CHAPTER 2 :BLOOD CIRCULATION and THE CARDIOVASCULAR SYSTEM

Blood serves as a transport medium.

Specific functions
Carries:
nutrients from the digestive tract to the tissues;
end products from cells to the excretory organs;
carbon dioxide from tissues to lungs;
secretions of endocrine glands throughout the body.
Regulate body temperature
Maintain constant concentration of water and electrolytes in the cells;
Regulate bodys hydrogen ion concentration;
Defend against microorganisms.
Homeostasis is the maintenance of uniformity and stability in this extracellular
fluid through physiological processes of diffusion, pressure gradients,
concentration gradients, active transport and regulatory mechanisms controlled by
the nervous and endocrine systems.
BLOOD CELLS, PLASMA and SERUM
Three classes of blood cells (corpuscles):
1. Erythrocytes (red blood cells)
2. Leukocytes (white cells)
3. Thrombocytes (platelets)
Hemoglobin is the pigment that contributes to the red color of the blood
contained in the erythrocytes.
Plasma is the yellow to colorless fluid in which all these cells are suspended. When
examined as a thin film, it is always colorless. Bilirubin is the pigment that
contributes to the color of plasma, although carotene and other pigments are
contributing factors.
In coagulation, blood lost from the body becomes a gelatinous mass, after which
the blood clot retracts, forcing from the clot a clear, watery fluid called serum
similar to plasma except that fibrinogen and other clotting factors have been
removed.
Plasma may be obtained by adding to whole blood an anticoagulant to
prevent clotting and letting the cells settle out, as they are heavier than plasma.
Centrifuging the blood hastens the settling of cells to obtain plasma more
readily.
ANTICOAGULANTS
FUNCTION: Anticoagulants are used to obtain blood samples free from clots for
transfusions and for analytical work.
Commonly used anticoagulants in transfusions:
1. Heparin, a conjugated polysaccharide, naturally produced by basophils (a
kind of leukocyte) in the blood and mast cells (part of the connective tissue
surrounding capillaries in the lungs and other organs) throughout the body.

Concentrations of this anticoagulant:

0.2 mg of heparin per ml of blood
1 mg of heparin of 100 500 ml of blood at 0C & 10 20 ml at room temp.
2. Sodium citrate,as the citrate combines with calcium ions of the plasma
forming an insoluble calcium salt.
Caution: One must be careful not to give too much citrate because citrate
can combine with sufficient calcium ions to produce tetany, and thus
interfere with the functioning of nerves and of skeletal and cardiac muscles.
Concentrations of this coagulant:
0.2 0.4 percent of blood
Similar salts are also used exceptpotassium salts because of the
possibility of producing a heart block.
Other useful coagulants in analytical work:
1. Sodium, potassium and ammonium salts of oxalates and fluorides.
2. Chelating compounds such as ethylenediaminetetraacetic acid (EDTA)
Note: Ammonium salts are not recommended when compounds containing
nitrogen are being determined quantitatively, because of nitrogen in the
anticoagulant.
Anticoagulants and their effects on cell size:
Analytical work:

Heparin or EDTA keep the size of erythrocytes constant, are important in

calculating mean corpuscular volume (MCV), mean corpuscular hemoglobin
(MCH) and mean hemoglobin corpuscular concentration (MCHC) as
diagnostic aids.
Ammonium salts increase the size
Potassium salts decrease the size
6 mg of ammonium oxalate&4 mg of potassium oxalate inhibited
coagulation of 5 ml blood & kept size constant (Heller & Paul (1934))

ERYTHROCYTES
Erythrocytes are biconcave, circular discs varying in diameter and thickness
according to species and nutritional status that are capable of undergoing changes
in shape while passing through capillary beds.
Appearance /structure in animals:

Mammals nonnucleated, nonmotile cells

Dog markedly concave
Cat & horse slightly concave
Ruminants (cattle, sheep, goats) and pigs discoid
Most animals below mammals elliptical and has nuclei.

Origin
Yolk sac produces nucleated red blood cells in early fetal development, later
involving liver, spleen and lymph nodes.
Bone marrow, place where the formation of red blood cells (erythropoiesis) takes
place after birth.
Under pathological conditions in postnatal life liver, spleen and lymph
nodes may again assume erythropoiesis.
Erythropoiesis (in bone marrow) goes on continuously and are poured into the
blood stream at a rate to balance the destruction of red cells.

TRANSPORT IS THOUGHT TO HAPPEN IN TWO WAYS

The entrance of the newly formed corpuscles into a bone marrow capillary
is more of penetration no stoma or opening is necessary. Erythrocyte being
nonmotile, enter the capillary by diapedesis( the passage of cell through the
unruptured blood vessel wall).
According to another view, erythrocytes develop intravascularly, as there are
two kinds of capillaries in the bone marrow:collapsed one which are
erthrogenic , and open ones through which the blood flows. The young ones are
forced into the blood stream as the erythrogenic capillaries become active and
patent.
Metarubricyte, is the nucleated bone marrow cell which non nucleated
erythrocyte is formed . The nucleus is lost by extrusion or absorption before the
corpuscles enter the blood stream.
Normally 1 3 percent of erythrocytes are reticulocytes(young red blood
cells that contains the remains of nucleus).
Pathological condition that increase their number in blood:

Hemorrhagic, hemolytic & other anemia when bone marrow is more

active in producing a great number of erythrocytes.
Exception: horse

Composition:
In adult

62 72 percent water
35 percent solids (most part hemoglobin; others: proteins, lipids, fat,
vitamins, glucose, enzymes and minerals)

Cations and anions (within & outside cell) help in establishing and maintaining
electrical gradients across cell membranes by the sodium pump, active transport
and diffusion (physiological processes that help in maintaining balanced
electrolytes in cells).

Sodium is the principal cation in extracellular fluid.Potassium is the principal

cation of erythrocytes of domestic animals except dog, cat, cattle, goats and
sheep.
Erythrocyte and potassium values:

Dog 8.7 and 107

Cat 5.9 and 103.7
Cow 21.8 and 79.8
Goat 18.4 and 93.2
Sheep 18.4 and 83.5, 64.2 and 15.6, 58.1 and 46.0 (depending on breed)
Pig 99.5 and 10.8
Chicken 97.3 and 7.1
Turkey 99.5 and 9.7

Size and hemoglobin content

There are many shortcomings in the measurement of the diameter of
erythrocytes, making it less important than cubic volume (which should be used
to measure cells.

The cells in the dry smear ( but in the moist state but losing water) are
smaller
Very few cells are measured from the blood sample taken
The depth of the cell or third dimension is left out of account

The following formulas will give mean corpuscular volume, hemoglobin, and
hemoglobin concentration in erythrocytes.

These formulas are an aid in diagnosing various types of anemias.

Iron deficiency anemia in mammals aremicrocytic type (very small cells)

MCVprovides the average size of cell size in cubic microns.

MCH expresses the average weight of hemoglobin present in the
erythrocytes.
MCHC gives the average percentage of the MCV which the hemoglobin
occupies.

Pernicious anemia in man is macrocytic, but this anemia does not occur in
animals.
Note: Error in counting erythrocytes contribute to variations in MCV and MCH.

Number
The number of red blood cells varies within species.
Plasma fluids are constantly being shifted across capillary walls, cell
counts vary between arterial and venous blood samples.
Factors that affect erythrocyte counts, hemoglobin concentration, PCV &
concentration of other blood constituents:

Age, sex, exercise, nutritional status, breed

lactation, pregnancy, egg production, stage of estrous cycle
excitement (release of epinephrine), blood volume (hemodilution or
hemoconcentration),
time of day, environmental temperature, altitude and other climatic factors.

Surface area
The surface area has great importance to the respiratory or gas-transport
function of the blood.
Total erythrocyte surface can be estimated when blood volume, erythrocyte
count, diameter, and thickness are known.
The estimated mean surface area per erythrocyte was calculated first from
the dimensions reported on dry films. This value was increased by 20 percent to
arrive at the estimated wet erythrocyte surface area.
The average thickness was 2 .
The figures for erythrocyte surface area compared to the body surface area
A man of average size has less than 2 sq m of body surface area, and a 500 kg
cow somewhat less than 5.

Tonicity
Rule: In order to keep erythrocytes constant in size they must remain in the
environment with the same osmolarity as blood plasma. When the osmotic

pressure of plasma is lowered sufficiently, hemolysis (laking) of erythrocytes

results.
3 Types of solutions:
1. hypotonic solutions osmotically weaker than plasma, causing hemolysis of
cells.
Plasma osmotic pressure may be lowered by adding hypotonic salt solutions
or water to blood. In such cases, water passes into the cells through osmosis
through its semipermeable membrane, causing the cell to swell. This results
in the stretching and eventual mechanical rupture of its membrane, with
hemoglobin passing through the surrounding medium.
2. Isotonic solutions similar to plasma in crystalloid osmotic pressure, into
which erythrocytes may be placed without resulting in cell volume changes.
3. Hypertonic solutions that exert a higher osmotic pressure than blood
plasma. They cause water to pass from the corpuscles by osmosis, and
shrinking of the corpuscles results. Such erythrocytes are said to crenated.
Physiological salt solution

also known as physiological saline

the isotonic solution of greatest interest.
an aqueous solution of sodium chloride usually containing 0.85-0.9 percent
sodium chloride.

Minimum resistance(osmotic resistance of the weakest corpuscles)

indicated by the concentration (%) of sodium chloride solution at which

hemolysis begins.

Maximum resistance(resistance of the strongest corpuscles)

indicated by the concentration at which complete hemolysis.

It Is not known whether hemolysis caused by lowering the osmotic pressure of the
plasma plays a part in the destruction of erythrocytes in the body under normal
conditions. However, knowledge of osmotic resistance is of practical importance in
preparing solutions for intravenous injection. Large amounts of water may be
injected into the blood stream at low rates without producing any significant
amount of hemolysis.
Processes that cause hemolysis of erythrocytes:

freezing and thawing

stirring and agitation
high temperatures
substances that lower surface tension (saponins, soaps and bile salts)
alcohol, ether, chloroform and acetone.

It is possible to calculate what percentage solution of a compound is isotonic with

erythrocytes by using the following formula:

Life Span
Length of life for erythrocytes:

man ranges from 90-140 days, averaging 120 days.

For several small laboratory animals- life span is much shorter

The half-life of erythrocytes is 17 days. The half-life of autologous (individuals

own erythrocytes) transfusions are longer than homologous (same specie)
transfusions.
Erythrocytes are destroyed in great numbers daily. The total number of
erythrocytes in the body of a 450 kg animal with a blood volume of 8 percent of
body weight is 300 trillion. If the average life of individual erythrocytes is
100 days, then 3 trillion must be destroyed (and formed) in the body
every day, or about 35 million every second.
Pathological conditions:

Iron-deficiency anemia erythrocytes become smaller in size

Microcytic type of anemia the result of younger cells not being released
into the circulating blood in sufficient quantities to replenish those being lost.

Fate of erythrocytes
Reticuloendothelial cells that destroy the old, exhausted erythrocytes:

Histiocytes
Macrophages
Clasmatocytes

Reticuloendothelial cells include the stellate or Kuppfer cells found in the liver,
spleen, bone marrow and lymph nodes.
As erythrocytes are destroyed, the iron-containing moiety of hemoglobin is
conserved and the pigmentary type is converted into bile pigment, an excretory
product.
Liver and spleen are important storehouses of the iron that is not immediately
used in the production of new hemoglobin.

The reticuloendothelial cells in different organs are important in the destruction of

erythrocytes. Red bone marrow is the principal place or erythrocyte destruction (
and bile pigment formation of dogs). Spleen for man; liver for rabbit, guinea pig
and birds.

Hematopoiesis
Hematopoiesis (the formation of red blood cells) is a continuous process.

Nutrients essential for this process:

Vitamin B12 (cyanocobalamin) functions in the maturation of erythrocytes

folic acid (pteroylglutamic acid)

Both vitamins act as coenzymes in the synthesis of nucleic acids or their

constituents, purine and pyrimidine bases.
Other vitamins that aid are:

pyridoxine, riboflavin, nicotinic acid

pantothenic acid, thiamine, biotin
ascorbic acid

When these vitamins are deficient, growth and development of erythrocytes are
impaired.
Pyridoxine deficiency in swine produces a microcytic, hypochromic anemia.
Other essentials:

minerals and amino acids

water and energy

The minerals most commonly needed:

Iron built into the hemoglobin molecule

Copper a coenzyme or catalyst in hemoglobin synthesis
Cobalt dietary essential for ruminants and is needed for bacterial synthesis
of vitamin b12 in the rumen.

Note:Cobalt given in excess will cause polycythemia. Though animals, if placed

in high altitudes may also show polycythemia because of the decreased oxygen
pressure (PO2).
Pathologic conditions that do not elicit the response for greater red cell
production.
Nongenerativeanemias like

Trichostrongylosis in cattle
Anemia associate with chronic infection
Neoplasia or other cachetic diseases

Pathologic conditions that elicit the responsefor greater red cell production.

Hemorrhage
Hemolytic diseases
Blood-sucking parasites
Nutritional deficiencies

In hypoxic conditions (tissues are not supplied with sufficient oxygen) caused by
inadequate number or improper functioning of erythrocytes) resulting in the
release of erythropoietin, hemopoetin or erythrocyte maturing factor to
blood plasma that stimulates hematopoiesis.Thekidney has been cited as
producing this, but hypoxic cells in general may produce it.

PACKED CELL VOLUME

The volume of cells in the circulating blood is usually less than the plasma volume.
Hematocrit test utilizing a centrifuge in which is placed a small tube containing
a sample of blood with the appropriate kind and quantity of anticoagulant.
In the past
The Wintrobe hematocrit tube is centrifuged at 3000 rpm for 30 minutes; then the
percentage of packed red cells is read from a scale or calculated.
As a rule in normal animals the volume of packed red cells (PCV) is directly related
to the erythrocyte count and hemoglobin content.
With this rule, it became a common practice in some laboratories to allow the
hematocrit tube containing blood to stand vertically for an hour before
centrifugation in order to obating the erythrocyte sedimentation rate.
In Recent years
The microhematocrit centrifuge and tubes (with or without heparin) have been
used widely to save time, values being obtained in 5 minutes.
The PCV is expressed as a percent volume of packed cells in whole blood after
centrifugation.
Hemoconcentration due to dehydration, asphyxia, or excitement causing
release of erythrocytes in the spleen can result in abnormally high PCV values. In
excitement, the epinephrine causes splenic contractions.

HEMOGLOBIN
Hemoglobin, the pigment in erythrocytes, is a complex, iron-containing,
conjugated protein composed of a pigment and a simple protein.

Globin the protein, a histone.

Heme causes the red color, a metallic compound containing an iron atom.

Biosynthesis of hemoglobin starts in the erythroblasts and continues in the

subsequent stages of development.
Four heme molecules unit with oneglobin to form hemoglobin.
The molecular weights of hemoglobin from most species are reported to vary from
66,000 to 69,000. Based on the iron content of hemoglobin as 0.334 percent and
the atomic weight of iron being 55.84, a value of 16,700 as the minimal molecular
weight of hemoglobin is obtained.
Hemoglobinuria occurs when erythrocytes are hemolyzed in the blood stream by
protozoa, toxins or chemical agents, releasing hemoglobin into the plasma that is
permitted passage by the basement membrane of the kidneys.

Fetal and Adult hemoglobin

At a given oxygen tension, fetal hemoglobin will take up more oxygen
than adult hemoglobin.
The passage of oxygen from the mother to the fetus is by diffusion, the tension of
oxygen in the blood of the umbilical vein is same as the venous blood on the
maternal side of the placenta.
The production of fetal hemoglobin depends upon presence of amino acids at
site of formation or origin of erythrocytes.
Infants may be born with one half to three fourths of their hemoglobin of the
fetal type. By the end of the first year, fetal type of hemoglobin may reach 1
percent. The animal retains the ability to make fetal hemoglobin in adult life.

Amount
The amount of hemoglobin in the blood is expressed as g/100 ml of blood.
As a rule in most mammals normal blood hemoglobin values are between 13 and
15 g/100 ml. Exception: lactating cows. Cold-blooded: usually lower, being 12-13.
Excitement elevates erythrocyte numbers per unit of volume, easily
demonstrated with anesthesized dog after injection of epinephrine.
The hemoglobin concentration in the avian blood is more difficult to
determine. A method has been devised to determine hemoglobin in bloof of
chickens and values obtained are similar to those when iron method is used.
Normal hemoglobin concentration in chicken ranges from 6.5 to 9 g/100 ml by this
method.

Conditions that lower the oxygen content of blood, like elevated barometric
pressure, cause an increase in the production of hemoglobin and the number of
erythrocytes and vice versa.