Lab 5 Flashcards
Normal Hemoglobin measurement?
18-20 mol/l
Oxygen binding capacity of Hemoglobin is increased by?
- Decreased 2,3 DPG level in RBCs.
- Decreased pCO2 level in the blood (ex. in case of respiratory alkalosis)
- Decreased temperature of blood (hypothermia)
- Increased pH of the blood.
Oxygen binding capacity of Hemoglobin is decreased by?
- Increased 2,3 DPG level in RBCs.
- Increased pCO2 level in the blood (ex. in case of respiratory acidosis)
- Increased temperature of blood (hyperthermia)
- Decreased pH of the blood (acidosis, respiratory or metabolic).
What is oxygen saturation (SAT%)?
The percentage of oxygenated Hemoglobin molecules compared to the whole amount of Hemoglobin molecules in one unit blood.
Normal values of SAT%?
Arterial blood: 95-99%
Venous blood: 80-90%
Methemoglobin?
Hemoglobin molecules containing oxidized iron (3+ form).
Causes of increased hemoglobin concentration?
- Usually associated with different types of relative (dehydration)
- Absolute polycythemia.
Causes of decreased hemoglobin concentration?
- Usually associated with relative (hyperhydration)
- Absolute oligocythemia (anaemia).
Normal RBC count?
4.5-8 x 10^12/IT/l
In order to calculate the indices we must measure?
Ht or PCV (hematocrit, packed cell volume), red blood cell count, hemoglobin concentration.
Normal Mean Corpuscular Hemoglobin (MCH) value?
12-30pg
In young animals it (and MCV) can be increased 28-32pg
Decreased Mean Corpuscular Hemoglobin (MCH)?
Hypochromasia
Increased Mean Corpuscular Hemoglobin (MCH)?
Hyperchromasia
Mean Corpuscular Volume (MCV) indicates the average size of the RBCs, which is?
- Increased: Macrocytic
- Normal: Normocytic
- Decreased: Microcytic
Normal Mean Corpuscular Volume (MCV) value?
60-70fl
Causes of microcytosis?
- Chronic blood loss
- Iron, copper, pyridoxine (vitamin B6) deficiency.
- Portosystemic shunt
Causes of macrocytosis?
(mostly regenerative anaemia)
- Polycythemia absolute vera (erythroleukemia)
- Vitamin B12, folic acid, cobalt deficiency.
- Erythroleukemias
Mean Corpuscular Hemoglobin Concentration (MCHC) indicates?
The average concentration of hemoglobin in erythrocytes (Hb concentration)
Normal Mean Corpuscular Hemoglobin Concentration (MCHC)?
300-350g/l (30-35%) - Normochrom
Decreased Mean Corpuscular Hemoglobin Concentration (MCHC) - hypochromasia?
- Newborn animals
- Regenerative anaemias
- Iron deficiency anamias
Increased Mean Corpuscular Hemoglobin Concentration (MCHC) - hyperchromasia?
- Erythroleukemia (polycythemia absolute vera)
- Vitamin B12, folic acid, cobalt deficiency
- Immunhemolytic anaemia (spherocytosis)
- Lead poisoning
- Splenectomy
Typical changes in deviated parameters?
- Macrocytic, hypochromic
- Noromocytic, normochromic
- Microcytic, hypochromic
- Microcytic, normochromic
- Macrocytic, normochromic
Macrocytic, hypochromic: Increased MCV, decreased MCHC, (increased reticulocytes)?
Regenerative anaemias
Noromocytic, normochromic: Normal MCV, normal MCHC, normal or decreased MCH?
Non regenerative anaemias
Microcytic, hypochromic: Decreased MCV, decreased MCHC, (decreased Hb synthesis)?
Iron, copper, pyridoxine deficiency anaemias, liver failure, portosystemic shunt.
Microcytic, normochromic: decreased MCV, normal MCHC?
Japanese Akita (normal)
Macrocytic, normochromic: increased MCV, normal MCHC, impaired DNA synthesis?
FeLV infection, vitamin B12, Co or folic acid deficiency, erythroleukemia, poodle macrocytosis
Normal Red Cell Distribution (RDW) values?
Dog: 12-16%
Cat: 14-18%
Normal Platelet Distribution Width (PDW) values?
Dog: 6-8%
Cat: 7-12%
Normal Corrected Reticulocyte Percentage (CRP) values?
<1-2% (without anaemia)
Cause of increased reticulocyte count?
Different types of regenerative anaemia:
- Acute blood loss (approx. 3-5 days are needed for the bone marrow to increase the reticulocyte count in the blood).
- Haemolytic anaemia
- Chronic blood loss
- Nutrient deficiency anaemias.
What is osmotic resistance of RBC dependent on?
- The pH of plasma
- The reagents
- Temperature
- Osmotic concentration of plasma
- Reagents (NaCl concentration)
- RBC membrane status
- Regenerative status (reticulocytes are more resistant)
- HbF (fetal hemoglobin) content of the RBCs (fetal RBCs, containing HbF are more resistant)
Proper staining methods?
May-Grünwald staining, Giemsa staining, Diff Quick staining
Check gross signs?
- Rouleau formation: coin arrangement (horse often, dog, cat, swine sometimes, cattle rare)
- RBC aggregates
- Large cells (horse often)
- Thrombocyte aggregates
Intensity of staining of RBCs?
- Polychromasia, hyperchromosia: more intensive staining
- Hypochromasia: Weak staining
Polychromasia, hyperchromasia?
More intensive staining
- RNA, or nuclear remnants,
- more Hb -regenerative process
Hypochromasia?
Weak staining
- Decreased Hb-content
- or other nutrient deficiency
Size of RBCs?
- Macrocytosis: many big cells
- Microcytosis: many small cells
- Anisocytosis: variable cell size - iron deficiency and regenerative process
- Poikylocytosis: variable size and color
RBC types?
- Young and nucleated RBCs (in order of malnutrition):
#Proerythroblast
#Basophil erythroblast (normocyte, normoblast)
#Polychromatophil eryhtoblast (normocyte, normoblast)
#Acidophil erythroblast (normocyte, normoblast) - Young but mature RBC without nucleus:
#Reticulocyte - Appearance of young RBCs:
#Increased production (regenerative anaemia), spleen
or bone marrow disease, leukemia, extra medullar
erythrocyte production, Pb toxicosis (with basophil
punctates), hyperadrenocorticism.
Reticulocyte - appearance?
Increased production (regenerative anaemia) - chronic Fe deficiency anaemia, hemolysis, acute blood loss, chronic blood loss.
Inclusion bodies in RBCs?
- Heinz body
- Howell-Jolly body
- Basophilic punctuates
- Hb inclusions
Heinz body - appearance?
Heinz body (NMB - new methylene blue stain): denatured HgB
Appearance:
- O2 effect,
- oxidative damage to RBCs (cat!, ex. methaemoglobinaemia),
- GSH deficiency
Howell-Jolly body - appearance?
Nuclear membrane remnants. Appearance: - Vitamin B12 deficiency, - increased production of red cells, - splenectomy
Basophilic punctuates - appearance?
Nuclear remnants Appearance: - Regenerative process - Young RBCs of cat - Physiological in ruminants - Lead poisoning
Hb inclusions - appearance?
- Hb damage
- Increased RBC production
- Regenerative anaemia
RBC parasites?
- Haemobartonella canis, felis, bovis
- Babesia spp. (canis, gibsoni), B. canis is very common in
Hungary - Ehrlichia canis, Equipment etc.
- Dirofilaria immitis, repens
- Anaplasma marginale, centrale, ovis
- Eperythrozoon wenyoni, ovis, suis, parvum
- Citauxzoon felis
- Theileria parva, mutans, annulata, hirci, ovis
- Trypanosoma cruzi, congolense, vivas, rucei, evans, suis,
equiperdum - Leishmania donovani
Normal serum iron (SeFe) value?
18-20 umol/l
Causes of low serum iron concentration?
- Chronic blood loss
- Decreased intake (piglets, calves)
- Impaired gastric, duodenal, jejunal function (reduction,
transport, absorption)
Causes of high serum iron concentration?
- Iron toxicosis (overload)
Total iron binding capacity (TIBC)?
50-68 umol/l
Causes of low Total iron binding capacity (TIBC)?
- Chronic inflammation (negative acute phase protein)
- Chronic liver failure (decreased transferring synthesis in
the liver) - Neoplastic disease
Causes of high Total iron binding capacity (TIBC)?
- Iron deficiency anaemia (not severe: normal iron level + high TIBC, severe: low iron level + high TIBC)
Laboratory findings in hemolysis?
- Decreased PCV
- Decreased haptoglobin
- Decreased RBC osmotic resistance
- Increased reticulocytes (regenerative anaemia)
- Increased total bilirubin
- Increased indirect bilirubin
- Increased lactate dehydrogenase (LDH) I, II.
- Increased urobilinogen and Hgb in urine
- Polychromasia, poikilocytosis
- Leukocytosis, (neutrophilis)
- Spherocytosis
- Jaundice
- Hyperchromic stool
How do we measure haemoglobin concentration of the blood?
By using Spectrophotometric method (Drabkin-method). Or by calculating: PCV (l/l)/3 * 1000 = HGB (g/l)
What is included in the measured haemoglobin result?
It is a sum of Hgb molecules from the haemolysed RBCs and the very small amount of free Hgb content of the plasma, which is usually bound to a carrier protein (haptoglobin).
What is haptoglobin?
A carrier protein of free Hgb content of plasma.
Is there an increase of haemoglobin concentration in blood in case of intravascular haemolysis?
There is NO notable increase in Hgb concentration in case of intravascular hemolysis.
What does the oxygen dissociation curve describe and what can shifts to the left or right show?
Its a curve that plots the proportion of hemoglobin in its saturated (oxygen-laden) form on the vertical axis against the prevailing oxygen tension on the horizontal axis.
Left shift: Decreased temp., decreased 2-3 DPG, decreased H+.
Right shift: (reduced affinity), increased temp., increased 2-3 DPG, increased H+.
What is methaemoglobin? What is the mechanism of methaemoglobin and can it carry oxygen?
Hgb molecules containing oxidized form (3+ form) are called methaemoglobin.
These are unable to carry oxygen.
Can methaemoglobin be converted to normal haemoglobin?
Yes. They are reduced to normal hemoglobin by methaemoglobin-reductase enzyme.
Which animal has a lower haemoglobin concentration in younger animals?
Swine. Young pigs have much lower Hgb concentrations than older ones.
Does the haemoglobin affinity for oxygen change with age?
Usually not, except pigs.
What method(s) are used to count red blood cells?
Bürker-chamber method, automatic cell counter, or calculations: (Ht l/l)/(5) x 100 = RBC count x 10^12/l.
How do the sizes of red blood cells of young animals, the Japanese Akita and Poodles differ?
Japanese Akita have small RBCs (55-65 fl).
Poodles have very large RBC (75-80 fl).
What is red cell distribution width and platelet distribution width?
Give a number that is correlated with the range of the average size of the RBCs and platelets.
What do a large and a short RDW suggest?
Short RDW: non regenerative processes.
Large RDW: regenerative process.
How do we perform a reticulocyte count?
Staining: Brilliant-cresil stain.
Counting: Count 100-1000 RBCs and take the percent of the reticulocytes.
What animals do not have reticulocytes in their plasma and where do they have them?
No reticulocytes appear in horses or ruminants, they appear only in the bone marrow, not in the peripheral blood.
Which animals commonly have more punctuated forms of reticulocytes?
Cats.
What are the differences between nucleated and non-nucleated reticulocytes?
Nucleated RBCs are too young to carry oxygen.
What is the normal reticulocyte count?
2-3%
Why would a reticulocyte count or percentage sometimes need to be corrected?
Because mature RBCs are more sensitive to any damage than young RBCs and reticulocytes, therefore in case of RBC damage, usually more mature RBCs are died than young ones.
What is the normal corrected reticulocyte count (CRC) value?
<0,06 x 10^12/l (without anaemia)
What can damage red blood cell membranes?
- Nephropathy (uremia)
- Specific membrane damage (immunohemolytic anaemia)
- Increased physical damage (Ex. long-distance running)
What other lab measures could be taken in connection with RBCs?
- Serum iron measurement
- Total iron binding capacity (TIBC)
What is and how do we measure the serum iron concentration?
Serum samples are needed for this analysis, because fibrinogen content of the plasma may disturb the measurement.
Fe3+ is reduced to Fe2+ by ascorbic acid.
Fe2+ reacts with ferrosin and forms a red colored chelate (complex molecule) which can be measured photometrically.
What should serum iron measurement always be performed together with?
TIBC (Total iron binding capacity) analysis
In which species can we use reticulocyte count?
All except horses and ruminants?
What are the reticulocytes?
Young, dont have nucleus or granules.
What is MCH?
Mean corpuscular hemoglobin indicates average Hb content of RBCs.
What is MCHC?
Mean corpuscular hemoglobin concentration indicates the average concentration of hemoglobin in erythrocytes .
