. This usually indicates a reduced amount of hemoglobin, the protein responsible for carrying oxygen in the blood, within the red blood cells. A common cause of hypochromia is iron deficiency. 

echinocytes

indicate:

• kidney failure- uremia

• liver diease- prob w lipid metab'

acantocytes

schistocytes

• fragmented

• various shapes

• iregular/ spikey edges

indicate:

• microangiopathc hemolytic anemia (MAHA)

• TTP

• HUS

• DIC

microcytosis

codocytes

• have central, round dark area of hemoglobin

• pale ring

• outside of pale ring there is darker ring

  => look like target

indicate

• liver disease

• post haemorr regen

rouleax

rouleax

stacking of RBC

what animals have circulating reticulocytes in health

dogs and cats

what animals have circulating reticulocytes during regenerative response only

cow

sheep

laboratory markers for diag of anemia

HCT

RBC

HGB

severity of anemia

level of HCT in dog in moderate anaemia

20-29%

level of HCT in dog in severe anaemia

<19%

level of HCT in cats in moderate anaemia

14-19%

level of HCT in cats in severe anaemia

<13%

indication body is responding to anemia

high reticulocyte (young RBC) count => suggesting regeneratuve response

non regen reticulocyte count in anemia

normocytic nomochromic cells in blood smear

bone maroow aspirate evaluation- test to asses bone marrows ability to regenerate blood cells

in cattle what reflects regneration

nucleated RBC

in ru what do howell jolly bodies suggest

regenerative anemia

what can cause howel jolly bodies and NRBC

• glucocorticoud treatment

• prolonged storage of EDTA blood

mild anemia in equine

30-33%

moderate anemia in quine

20-29%

severe anemia equine

<19%

in horses what suggests response to anemia

macrocytosis

ansiocytosis

cause of regen anemia

1. haemorrhage

2. hematolysis (hemolytic anemia)

TP conc during haemorrhage

decrease

TP conc during hematolysis- the breakdown or destruction of red blood cells

increase in TP

cause of hematolysis

1. immune mediated autoagglutination- coombs test

2. heinz bodies

3. young age/ worms

heinz bodies

clumps of damaged hemoglobin in RBC

aggregates of oxidised, precipitated hemoglobin

strong indicator of IMHA

spherocytosis

coombs test

• RBC w IgG and C3 bound to cell membrane

• incubation w ab anti-C3 and anti-IgG (coombs reagent)

purpose of coombs test

detect antibodies in surface of RBC which can indicate immune mediated hemolysis (Immune-mediated hemolytic anemia)

ab present in positive commbs test

IgM and IgG

heinz bodies

aggregates of oxidized, precipitated hemoglobin

monocyte

left shift results in increase in…

bands (more immature cells such as metamyelocytes and myelocytes)

degenerative left shift

more band neutrophils are counted than segmented neutophils

acute leukemia

• cytopenias, blast (immature blood cells) present

• more that ~25 of blast- AML or ALL

chronic leukemias

• cytophilia of involved cell types

• cell hyperplasia- CML or CLL

diag tool for leukemia

• blood smea

• bone marow

• immunophenotyping/ cytochemistry

• PARR

• flow cytometry

markers if AmMegL (acute megakaryroblastic leukemia)- subtupe of AML

mAB anti CD61 and myeloperoxidase

blastic cells- immature cells

stained feline blood smear

SNAP tetst

an example of monoclonal ab use

what could steroid cause

neutrophilia

deccrease in lymphocyte count (lymphopenia)

granularity

reactive lymphocyte

• increase in number in response to infections or inflammation

• help fight infection

parasites in blood

white intracellular EHrlichia canis infection

babesia canis

babesia canis

• pear/ teardrop shape

what are piroplasms

parasitic protozoa

transmitted by ticks and infect RBCs

cause babesia

ex.

babesia

Cytauxzoonosis felis

Cytauxzoonosis felis

(more clear image can see black dots which are parasite)

bacteria in blood- haemobartonella felis

• appearing as small, blue cocci, rings, or rods, typically on the edges or surfaces of red blood cells

microfilaria- blood worms

change in PLT count

• thrombocytosis

• thrombocytopenia

clinical signs of thrmobocytopenia

• petechiation

• ecchymosis (bruising)

• hematuria

Na- EDTA

dependent pseudothrombocytoenia

• plts clump together → EDTA = atrifically low plt count, hence pseudo,

in blood colection what is added to APTT

kaolin

cephalin

to inittiate intinsic pathway coagu cascade

in blood colection what is added to PT

thromboplastin

to initate extrinsic pathwway of caogu cascade

incubation at

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