LECTURE: BLOOD #1

Kavita R. Lall, B.Sc. (Hons.), D.V.M. (Hons.), M.Sc. (Dist.)

 Learning objectives

 Introduction

 Erythrocytes

 RBC metabolism

 Fate of RBC

 Abnormalities of RBC concentration

 Understand what blood is, including its components

 Discuss the different properties and functions of blood

 Explain the commonly used method to determine blood’s specific

gravity

 Differentiate between plasma and serum

 Discuss erythrocytes and how the shape differs in different species

 Provide an overview of erythropoiesis

 Describe the metabolism and fate of RBCs

 Describe the two types of erythrocyte destruction

 Appreciate the lifespan of erythrocytes in different species

 Discuss abnormalities of RBC concentration

 Blood: Fluid connective tissue that flows throughout the body in the blood
vessels of the cardiovascular system

 Plasma - fluid portion (55-70% TBV) Total blood Volume

 Water: 91-92%
 Solids: 8-9% - proteins and organic and inorganic compounds
 Yellow to colourless depending on the quantity, species and animal’s diet

 Suspended cellular elements (30-45% TBV)

 Erythrocytes
 Leukocytes
 Thrombocytes
 is a measure of the yellow colour of serum. This colour is
 Icterus index (plasma or serum) normally due almost exclusively to the presence of bilirubin, a
hemoglobin waste product from the red blood cells. The icterus
index is expressed as a number of “plus” signs (from zero to
++++). An icterus index of zero is normal.
 Estimates the total bilirubin concentration

 It is measured by comparing the colour of the plasma with

a standard solution of potassium dichromate K2Cr2O7
  The normal pH of the blood is ~ 7.4
 Dog: 7.32 - 7.68
 Cattle: 7.35 - 7.50
 Horse: 7.35 - 7.43
 Fowl: 7.56

 Venous blood: Slightly more acidic due to increased CO2

Venous blood is deoxygenated blood which travels from the peripheral blood vessels, through the venous
system into the right atrium of the heart.
The venous system refers to your network of veins and the way your veins connect with other
blood vessels and organs throughout your body.
 Acidosis and alkalosis
Acidosis and alkalosis are conditions in which there is a disturbance in the pH balance (acid-base balance)
of the body. Acidosis is a condition in which the bodily fluids become too acidic, with an abnormally low pH
level. In alkalosis, the opposite is true: the fluids of the body are too alkaline (high in pH).
 Functions of blood

Transportation Regulation/maintain Protection

homeostasis

Oxygen and carbon pH/acid base balance Provides immunity

dioxide
Nutrients Body temperature
Hormones Water balance

Waste products Osmotic pressure

 Specific gravity of blood

 Specific gravity - the ratio of weight of a given volume of a fluid

to the weight of the same volume of distilled water measured at
25°C

 Plasma protein concentration is largely responsible for

the specific gravity of the plasma

 The cellular elements have higher specific gravity than

the plasma
 Copper sulphate method: Commonly used manual method of
hemoglobin determination based on specific gravity

 When a drop of blood from a patient with normal hemoglobin

values is dropped into a copper sulfate solution, it falls rapidly to
the bottom; if the drop falls slowly or not at all, hemoglobin
levels are below reference range
  Various factors influence the specific gravity of the blood

 Specific gravity is increased:

 Polycythemia: High altitude, newborns

Polycythemia, also called erythrocytosis, refers to increased red blood cell mass, noted on laboratory evaluation as
increased hemoglobin and hematocrit levels.
 Severe dehydration: Excessive fluid loss such as in vomiting,
diarrhoea

 Hemoconcentration: Loss of plasma, e.g. burns

An increase in the concentration of blood cells resulting from the loss of plasma or water from the blood stream.
 Hemodilution helps maintain a proper volume of blood without
transfusion. In this technique, some of the patient's blood is
 Specific gravity is decreased: removed shortly before surgery and replaced with a water- and
mineral-based solution (see volume expander), diluting the
blood.
 Pregnancy: Increase in plasma volume (hemodilution)

 Anaemia: Reduced RBCs

 Renal diseases: Loss of albumin Albumin is a protein made by your liver. Albumin enters your bloodstream
and helps keep fluid from leaking out of your blood vessels into other
tissues.
 Hemodilution: Hypersecretion or prolonged treatment with glucocorticoids
Excess glucocorticoids, either from hypersecretion or prolonged treatment, can cause water retention and fluid shifts that dilute the blood, resulting in a
decrease in blood specific gravity. This is due to increased plasma volume, reduced ADH activity, and electrolyte imbalances that impact the concentration of
cells and solutes in the blood
 Starvation and malnutrition: Decrease in plasma protein
Plasma protein concentration is largely responsible for the specific gravity of the plasma
 Intravenous fluid transfusions
IV fluid transfusions decrease blood specific gravity by increasing plasma volume and diluting the concentration of blood components such as red blood cells, proteins, and
electrolytes. This occurs because the added fluids increase the ratio of water to solutes in the bloodstream, reducing the blood's overall concentration and specific gravity.
 Viscosity of blood

 Viscosity is influenced by the concentration of RBCs and plasma

proteins

 Among plasma proteins, the viscosity is mainly contributed by

the gamma globulins which provide the resistance to blood
flow and helps the normal pumping activity of the heart

 It is normally about five times greater than water

 Viscosity of blood is determined using a viscometer

 An increase in the viscosity of blood is seen in conditions such

as polycythemia, congestive heart failure, jaundice, vomiting,
diarrhea

 A decrease in the viscosity is commonly noticed in anemia,

oedema Swelling in the ankles, feet and legs is often caused by a build-up
of fluid in these areas, called oedema
 Serum

 Fluid that remains from plasma after clotting factors have been
removed by clot formation
The serum is the liquid or undiluted part of the blood, which lacks clotting factors. Hence, it is formed after
blood coagulation. The serum helps in the diagnosis of diseases such as blood pressure, sugar,
cholesterol, etc.
 It differs from the plasma in lacking fibrinogen, prothrombin
and other coagulation factors which are involved in blood
coagulation
 Erythrocyte
 60-70% water
 ~35% solids (of which 95% is contributed by Hb and 5% by
stromal protein, lipids, phospholipids, cholesterol,
vitamins)
 RBCs contain large quantities of carbonic anhydrase; thus
transports large quantities of carbon dioxide from the
tissues to the lungs in the form of bicarbonate

 Hemoglobin is an excellent acid base buffer, therefore, the

RBCs are responsible for most of the acid base buffering
power of whole blood
 Mammalian RBCs are usually non-nucleated, biconcave,
circular discs with a central pale spot

 Shape differs in various species

 Dog, Cow, Sheep: Markedly biconcave
 Horse and cat: Shallow concaving
 Goat:Very shallow or flat surfaced erythrocytes
 Camel: Elliptical
 Deer: Sickle-shaped
 Amphibians and birds: Elliptical and nucleated
 Normal avian RBCs Normal canine RBCs

Normal feline RBCs

Normal deer RBCs Normal human RBCs

Normal camel RBCs Normal goat RBCs

 Average diameter of RBC: 4.1-7.5 µm

 Surface area varies from 57-67m2/kg body weight in mammals;

lowest in goat (smaller diameter) and highest in man (greater
diameter)

 Concentration of RBCs depends on various factors such as

interspecies, intraspecies, age, sex, environment, exercise,
nutritional status, climate and altitude
 Erythropoiesis
is your body's process of making red blood cells (erythrocytes).

 Hematopoiesis: The processes of formation of erythrocytes

(erythropoiesis), leukocytes (leukopoiesis) and platelets in the body

 From stem cell, the formation of reticulocyte takes about 72 hours and conversion
of reticulocyte to erythrocyte requires 48 hours; thus RBC formation requires 5 days

 Under appropriate stimulation, CFU-E progenitor cells produce proerythroblast

 Hb synthesis begins in polychromatophil erythroblast and maximum synthesis occurs

in orthochromatic erythroblast
 Hypoxia is the principle regulatory factor of erythropoietic activity of
the bone marrow
Hypoxia is low levels of oxygen in your body tissues. It causes symptoms like confusion, restlessness, difficulty
breathing, rapid heart rate, and bluish skin. Many chronic heart and lung conditions can put you at risk for hypoxia.

 Kidney produces 90% of erythropoietin and liver produces about 10%

hormone

 Erythropoietin stimulates hemopoietic stem cells of bone marrow to

produce the committed stem cells - proerythroblast
  Vitamin B12 and folic acid are essential for the maturation of erythrocytes
cobalamin

 Vitamin B12 is required for DNA synthesis

 Folic acid is required for RNA synthesis
 Macrocytic anemia is common in Vitamin B12 and folic acid deficiencies
Macrocytic anemia is a blood disorder that happens when your bone marrow produces abnormally large red blood cells.
B5 B3
 Thiamine (B1), Pantothenic acid, Nicotinic acid, Vitamin E and pyridoxine
(B6), riboflavin, biotin and ascorbic acid are essential for erythropoiesis
B2 B7 vitamin C
the type of anemia in which the circulating RBCs are smaller than the usual size of RBCs
(microcytic) and have decreased red color (hypochromic)
 Deficiency of Vitamin B6 causes microcytic hypochromic anaemia in pigs
 Normocytic anemia in swine and primates is due to Vitamin E deficiency
Normocytic anemia happens when you have fewer red blood cells than normal, and those blood cells don't have
the normal amount of hemoglobin.
 Minerals are essential for erythropoiesis

 Iron acts as an integral part of Hb which is essential for Hb

synthesis

 Copper acts as a co-factor in Hb synthesis

 Copper deficiency is common in pigs, which may interfere
with iron absorption and utilization

 In ruminants, cobalt plays a key role for the synthesis of

Vitamin B12 by the rumen bacteria
 A low percentage (1-3%) of erythrocytes in circulation has a network of
bluish threads within the cell and is called reticulocytes

 These cells are immature RBCs, which have entered into the
circulation at times of need

 In some diseases or due to excessive loss of blood or destruction of

RBCs, the reticulocyte number increases in circulation

 These cells have less or no O2 carrying capacity

 Energy is required for RBCs to:

 Maintain the shape and flexibility of the cell membrane

 Preserve high K+, low Na+ and low Ca2+ ions within the RBCs against the
concentration gradient of these ions of plasma

 Maintain iron in ferrous (Fe2+) state

 Generate reduced glutathione (anti-oxidant); this helps to maintain the Fe2+

state

 Generate 2,3-diphosphoglycerate (DPG) for O2 dissociation

 Mitochondria is absent in mature erythrocytes

 Thus, they derive their energy from glucose metabolism via

anaerobic Embden-Meyerhof (EM) pathway (90%) and oxidative
pentose cycle (10%) which produce NADH and NADPH

 Kreb's cycle is very much reduced in activity!

 Erythrocytes can change their shape when they pass through the
capillaries but they become less flexible when they reach the end of
their life span

 In most domestic animals, the bone marrow functions as a chief site of

erythrocyte destruction, whereas in man it is the spleen and in birds it
is the liver

 Two types of erythrocyte destruction takes place

1. Intravascular hemolysis
2. Extravascular hemolysis
 Intravascular hemolysis

 ~10% of aged RBCs undergo intravascular hemolysis within

the capillaries due to loss of compressibility of RBCs caused
by increased membrane permeability and osmotic change

 When this occurs the hemoglobin is released, which combines

with haptoglobulin, which is removed by the cells of the
mononuclear phagocytic system (MPS)
 Extravascular hemolysis

 ~90%of the aged RBCs are directly destroyed by the

mononuclear phagocytic system (MPS)

 The Hb and proteins are catabolized by the MPS cells; MPS

(also known as reticulo-endothelial system) includes the
histiocyte or macrophages, stellate or Kupffer cells of the sinu-
soids of the liver, spleen, mononuclear cells of bone marrow
and lymph nodes
 The globin of the Hb is degraded into amino acids and is
reutilized in the formation of new hemoglobin or other
proteins

 Iron removed from the heme is stored in the MPS cells in the
form of ferritin or hemosiderin and utilised for the synthesis of
hemoglobin or enters the plasma and combine with
apotransferrin to form transferrin which enters the bone
marrow to produce more erythrocytes
 Heme is converted into bile pigments, biliverdin (green pigment), and then reduced to
bilirubin (yellow pigment)

 Free bilirubin enters the plasma, binds with albumin and is transported to the liver, where
it is conjugated with glucuronic acid  secreted in bile

 Large intestinal bacteria reduce the bilirubin to urobilinogen, most of which are excreted
in faeces in the oxidised form of urobilin or stercobilin which impart colour to faeces

 Part of the urobilinogen is reabsorbed into the enterohepatic circulation and re-excreted in
bile

 Some of the urobilinogen in the plasma enters the kidneys to be excreted in urine as
urobilin
 Hemolysis caused by external agents:

 Blood parasites: Babesia, theileria, trypanosoma and

sarcocystis

 Chemicals: Copper, lead, nitrate and nitrite poisoning

Life span of erythrocytes

Species Life span (days)

Cattle 125-150

Sheep 140-150

Goat 125-150

Horse 140-150

Dog 100-120

Cat 70-80

Pig 51-79

Poultry 20-30
 Polycythemia

 Oligocythemia

 Anaemia
Polycythemia/erythrocytosis

 A condition of increased number of RBCs in the circulation; two types:

1. Physiological (secondary) polycythemia

 As a compensatory measure e.g. tissues become hypoxic because of

too little oxygen in the atmosphere, for example at high altitude or
because of failure of delivery of oxygen in the tissues as in cardiac
failure, then the blood forming organs automatically produce large
quantities of extra RBCs i.e. 30% above the normal
 Increased Hb requirement during heavy muscular exercise to
meet increased oxygen demand; in sport animals (racehorse,
hunting dogs) RBC elevation is a normal feature

 Increased environmental stress, the splenic contraction, and

increased RBC synthesis by the bone marrow cause increased
number of RBCs into the circulation

 Hemoconcentration due to water loss - vomiting, diarrhoea,

prolonged high fever and burns
2. Pathological polycythemia

 Due to decreased O2 supply to the tissue, chronic carbon

monoxide poisoning, myeloid (bone marrow) cancer,
pulmonary emphysema, repeated hemorrhage

 Polycythemia vera is the condition due to bone marrow cancer

(myeloid leukemia); it occurs as a result of genetic aberration
in the hemocytoblastic cell line that produces the blood cells
Oligocythemia

 Reduction in the number of erythrocytes in the circulation

1. Physiological oligocythemia occurs due to hemodilution e.g.

pregnancy

2. Pathological oligocythemia is also known as anaemia

Anaemia
 Abnormal reduction in the number of the erythrocytes or the
hemoglobin content in the blood or both

 Some causes:
 Excessive whole blood loss – hemorrhage
 Impaired RBC production and Hb synthesis - deficiency of Fe,
Cu, Vitamin B12 and folic acid
 Hemolytic - caused by blood parasites or drugs
(sulphanamides, antimalarial drugs)