Symposium on Clinical Hematology

The Leukocytes

Joseph G. Zinkl, D.V.M., Ph.D.*

The blood leukocytes or white blood cells are the neutrophil, eosin-
ophil, basophil, lymphocyte, and monocyte. The first three of these cells
are collectively called granulocytes because they contain many distinct
granules within their cytoplasm; they also have segmented nuclei. Leu-
kocytes other than these are observed in blood in some conditions. Im-
mature forms of neutrophils, including neutrophilic bands, myelocytes,
and metamyelocytes, may be found in the blood of animals with inflam-
matory diseases. Occasionally plasma cells or plasmacytoid lymphocytes
may be found in the blood of animals mounting an antibody response.
Leukemic processes are often characterized by immature and bizarre
forms of the major blood cell types, as well as the presence of other
cells, such as mast cells, in the blood.

FUNCTIONS OF LEUKOCYTES

Neutrophils
The primary function of neutrophils is to kill and digest microor-
ganisms that invade the body. They perform this function mainly in the
tissues; therefore, they have abilities that allow them to leave the blood
and get into the tissues. After leaving the blood the neutrophil moves
toward the microorganism, recognizes it as a detrimental substance, in-
gests or phagocytizes it, and kills and digests it (Fig. 1).
Neutrophils newly released from the bone marrow have a blood
half-life (T 112 ) of six to ten hours. Generally they leave the blood in a
random manner; however it has recently been suggested that time spent
in the blood stream helps them acquire additional abilities useful in the
tissues. Perhaps, at least in healthy animals, the older cells marginate
and leave the blood preferentially to the younger cells. The blood neu-

*Assistant Professor, Department of Clinical Pathology, University of California, Davis

School of Veterinary Medicine, Davis, California; Diplomate, American College of
Veterinary Pathologists

l"eterinary Clinics of North America: Small Animal Practice- Vol. 11, No.2, May 1981 237
 238 JosEPH G. ZrNKL

2.
f)

* I.

BLOOD

TISSUE
Figure l. Neutrophils function by (1) migrating from blood to tissues, (2 and 3)
phagocytizing opsinized organisms, and (4) forming a phagosome. Lysosomes discharge
their contents into the phagolysosome (5), which kills the organism (6). (7) The killed
organism may be digested or discharged from the cell.

trophils are divided into two groups or pools: marginated and circulat-
ing. It is thought that there is relatively free exchange of cells between
the two pools, but recent evidence suggests that marginated cells con-
tinue to mature and may have increased abilities for combating foreign
substances compared to those of the circulating poolY
After adhering to the endothelial cells, neutrophils migrate between
cell spaces into the tissues. They do this in response to positive chem-
otactic (attractant) substances, of which there are many. Some suchas
cell wall lipids and formyl-methionyl peptides are bacterial
products. 19• 36 • 39 Complement factors C5 a and C3 a, derived from com-
plement activation, are chemotactic to neutrophils. 19 Products of the
kinin and coagulation system also attract neutrophils. Some prostaglan-
dins and cyclic 3' ,5'-AMP are chemoattractants for neutrophils. 19 Re-
lease substances from cells that are dying or undergoing degeneration
attract neutrophils to areas of necrosis. Fragments of IgM and collagen
also attract neutrophils. 19 Thus, it is evident that a variety of reactions
can cause the production or release of substances that attract neutrophils
to microorganisms and damaged tissue.
Once the neutrophil arrives at the site of the microorganism, it is
necessary for it to recognize the substance that must be disposed. Op-
sonization is the process by which objects are labelled as foreign. This
process involves the coating of the object with complement factors and
LEUKOCYTES 239
immunoglobulin. Complement factor Cab and IgG are the most impor-
tant opsonins. The surface of the neutrophil has receptors that attach
to the opsonins and bind the neutrophil with the foreign particle. In
this manner the neutrophil both recognizes the foreign substances and
begins the process of phagocytosis. 19• 24
Microfibrilar action within the neutrophil begins the process of in-
ternalization of the particle. The cell membrane invaginates and sur-
rounds the particle. Eventually the membrane pinches off from the cell
surface and a membrane-bound particle or phagosome is found free in
the cytoplasm. At this stage the granules of the neutrophil fuse with the
phagosome to form a phagolysosome. The process allows the antimi-
crobial substances of the neutrophil to make contact with the foreign
particle. The antimicrobial substances kill and partially digest the par-
ticle. 19, 24
Phagocytosis by neutrophils is accompanied by antimicrobial activity
which, in large part, is due to metabolic changes. Increased consumption
of oxygen and utilization of glucose occur. Hexose monophosphate ac-
tivity is markedly increased. This "metabolic burst" causes the produc-
tion of oxygenated substances with antimicrobial activity. Hydrogen per-
oxidase (H 2 0 2 ), superoxide anion (O-), hydroxyl radical (OH-), and
singlet oxygen (102 ) are products of the "metabolic burst." These prod-
ucts act as antimicrobial substance in consort with decreased pH, the
enzyme myeloproxidase, and halide ions (Cl- and J-). 19
These reactions occur within the phagolysosome since they are toxic
to neutrophils as well as to bacteria. However, some of the oxygenated
products can diffuse across the phagolysosomal membrane and enter
the cytosol. The neutrophil contains systems within its cytoplasm to neu-
tralize these products. The hexose monophosphate pathway maintains
NADPH production necessary to maintain glutathione in the reduced
state. Reduced glut:tthione detoxifies hydrogen peroxide. Superoxide
dismutase converts superoxide radical to hydrogen peroxide, which is
then detoxified by glutathione peroxidase. 19
Antimicrobial mechanisms other than those dependent on the me-
tabolism of oxygen are also present in neutrophils. The physical isola-
tion of bacteria away from serum and its rich supply of nutrients may
be bacteriostatic. Lysosomal degranulation delivers into the phagosome
a variety of antibacterial substances including lysozyme, neutral and acid
hydrolases, cationic proteins, lactoferrin, and collagenase. 19 Many bac-
teria are not only killed but also digested by these neutrophil enzymes
and products.
The killed bacteria or the digested products are released from the
cell by exocytosis when the vacuole fuses with the cell membrane and
evaginates. The neutrophil may also die and rupture, or it may be phag-
ocytized and degraded by a macrophage. Neutrophils that have mi-
grated into tissues (perhaps in a surveillance role) but have not partic-
ipated in an antimicrobial reaction, die after several days. If they have
240 JosEPH G. ZINKL

migrated into a tissue with an epithelial surface, they may be lost

through the mucous membranes. Many circulating neutrophils are
probably disposed of in the spleen. 18
Eosinophils
Eosinophils have some abilities similar to those of neutrophils. They
respond to chemotactic agents, are phagocytic, and produce the same
oxygen metabolites. The eosinophil is primarily a cell of tissues, having
a blood-to-tissue ratio of about 1:300. 4 High concentrations of eosino-
phils are present in subepithelial areas.
Eosinophils stain with acid dyes because of a substance in their
granules called major basic protein. 4 The granules contain many other
substances, many of which are similar to those of neutrophils.
Chemotaxis by eosinophils is similar to that by neutrophils in that
many substances that attract neutrophils also attract eosinophils. Eosin-
ophils also are attracted by a substance called eosinophilic chemotactic
factor, which is produced by mast cells and basophils. 4
Eosinophils are capable of phagocytosis, although apparently they
are less efficient than neutrophils. Some substances such as mast cell
granules and immune complexes are easily phagocytized. Eosinophils
discharge their granules easily, suggesting that their contents are im-
portant extracellularly. Eosinophils may be especially active in killing
parasites. Perhaps the ability to undergo exocytosis in proximity to the
parasite releases killing substances directly upon the parasite. 4 This abil-
ity would overcome the obvious difficulty of ingesting such large invad-
ers.
The major function of eosinophils probably is to modulate and
confine the inflammatory response. 4 They do this by neutralizing in-
flammatory substances such as histamine and by phagocytizing immune
complexes. In inflammation the highest concentration of eosinophils are
in the periphery of the lesion. This "bystander" location is ideal for
containing the reaction. Presumably the eosinophil neutralizes media-
tors diffusing from the center of the lesion. Eosinophils inactivate sub-
stances of mast cell granules such as histamine. Eosinophils may also
produce certain prostaglandins that inhibit mast cell degeneration. The
kinin system may also be inhibited by eosinophil products.
Basophils
Basophils are distinctive because of their granules, which stain with
metachromatic dyes. In blood they are in low numbers, but they are
found in many tissues. Morphologically and functionally they are similar
to mast cells, however these two cell types probably arise from different
precursors. Basophils and mast cells have surface receptors for IgE.
Degranulation of basophils occurs in both antibody and cell-mediated
hypersensitive reactions. In the process they release potent pharmaco-
logic mediators such as histamine, heparin, and other substances that
act as initiators of inflammation. 10
LEUKOCYTES 241
Basophils are able to migrate. A factor produced by lymphocytes
is especially chemotactic for basophils. This substance may be elaborated
in high concentrations by primed lymphocytes of hypersensitive ani-
mals.10
Compared to neutrophils and eosinophils, basophils have limited,
poorly developed phagocytic ability. Basophils are able to undergo ex-
ocytosis and probably are continuously releasing small amounts of sub-
stances.
In co~unction with mast cells, basophils elicit and maintain the
inflammatory reaction. They play a role in the pathogenesis of imme-
diate hypersensitivity and allergic reactions. Antigen-derived release of
basophilic substances is dependent upon IgE antibody coating of the
cells.
Monocytes
The monocyte is an immature cell that matures after leaving the
blood to become the tissue macrophage. The macrophage acts in both
the phagocytic and the immunologic systems. As monocytes mature they
increase in size, dense granules (lysosomes) and other organelles become
more abundant, and glycolytic ability increases markedly.
The monocyte-macrophage has three well defined functions: (1)
defense against microorganisms, (2) removal of damaged cells and de-
bris, and (3) interactions with the immune system. They also play a role
in other less well defined activities, such as in reactions against tumors,
in the control of granulopoiesis and erythropoiesis, and in wound heal-
ing and bone remodeling. 6
Macrophages phagocytize substances similarly to neutrophils. They
phagocytize large particles easier than neutrophils do. Macrophages are
probably more efficitmt than neutrophils when the particle load is high. 6
Macrophages form the principal defense system against certain
classes or organisms. Bacteria, which exist primarily intracellularly, and
many fungi are killed or their growth is limited by macrophages. The
mechanisms for killing organism probably are similar to those of neu-
trophils.
Macrophages are also involved in the afferent and the efferent
limbs of the immune system. In the afferent or inductive phase of the
immune response, macrophages process a small proportion of the phag-
ocytized substance in a manner that enhances the immunogenicity of
the substances. The macrophage presents this processed material to B
and T lymphocytes. Ultimately, B and T effector and memory cells are
produced.
Lymphocytes appear to reciprocate with macrophages in the effer-
ent limb of immunity. Stimulated T lymphocytes produce lymphokines
that have, among other actions, the ability to activate macrophages. Ac-
tivated macrophages have increased abilities to kill organisms. This abil-
ity is nonspecific, thus activated macrophage activity is directed toward
a variety of agents in addition to the agent that originally stimulated the
242 jOSEPH G. ZINKL

T lymphocyte. 6 This action is the basis for using highly immunogenic

substances such as BCG in antitumor therapy.
Antitumor activity and control of hematopoiesis are other activities·
that have been suggested for macrophages. Macrophages seem to have
an ability to kill certain tumor cells. Control of granulopoiesis may be
due to macrophages producing a substance called colony-stimulating
activity (CSA). In erythropoiesis, macrophages are important in trans-
porting iron to developing cells as well as removing excess iron from
more mature erythropoietic cells. Sequestration of iron in macrophages
during chronic disease appears to be the mechanism of the anemia of
chronic disease.
Lymphocytes
Two distinct types of lymphocytes are recognized, the B cells and
the T cells, based upon the site of their early processing and certain in
vitro reactions. These cells are responsible for the dual immune system,
humoral immunity (B cells) and cell-mediated immunity (T cells). Hu-
moral immunity is due to antibody production and acts primarily against
the extracellular phases of bacterial and viral infections. Cellular im-
munity acts against fungi, intracellular pathogens, and foreign tissue
such as transplants and cancer cells.
When primitive lymphocytes are processed by the bursa of Fabricius
in birds or its equivalent in mammals (probably the bone marrow), it
becomes a B lymphocyte. Each B lymphocyte contains on its surface
immunoglobulins that act as receptors for specific antigens. When an
antigen binds to a B cell receptor, a humoral response follows. A few
to many lymphocytes may contain similar receptors, which are known
as clones. There are many clones of B lymphocytes, which explains the
ability of the immune system to respond to many antigens. 6 • 25
When an animal is exposed to antigens, the B lymphocytes begin
to proliferate and differentiate in a process called clonal expansion.
Some of the progeny become effector cells that produce antibody. These
are recognized in tissues as plasma cells. Another group of stimulated
B cells becomes memory cells. These cells may exist for years, as opposed
to a few days for effector cells, and are responsible for the anamnestic
response that occurs when reexposure to the same antigen occurs.
T lymphocytes are produced by processing primitive cells in the
thymus. The process of stimulation and differentiation is similar to that
of B cells. The nature of the membrane receptors is unknown. Effector
cells of the T lymphocyte series are divided into several classes. Among
these are helper cells and suppressor cells, which regulate the humoral
response. "Killer" cells or cytotoxic T cells, which directly eliminate for-
eign cells, are also produced. Several other kinds ofT cells with different
activities also exist. 25
LEuKOCYTES 243
KINETICS OF LEUKOCYTES

The leukocytes of the blood represent only a part of the total body
leukocytes. The white cells are distributed throughout the body. Reac-
tions in various parts of the body can have profound effects on the
easily sampled blood leukocyte count. An understanding of the sites of
production, transportation, and utilization of leukocytes can improve
the ability of the veterinarian to understand the effects of diseases on
blood leukocytes.
Granulocytes
The granulocytes are found in three main areas: the bone marrow,
blood, and tissues (Fig. 2). The bone marrow contains a precursor pool
and a reserve pool of cells. The precursor pool is made up of those cells
able to undergo mitosis: myeloblasts, promyelocytes, and myelocytes.
The reserve pool is divided into two smaller pools: the maturation and
the storage pools. The maturation pool consists of metamyelocytes and
band cells. The storage pool also contains band neutrophils as well as
the mature neutrophils. All the cells of the reserve pool can be mobilized
rapidly when needed. 10
In the blood there are two pools of granulocytes: the marginal and
the circulating pools. The marginal pool consists of the granulocytes
adhered to the endothelium. Many of these cells are in the first stages
of migration into tissues, others may be in capillaries with little or no
blood flow, and others may be sequestered in the spleen. Many of these
cells enter the circulating pool spontaneously, or they may enter under
certain physiologic or pathologic stimuli. Thus, in the blood granulocyte
pool, there is relatively free exchange between the marginal and circu-
lating compartments. It has recently been suggested that marginated
cells are more mature' than circulating cells and that time spent in the
blood is necessary for these cells to acquire some refinements of their
abilities. 11
The end stage of the granulocyte is in the tissues. There is no
recirculation from the tissues to the blood.
The maturation of granulocytes takes four to seven days. The time
spent in the blood is six to ten hours. In the dog, about half of the

BONE MARROW

--
r-- RESERVE POOL BLOOD

•
MARGINAL
POOL
PROLIFERATION
POOL
MATURATION
POOL r+ STORAGE
POOL CIRCULATING
TISSUE
POOL
Figure 2. Diagrammatic representation of the production and utilization of neutro-
phils.
244 JosEPH G. ZINKL

neutrophils are in the blood pool; in the cat there are about three times
more neutrophils in the marginal pool than in the circulating pool.
Neutrophils may exist in the tissues for up to five days; however, this
is usually shortened markedly when toxic products are present.
Eosinophil kinetics are similar to those of neutrophils. The blood-
to-tissue ratio of eosinophils is about 1 :300. The fate of eosinophils in
tissues is not known. It is likely that many are shed from mucosal sur-
face, since they tend to localize in subepithelial areas. 4
Little is known about the kinetics of basophils. Basophils seem to
behave similarly to eosinophils.
Monocytes
The kinetics of monocytes is similar to that of neutrophils in that
monocytes leave the site of production, the bone marrow, enter the
blood, and egress to the tissues. They do not recirculate. A reserve bone
marrow pool does not appear to exist for monocytes, probably because
the circulating monocyte is an immature cell that matures in the tissues
and does not need a site such as the marrow to mature. In the blood
monocyte pool, about one-quarter of the cells are in the circulating
compartment and the remaining cells are in the marginal pool. The
blood half-life of monocytes is about ten hours. The ratio of blood
monocytes to tissue monocytes is about 1 :50. 6
Lymphocytes
Unlike granulocytes and monocytes, lymphocytes circulate through
blood and tissue repeatedly. Tissue lymphocytes return to the blood by
the lymphatics, entering one of the large systemic veins near the thoracic
inlet by way of the thoracic duct or the right lymphatic duct.
There are several peripheral lymphatic tissues in which many lym-
phocytes are produced and reside. Among these tissues are the lymph
nodes, spleen, subepithelial lymphoid tissue, and bone marrow. These
are often the site of lymphocyte-antigen interaction. Within the periph-
eral tissues T and B cells are located in specific sites.
In lymph nodes the T and B cells leave the blood at postcapillary
venules located in the cortex. These venules have a high endothelium
that is specialized to aid the passing of cells into tissues. B cells migrate
to follicles in the cortex; T cells occupy the diffuse cortex. Both cells
begin recirculating by passing through the medullary sinus to enter
efferent lymphatics (Fig. 3).
In the lymph nodes, macrophages filter antigenic material and pre-
sent it to B and T cells circulating through the node. An immune re-
sponse results in expansion of B and T cell clones and the production
of effector and memory cells. When this occurs primary follicles become
secondary follicles. The activated follicles are composed of a densely
cellular mantle, similar to the primary follicle, containing mostly mature
lymphocytes, and a germinal center that contains lymphocytes, lym-
LEUKOCYTES 245
Poslcopil/ory venules

Subcortical space

PrJmory
follicle

Corticomedullory
Junction Efferent lymphatic
Figure 3. Diagrammatic cross-section of a lymph node.

phoblasts, and debris-laden macrophages. Medullary cords running to-

ward the hilus contain large lymphocytes and plasma cells. Medullary
sinuses separating the medullary cords converge to form efferent lym-
phatics at the hilus. Circulating lymphocytes enter the tissues from the
medullary cords (B cells) or the diffuse cortex (T cells), after passing
through the cortex. 25
In the spleen the T and B cells also occupy specific domains (Fig.
4). Branches of trabecular arteries, called central arteries, enter the
splenic parenchyma. These arteries are surrounded by lymphoid
sheaths containing T lymphocytes. In addition, eccentrically placed pri-

SPLEEN

Red Pulp Cords

Figure 4. Diagrammatic cross-section of the spleen.

246 JosEPH G. ZINKL

mary and secondary follicles are found along the periarterial sheath.
These are the location of B cells. Adjacent to this "white pulp" is the
marginal zone containing macrophages. The central artery sends many
branches into the periarterial zone to the marginal zone. The central
artery terminates by branching into long, slender arteries (penicellar
arteries). The capillaries of these arteries are surrounded by an ellipsoid
sheath of macrophages. These capillaries open up into the reticular
network or red pulp of the spleen. Macrophages of the marginal zone
act with BandT cells in this area to produce an immunologic response. 25
Among the functions of the macro phages of the red pulp is the removal
of effete red cells and red cell inclusions.
Lymphocytes are present in subepithelial locations of the gastroin-
testinal, respiratory, and genitourinary systems. The structure of these
tissues varies from single nodules to more organized structures such as
the tonsils. These aggegates also contain B and T lymphocytes. The B
cells are in follicles, whereas the T cells are in more diffuse lymphoid
areas. Many B lymphocytes seem to "home" specifically to subepithelial
locations where they transform into IgA-producing plasma cells. Two
IgA molecules combine with an epithelial product called secretory com-
ponent, which facilitates transmucosal transport. Secretory IgA prevents
bacterial attachment to mucosal surfaces, binds enteric toxins, and pro-
tects the mucosal surface.
The bone marrow contains T and B lymphocytes in the postnatal
animal. The marrow functions as a peripheral lymphoid organ in ad-
dition to being the source of lymphoid stem cells and probably acting
as the equivalent of the bursa in mammals. Increased numbers of lym-
phocytes are seen in some conditions. In normal cats 10 to 15 per cent
of the marrow cells may be lympho1=ytes.
The blood lymphocyte count is influenced by conditions within the
peripheral organs. Changes in these organs or in the circulation of the
lymphocytes can alter the blood lymphocyte count. In healthy animals
it is estimated that about 70 per cent of the blood lymphocytes are T
cells and 30 per cent are B cells. Recent studies suggest that some of
the blood lymphocytes are not classifiable, however. 2

LEUKOPENIA AND LEUKOCYTOSIS

Quantitative and qualitative changes in total leukocyte numbers or

in specific cells occur in many conditions. These changes may be accom-
panied by morphologic and functional abnormalities.
Neutropenia
Leukopenia in dogs and cats is caused primarily by neutropenia
(Fig. 5). Neutropenia is a condition of the circulating pool and may or
may not indicate a similar condition in other pools. Neutropenia indi-
LEUKOCYTES 247

Tissue
Normal

Decreased
Proliferation

Ineffective
Proliferation

Early
- ---§Bj- Increased
Periphero I
Utilization

Late
Increased
Peripheral
Utilization

Marginal
Pool
Shift

Decreased
Storage
Egress

Figure 5. The mechanisms of neutropenia. P.P., proliferative pool; R.P., reserve

pool; Ma.P., maturation pool; S.P., storage pool; M.P., marginal pool; C.P., circulating
pool.

cates that there is either decreased input to the circulating pool or in-
creased egress from the pool. 15
Neutropenia -due to decreased input from the marrow may be
caused by decreased production of cells in the proliferative pool and
decreased egress from the storage pool. Proliferative pool defects are
caused by decreased numbers of granulocyte precursors or ineffective
proliferation.
Among the causes of decreased granulocyte precursors are malig-
nant cell invasion of the marrow and myelofibrosis. Drug-induced neu-
tropenia is usually of this type. Cancer chemotherapeutic agents pro-
duce neutropenia by this mechanism. Hematologic recovery may occur
upon cessation of drug intake; however, it may be extremely prolonged
or irreversible damage may occur. Bone marrow smears show reduced
numbers of myeloid elements.
Ineffective proliferation is a rare cause of neutropenia. Some
chemotherapeutic agents, such as methotrexate, that block folic acid
utilization cause neutropenia in this way. Smears of bone marrow show
increased immature granulocyte precursors.
Decreased storage pool egress is a rare cause of neutropenia. Neu-
248 JosEPH G. ZINKL

tropenia due to decreased pool egress is usually transient and probably

reflects a lag in the shifting from reserve pool cells to the blood pool.
Neutropenia due to shifting of cells from the circulating pool to the
marginal pool is not uncommon. The neutropenia of endotoxic shock
is, at least in part, due to increased numbers of neutrophils adhering to
the endothelium. The sluggish blood flow of shock may cause neutro-
penia by producing conditions that allow cells longer periods of time to
become adhered to endothelium.
In dogs and cats the most common cause of neutropenia is in-
creased utilization. Usually this occurs early in infectious processes in
which cells sequester near the agent. If the condition persists, continued
utilization of cells depletes the reserve pool and recovery may require
several days. Continued neutropenia in infections suggests that a viru-
lent organism is the cause, and the prognosis is poor. Viral diseases such
as canine distemper may in part produce neutropenia by destroying
neutrophils. When there is destruction or utilization of blood neutro-
phils, the bone marrow attempts to compensate and a left shift will be
found.
Neutrophilia
Several mechanisms cause neutrophilia in dogs and cats (Fig. 6).
Shift neutrophilia occurs when cells in the marginal pool shift to the
peripheral pool. 14 Storage pool shifts occurring with stress and after
steroid administration cause neutrophilia. Another consequence of ste-
roids is to decrease egress of cells from the blood.

Tissues Normal

Circulatory
Pool
Shift

D-DDEl Decreased
Egress

Increased
Bone
Marrow
Proliferation

Figure 6. The mechanisms of neutrophilia. P.P., proliferative pool; R.P., reserve

pool; Ma.P., maturation pool; S.P., storage pool; M.P., marginal pool; C.P., circulating
pool.
LEUKOCYTES 249
Inflammation causes neutrophilia due to increased proliferation
and persistent shifting of neutrophils from the storage pool to the
blood. 14 When extremely elevated neutrophil counts are found, the most
likely cause is inflammation, usually of infectious etiology. This is often
accompanied by a left shift. Moderately elevated counts can be due
either to physiologic mechanisms or to modest tissue damage.
A high neutrophil count with a left shift suggests that the bone
marrow has the ability to meet the needs of the animal in combating an
inflammatory agent. In contrast, left shift with low or normal total neu-
trophil count suggests an inability to meet the needs of the animal.
Eosinophils
Eosinopenia is rarely recognized because the low normal values,
based upon 200 cell differential counts, are zero. Eosinophil counts con-
ducted by eosinophil-counting techniques suggest that there are at least
two different mechanisms of eosinopenia: stress and inflammation. 3 The
eosinopenia of stress is well recognized and is mediated through en-
dogenous release of corticosteroids. Eosinopenia of inflammation is me-
diated by some other mechanism since adrenalectomy does not abolish
the eosinopenia of inflammation. 3 Perhaps this effect is due to eosino-
phils responding chemotactically to products of inflammation.
Eosinophilia may be encountered in several conditions such as skin
diseases, heartworm infestation, and pulmonary diseases. Parasitic in-
festations may increase the eosinophil count considerably, but it is
thought that this response occurs only during the parasite's migratory
phases. 4 Pulmonary infiltration by eosinophils can occur without clinical
signs referable to the respiratory system. Eosinophilia may be present,
however. Severe pulmonary disease often has elevated eosinophil
counts, and some of the most marked eosinophilias occur with respi-
ratory disease. Perhaps many of these diseases have an allergic etiology.
Basophils
Basophilia occasionally occurs in dogs and cats, but the conditions
producing it are difficult to relate to each other and basophilia does not
seem to be a consistent finding in any disease of dogs and cats. In hu-
mans basophilia is often associated with the precancerous or early stages
of granulocytic leukemia. Basopenia is not recognized in routine leu-
kograms.
Monocytes
Monocytosis may be seen in several conditions and does not nec-
essarily indicate that a disease is chronic. Monocytosis occurs in stress
and after steroid administration in dogs but in cats. 35 Monocytosis may
be found in diseases in which necrosis is present. Autoimmune diseases
have markedly increased monocyte counts. Autoimmune hemolytic
anemia is usually associated with markedly elevated monocyte counts.
250 JosEPH G. ZINKL

Trauma in dogs usually causes the monocyte count to be elevated. Frac-

tures, ruptured intervertebral discs, and other lesions involving bones
cause the monocyte count to be increased.
Dogs with diseases of fungal etiology may have elevated monocyte
counts. These diseases are characterized by granulomatous inflamma-
tion. People with neutrophil dysfunction, such as chronic granuloma-
tous disease, have elevated monocyte counts in an attempt to compen-
sate for the lack of functional neutrophils. Monocytosis may also occur
in Irish Setters with granulocytopathy, 27 • 28 but detailed hematologic
studies of this condition have not been published.
As with eosinopenia and basopenia, monocytopenia is not easily
recognized in routine leukograms. Little is known concerning its causes
or significance.
Lymphocytes
Lymphocytosis occurs occasionally in dogs and cats. It is seen in
physiologic leukocytosis. Other nonneoplastic causes of lymphocytosis
include the later stages of resolving infectious diseases (especially those
of viral etiology), autoimmune disease, and rarely, after vaccination.
Lymphopenia is an extremely common finding in canine and feline
leukograms. The most common cause of lymphopenia is stress, perhaps
due to lysis and decreased production of lymphocytes. Persistent lym-
phopenia indicates that the animal is undergoing continued stress and
suggests a guarded or poor prognosis.
Disruption of lymphocyte circulation can cause lymphopenia. Dogs
and cats with rupture of the thoracic duct may have low blood lympho-
cyte counts. Lymphangiectasia may cause loss of lymphocytes and
plasma proteins into the intestine.
People and animals, including ,Arabian foals, with immunodefi-
ciency diseases often have lymphopenia. Similar conditions have not
been described in dogs and cats. Several reports of Pneumocystis pneu-
monia in Miniature Dachshunds suggest that some members of this
breed may have an immunodeficiency disease because it is widely re-
garded that an animal's immune system must be compromised before
Pneumocystis carinii causes disease. 7 • 17

MORPHOLOGIC ABNORMALITIES OF LEUKOCYTES

Neutrophils
The most common morphologic abnormality observed in blood
smears is the toxic neutrophil. This term is applied to a variety of mor-
phologic abnormalities. There are two basic causes of the changes seen
in toxic cells: asynchronous maturation of the nucleus and cytoplasm,
and degenerative effects in the blood (Table 1). Cytoplasmic basophilia.
LEUKOCYTES 251
Table 1. Characteristics of Toxic Neutrophils
CHARACTERISTIC MORPHOLOGY AND CAUSE

Basophilia Persistence of the bluish cytoplasm usually

Toxic Granulation
Foamy or Vacuolated Cytoplasm
Dohle Bodies
Giant Neutrophils, Bands, and
Metamyelocytes
found in immature neutrophilic cells such as
metamyelocyte and myelocytes
Primary granules retain some of their ability to
stain azurophilic or reddish in mature cells;
blue-black granules may also occur
Toxic products cause lysosomal release and
development of vacuoles
Small (up to I p.m) areas of basophilia
representing aggregates of endoplasmic
reticulum
Very large cells, perhaps polyploidal, that may
represent skipped cell division; particularly
present in cats

Dohle bodies, toxic granulation, and giant, bizzare cells are probably
produced as a maturation defect. Foamy and vacuolated cytoplasm
probably are due to toxic products causing degenerative effects in cir-
culating cells. Toxic cells have diminished functional abilities, 1• 19 and
significant numbers of these cells may compromise an animal's resis-
tance. Animals with toxic regenerative left shifts probably can compen-
sate more than adequately for the decreased ability by the increased cell
counts, but animals with degenerative left shift might be compromised
by both the lack of adequate cell numbers and the poor ability of the
cells.
Another morphologic abnormality of neutrophils is Pelger-Hiiet
anomaly (Fig. 7). 19 In this condition the nucleus fails to undergo ade-
quate segmentation although cytoplasmic and nuclear chromatin ma-
turation is complete. The neutrophils and eosinophils have round, oval,
or bean-shaped nuclei in a mature cytoplasm. The cells may cause the
inexperienced technologist to report a leukogram indicating a left shift.
This condition has been reported and cats. 12 • 13 It seems possible that
these cells might have compromised abilities to migrate through vessel
walls due to an apparent rigidity of the nucleus. It has not been reported
that people and dogs with Pelger-Hiiet anomaly are more susceptible to
infection, however.
Chediak-Higashi syndrome has been found in cats as well as man,
mink, cattle, mice, and a killer whale (Fig. 8). 20• 26 Hematologically it is
characterized by enlarged neutrophilic and eosinophilic granules. Other
granules containing cells such as melanocytes have abnormal granules
also. Animals with Chediak-Higashi syndrome are partial albinos and
have photophobia and increased susceptibility to infection. The latter
may be due either to mechanical interference by the large granules
during neutrophil migration, 19 or to delayed delivery of the contents of
the enlarged lysosomal granules into phagosomes. 19• 26
252 JosEPH G. ZrNKL

Figure 7. Pelger-Huet anomaly in a neutrophil and an eosinophil of a dog. The

cytoplasm and the nuclear condensation are similar to those of normal cells, but the
nucleus is unsegmented. Oval and round nuclei may also be found . (l400 X)

Figure 8. Ch ediak-Higashi syndrome in a cat. The eosinophil granu les are la r ge

and round rather th an needle shaped. (l400 x ) (Courtesy of Dr. John Kramer, Washing-
ton State Unive rsity.)
L EU KO CYTES 253
Eosinophils
Gray-staining granules may be found in greyhound eosinophils. 35
Occasionally eosinophils with numerous vacuoles or with a few greatly
enlarged granu les are found in dog blood smears. A counterpart to
Charcot-Leyden crystals seen in humans with granu locytic leukemia is
not recognized in animals.
Monocytes
Morphologic changes in monocytes occasionally occur. Cells that
are probably promonocytes have a less ameboid nucleus and bluer and
scantier cytoplasm than normal monocytes. Such cells may be seen in
earlier stages of infectious diseases. Generally when these young cells
are seen, it can be anticipated that a moderate to m arked monocytosis
will follow.
Monocytes with characteristics of macrophages, that is, very abun-
dant and foamy cytoplasm, are occasionally found in blood smears (Fig.
9). Th eir significance is unknown.

Lymphocytes
Circulating "atypical" lymphocytes are seen in a number of nonleu-
kemic conditions. These cells may be undergoing blast transform ation
in response to antigen stimulation and are morphologically heteroge-
nous. They demonstrate a spectrum of increased size, nuclear folding,

Figure 9. Large , foamy mo nocyte of a dog. The abundant, foamy cytoplasm suggests
·. ~a t cells such as this are transformin g to macrophages in th e blood. (1400 x )
254 JosEPH G. ZrNKL

Figure 10. "Atypical" lymphocyte (left) of a dog. This cell contains basophil cyto-
plasm and an immature nucleus with a folded membrane. Better terms for this cell are
immunocyte or stimulated lymphocyte. (1400x)

immaturity of nuclear chromatin, prominence of cytoplasmic basophilia,

and prominence of cytoplasmic azurophilic granules (Fig. 10).38 Anum-
ber of conditions and agents including postimmunization, viral diseases,
autoimmune disease, salmon poisoning, and canine infectious hepatitis
may cause these cells to become prominent in the blood.
Occasionally plasma cells or plasma cell-like lymphoid cells, which
may represent a more advanced form of the atypical lymphocyte, are
seen in blood. Even Mott cells (that' is, plasma cells with large, clear
cytoplasmic pockets) are occasionally found in blood smears. Their sig-
nificance is unclear.

LEUKOCYTES IN DISEASE

Many diseases cause changes in leukocyte numbers or morphology.

Because leukocytes respond to diseases in limited but variable ways, the
changes are often of limited value in making specific diagnoses. How-
ever, they do provide information on the type of disease that is occur-
ring, the ability of an animal to respond to a disease agent, and the
changing character of the disease.
There are some diseases in which changes in leukocyte morphology
provide evidence specific enough to identify the disease. This may occur
frequently (as with salmon poisoning) or rarely (as with canine distem-
per).
LEUKOCYTES 255
Finally, there are diseases of the leukocytes themselves. These may
cause functional inabilities that make the animal susceptible to infec-
tions. These may be very difficult to recognize unless detailed investi-
gations are conducted.
Stress
There are a number of patterns of leukocyte responses to physio-
logic and pathologic conditions. Stress causes one of the most common
changes found in the leukogram of dogs and cats. The leukogram of
stress may be complicated by changes induced by other agents such as
bacteria and drugs. In the dog the stress reaction is characterized by
neutrophilia without a left shift, lymphopenia, monocytosis, and eosino-
penia. The neutrophil count is usually greater than 15,000 per pJ, but
occasionally it may be nearly 30,000 per ILL If a left shift of more than
a few (about 2 per cent) band neutrophils is present, other causes of
neutrophilia should be considered.
Lymphocyte counts are usually less than 1,500 per ILl in stressed
dogs; frequently the count is less than 1,000 per ILL In young dogs
significant lymphopenia occurs when the count is below 2,000 per ILL
Monocytosis of stress is usually mild, with counts in the high normal
range or slightly above the normal range. Eosinopenia occurs in stress.
Blood eosinophil counts also decrease in response to inflammation in-
dependent of their response to steroids; 3 therefore, eosinopenia is not
helpful in differentiating inflammation from stress-induced changes in
the leukogram.
Hematologically, cats respond to stress similarly to dogs except that
monocytosis usually does not occur. Significant lymphopenia occurs at
higher counts in cats than in dogs.
Physiologic Leukocyt~sis

Increased heart rate and splenic contraction due to excitement or

muscular activity can dislodge blood cells in sluggish areas of blood flow
and cause the spleen to release blood. These actions result in a rapid
increase in leukocytes, especially neutrophils and lymphocytes, in the
large vessels. Cats are more likely to develop physiologic leukocytosis
than are dogs. In both species the leukocyte count is in the high normal
range. The change is usually temporary, and resampling several hours
later often gives more normal results.
Inflammatory Disease
In dogs the most common response to disease is caused by inflam-
mation. The virulence of the inflammatory agent can often be judged
by the leukogram. Mild inflammation produces a leukocyte response
that is similar to the stress reaction. As organisms cause a more purulent
reaction, the neutrophil count increases markedly and a left shift, which
can include metamyelocytes and myelocytes, is found. In some condi-
256 JosEPH G. ZrNKL

tions, such as pyometra, the leukocyte count may exceed 100,000 per
JLI. Generally, localized infectious lesions elicit a much greater neutro-
philia than do agents that produce widespread inflammation. Perhaps
this is because in widespread inflammation a much greater egress from
the blood to the tissues occurs.
Massive infections or infections by an organism that produces a
potent toxin are usually associated with low or normal neutrophil count
and a marked left shift. The pathogenesis of this degenerative left shift
is due to extreme migration of cells into the tissues, or detrimental
effects of the toxins on granulopoiesis, or both. The left shift occurs in
an attempt to meet peripheral needs. Often the left shift is accompanied
by toxic changes. Unfortunately toxic cells and immature granulocytes
are not as functional as normal, mature neutrophils. Severe intestinal
diseases often cause a degenerative left shift. Massive leukocyte loss into
the gut occurs, and toxic bacterial products adversely affect maturation
and production of neutrophils. Feline infectious enteritis (panleuko-
penia) and canine parvovirus enteritis cause leukopenia in this manner.
Leukopenia is common in cats. The most obvious cause is infectious
enteritis. Total leukocyte counts may be less than 500 per JLI. Neutro-
phils are the most depressed cell type, but all leukocytes are decreased.
Differential counts may indicate a relative lymphocytosis (70 to 80 per
cent), but the absolute lymphocyte count is decreased. During recovery
from infectious enteritis, the leukogram is initially characterized by a
marked toxic, degenerative left shift. Early recovery is marked by rising
lymphocyte counts. Further into convalescence the left shift is less
marked and the cell count may be high.
Occasionally cats with feline infectious peritonitis have leukocyte
counts below 1,000 per JLI. In contrast to infectious enteritis, neutrophils
are the predominant white blood cell.·
Leukopenia in cats may occur in conditions other than infectious
enteritis. Among the many manifestations of FeLV infection are various
cytopenias, including leukopenia. FeLV-associated leukopenia may be
due to effects on the marrow cells or it may be due to tumor occupying
the marrow.
Young dogs with parvovirus-induced enteritis may be leukopenic.
In contrast, coronavirus enteritis does not cause leukopenia. 23 Probably
this difference is due to the severity of the diseases. Since parvovirus
enteritis is usually more severe, intestinal bacterial toxins probably are
higher in blood and depress the marrow more than in coronavirus en-
teritis. The extent of the lesions may also be greater and more blood
cells egress.
Other Causes of Leukocyte Changes
A wide variety of drugs can cause granulocytopenia in humans. 15
Some of these are phenothiozines, sulfonamides, phenylbutazone, and
chloramphenicol. Drug-induced granulocytopenia is most often due to
LEUKOCYTES 257
adverse effects on the precursors, but increased peripheral destruction
of granulocytes can also occur through immune-mediated mechanisms.
In these cases the drug acts as a hapten on the cell surface, the immune
system forms antibody to the hapten-protein complex, and the antibody
initiates the destruction of the granulocytes.
Neutropenia and other cytopenias may occur in dogs given phen-
ylbutazone,34 the synthetic estrogen estradiol cyclopentylpropionate
(ECP), 32 or thiacetarsamide. 40 Neutropenia may occur in cats treated
with antibiotics such as chloramphenicol.
Other causes of neutropenia in humans includes hypersplenism,
neonatal isoimmunization, and lupus erythematosus.
Lymphopenia is a common hemogram finding, and stress is the
most frequent cause. Lymphopenia may be seen with lymphomas.
Nearly all animals undergoing chemotherapy for cancer are lympho-
penic. In addition these animals are often anemic, thrombocytopenic,
and neutropenic. Many of these same drugs are used to treat autoim-
mune diseases and lymphopenia is expected. In fact, it is the desired
effect.

DISEASES OF LEUKOCYTES

A number of diseases in which leukocytes are abnormal are rec-

ognized in humans. These include the immunodeficiency diseases as
well as the inherited neutrophil defects. Only a few of these conditions
have been recognized in animals. In addition to leukocyte diseases, there
is a group of diseases in which the leukocytic changes may be of such
specificity as to provide a definitive diagnosis.
Canine Cyclic Hematopoiesis (Neutropenia)
Periodic, diminished hematopoiesis causes the disease cyclic he-
matopoiesis (or cyclic neutropenia) in grey ColliesY Blood reflects the
nature of the disease by having periodic fluctuations in the number of
leukocytes (particularly neutrophils), reticulocytes, and platelets. The
cycles have a periodicity of 11 to 13 days. Since neutrophils have a short
life-span compared to platelets and erythrocytes, the cyclic nature of the
neutrophil counts is the most obvious hematologic abnormality.
The disease is inherited as an autosomal recessive trait and is fre-
quently lethal during early life. Death usually is due to infectious agents
that gain a foothold during a neutropenic stage. Older dogs become
increasingly dependent upon antibiotics for survival. Most dogs die be-
fore they are two years old. Recent studies suggest that therapy with
lithium carbonate (150 to 300 mg per day) may eliminate the cycles and
maintain neutrophil numbers near normal. 16 Similar studies in people
on cancer chemotherapy suggest that lithium may decrease the rate of
infections due to compromised host resistance. 22 Perhaps lithium-stim-
258 JOSEPH G. ZINKL

ulated granulopoiesis maintains neutrophil counts at levels that can ef-

fectively combat microorganisms.
Canine Granulocytopathy
Recently, a defect of granulocytes of Irish Setters has been de-
scribed. 27 • 28 The disease is transmitted as an autosomal recessive trait
and is characterized by recurrent, life-threatening bacterial infections.
Dogs usually die young. The neutrophils have an impaired ability to kill
bacteria, which is associated with reduced glucose oxidation by the hex-
ose monophosphate pathway. The disease does not seem to have an
exact counterpart in humans, although detailed studies have not been
conducted to precisely determine the biochemical lesion.
Chediak-Higashi Syndrome
Chediak-Higashi syndrome has been described in a previous sec-
tion. It should be emphasized that Chediak-Higashi syndrome is a dis-
ease of neutrophils that results in decreased host resistance.
Immunodeficiency Syndrome
Inherited defects of lymphocytes, resulting in immunodeficiency
such as combined immunodeficiency of Arabian foals, has not been de-
scribed in dogs or cats. Several cases of Pneumocystis pneumonia, a
condition found in immunosuppressed humans, have been described in
miniature Dachshunds. 7 These reports suggested that immunodefi-
ciency may occur in this breed.
Canine Ehrlichiosis
Canine ehrlichiosis or tropical pancytopenia caused by Ehrlichia
canis affects the production of blood cells. 4 • 29 • 37 It is characterized by
inclusion bodies in monocytes and other leukocytes. The disease occurs
in many areas of the United States, occasionally epizootically. Based
upon the severity of the clinical signs and on the particular leukocytes
that contain inclusion bodies, there apparently are several strains of the
organism. An important vector and reservoir for the organism is the
brown dog tick, Rhipicephalus sanguineus.
Clinical and hematologic manifestations vary with the stage of the
disease, the virulence of the organism, and the breed of dog affected. 5 • 29
Severe infection is characterized by high fever, anorexia, weight loss,
serous nasal and ocular discharges, lymphadenopathy, and splenomeg-
aly, beginning about 10 to 14 days after initial contact with the organism.
During this time pancytopenia may be evident, but hemorrhagic epi-
sodes do not occur. However the infection often persists. In some dogs
a chronic or delayed disease occurs after 50 to 100 days. Marked pan-
cytopenia develops, and hemorrhage, epistaxis, and edema occur; many
dogs die. The pancytopenia of this stage is probably due to destruction
of precursor cells in the marrow. Leukopenia increases the suceptibility
LEUKOCYTES 259
of those dogs to secondary infections. German Shepherd Dogs seem to
be particularly susceptible to the disease.
Hematologic examination reveals, in addition to anemia, leuko-
penia, severe thrombocytopenia, and leukocytes containing morula.
Most often these are in monocytes, but other leukocytes may be infected.
Hyperproteinemia is common.
A definitive diagnosis of ehrlichiosis is made by demonstrating the
organisms in leukocytes (Fig. 11 ) or by a positive indirect fluorescent
antibody test. In suspect cases, the leukocytes from several blood smears
may have to be examined. Buffy coat smears facilitate the leukocyte
examination.
Recently we have observed equine ehrlichiosis morulae in neutro-
phils of a dog being treated for lymphosarcoma. Presumably the dog
was immunosuppressed sufficiently to allow the organisms to become
established. The dog was from the Sierra Nevada foothills, an area that
is enzootic for equine ehrlichiosis. Blood from the dog produced ehr-
lichiosis in horses, and morulae were found in neutrophils and eosino-
phils of the horses.
Mucopo1ysaccharidosis
Lymphocytes, monocytes, and occasionally neutrophils containing
azurophilic granules may be seen in mucopolysaccharidosis, a rare con-
dition of dogs. 30 The granules are usually larger and more darkly stain-
ing than the granules found in a few lymphocytes of some dogs. Oc-
casionally a clear space surrounds the granule in lymphocytes, and the

Figure II. Ehrlichia canis inclusion in a lymphocyte. (l400 x )

260 JosEPH G. ZrNKL

granule-containing cells are usually numerous. When in neutrophils,

the granules stain reddish-brown. This condition has been reported to
also occur in cats.
Acute Infectious Lymphocytosis-like Syndrome
Stimulated ("atypical") lymphocytes, similar to those seen in dogs
undergoing an immunologic response, have been seen in young dogs.
The condition resembles, in part, acute infectious lymphocytosis of chil-
dren. Clinically the condition was characterized by mild fever and list-
lessness. Occasionally the dogs had diarrhea and vomiting. Peripheral
lymph nodes and the tonsils were usually enlarged. The blood lympho-
cytes were similar to those seen in infectious canine hepatitis. An attempt
at transmission was unsuccessful. 9
Infectious Canine Hepatitis
Early in the course of infectious canine hepatitis, dark-staining
mononuclear cells may be found. These cells are difficult to classify but
are probably stimulated lymphocytes. During convalescence they are
replaced by normallymphocytes. 33
Salmon Poisoning Disease
Dogs with salmon poisoning (a misnomer) often have blood smears
with stimulated lymphocytes. The disease is caused by Neorickettsia hel-
minthoeca, which is transmitted through the salmon fluke Nanophyetus
salmincola. The stimulated lymphocytes appear after an initial leuko-
penia. In addition to these cells, some dogs also have macrophage-like
monocytes in the blood. This cell has an abundant, highly vacuolated
cytoplasm. 31
When these cells, a recent history of.having eaten uncooked salmon,
and the clinical signs of fever, vomiting, diarrhea, and enlarged lymph
nodes are present, a presumptive diagnosis of salmon poisoning can be
made. Confirmation is reached by finding the fluke ova in the feces or
by observing characteristic changes in lymph node aspirates. Cytologi-
cally, the lymph node aspirate is classified as reactive. There is an in-
creased number of basophilic lymphoblasts and plasma cells. A rather
marked increase in monocytes is also seen. Many of these cells contain
amorphous debris. 41 Occasionally groups of very small, rod-shaped or-
ganisms are found in macrophages. These may be in a discrete, round
structure (elementary body) or they may completely fill the cell.
Canine Distemper
Canine distemper may be characterized by leukopenia, although
normal or elevated leukocyte counts have been found. Lymphopenia is
often seen, particularly in the early stages. On rare occasions leukocytes
and even erythrocytes may contain inclusion bodies, which are usually
pale basophilic, although slightly acidophilic bodies are also found. 35
LEUKOCYTES 261
The erythrocytes that contain inclusions are young cells, and many are
probably reticulocytes. Affected dogs usually have hard pads. 35

SUMMARY

Dogs and cats respond to many diseases by changes in leukocyte

numbers. Infectious diseases often cause leukocytosis due to neutro-
philia. Left shift may accompany the leukocytosis, indicating that the
marrow is mounting a response to the disease. Left shift also indicates
that the marrow has fallen somewhat behind the needs of the animal.
Degenerative left shift is considered a poor sign.
Lymphopenia and eosinopenia also are found in infectious diseases.
Lymphocytosis may occur during the recovery or if the disease becomes
chronic. The nature and duration of the infection determine the mag-
nitude of the monocyte response. It is erroneous to consider a disease
chronic based only upon monocytosis. Trauma, autoimmunity, or any
disease with significant tissue destruction can evoke a monocyte re-
sponse.
Leukopenias are relatively common in cats and are found with mod-
erate frequency in dogs. Drugs, viral diseases (such as FeL V, feline en-
teritis, parvovirus), ehrlichiosis, and hereditary conditions may cause
panleukopenias or single leukopenias.
Occasionally leukocyte examination provides evidence of a specific
etiology (such as with ehrlichiosis). Sometimes changes occur which, al-
though not specific for a disease, may provide a strong evidence of a
particular disease (such as in salmon poisoning).
Leukocyte evaluation should include not only total count and dif-
ferential count (with' calculation of absolute numbers of the different
cells) but also morphologic examination of the cells by qualified people.
In many practices the only qualified person is the veterinarian.

REFERENCES

I. Aethans, D., Keller, H. U., Hess, M. W., eta!.: Impaired neutrophillac0rnotion during
acute bacterial infections. Int. Arch. Allergy Appl. Immunol., 61:3'2'1, 1980.
2. Atkinson, K., Deeg, H.]., Storb, R., eta!.: Canine lymphocyte subpopulations. Exp.
Hematol., 8:821, 1980.
3. Bass, D. A., Goniva, T. A., Szejda, P., et a!.: Eosinopenia of acute infection.]. Clin.
Invest., 65:1265, 1980.
4. Beeson, P. B., and Bass, D. A.: The EosinophiL Philadelphia, W. B. Saunders Co.,
1977.
5. Buhles, W. C., Jr., Huxsoll, D. L., and Hildebrandt, P. K.: Tropical canine pancyto-
penia: Role of aplastic anemia in the pathogenesis of severe disease. J. Camp. Pathol.,
85:511, 1975.
6. Cline, M. J.: The White Cell. Cambridge, Mass., Harvard University Press, 1975.
7. Copland, ]. W.: Canine pneumonia caused by Pneumocystis carinii. Aust. Vet. J.,
50:515, 1974.
262 JOSEPH G. ZINKL

8. Craddock, C. G.: Production, distribution, and fate of granulocytes. In William, W.

J., Beutler, E., Frslev, A. J., eta!. (eds.): Hematology. New York, McGraw-Hill Book
Co., 1972.
9. Demartini, J.: Personal communication, 1980.
10. Dvorak, A.M., and Dvorak, H. F.: The basophil. Arch. Pathol. Lab. Med., 103:551,
1979.
11. Fehr, J., and Grossman, H-C.: Disparity between circulating and marginated neutro-
phils: Evidence from studies on the granulocyte alkaline phosphatase, a marker of
cell maturity. Am. J. Hematol., 7:369, 1979.
12. Feldman, B. F.: Personal communication, 1980.
13. Feldman, B. F., and Ramans, A. U.: The Pelger-Hiiet anomaly of granulocyte leu-
kocytes in the dog. Canine Pract., 3 :22, Oct. 1976.
14. Finch, S.C.: Granulocytosis. In Williams, W. J. Beutler, E., Erslev, A. J., eta!. (eds.):
Hematology. New York, McGraw-Hill Book Co., 1972.
15. Finch, S. C.: Granulocytopenia. In Williams, W. J., Beutler, E., Erslev, A. J., et a!.
(eds.): Hematology. New York, McGraw-Hill Book Co., 1972.
16. Hammond, W. P., and Dale, D. C.: Lithium therapy of canine cyclic hematopoiesis.
Blood, 55:26, 1980.
17. Hughes, W. T.: Pneumocystis carinii pneumonia. N. Engl. J. Med., 297:1381, 1977.
18. Jamuar, M. P., and Cronkite, E. P.: The fate of blood granulocytes. Exp. Hematol.,
8:884, 1980.
19. Klebanoff, S. J., and Clark, R. A.: The Neutrophil: Function and Clinical Disorders.
New York, North-Holland Publishing Co., 1978.
20. Kramer, T. W., Davis, W. C., and Prieur, D. J.: The Chediak-Higashi syndrome of
cats. Lab. Invest.,36:554, 1977.
21. Lund, J. E., Padgett, G. A., and Ott, R. L.: Cyclic neutropenia in grey collie dogs.
Blood, 29:452, 1967.
22. Lyman, G. H., Williams, C. C., and Preston, D.: The use of lithium carbonate to
reduce infection and leukopenia during systemic chemotherapy. N. Engl. J. Med.,
302:257, 1980.
23. Moreau, P. M.: Canine viral enteritis. Com pend. Contin. Ed., II :540, 1980.
24. Murphy, P.: The Neutrophil. New York, Plenum Medical Book Co., 1976.
25. Park, B. Y., and Good, R. A.: Principles of Modern Immunology. Philadelphia, Lea
and Febiger, 1974.
26. Prieur, D.J, and Collier, L. L.: Chediak-Higashi syndrome of animals. Am.J. Pathol.,
90:533, 1978.
27. Renshaw, H. W., and Davis, W. C.: Canine granulocytopathy syndrome: An inherited
disorder of leukocyte function. Am. J. Pathol., 95:731, 1979.
28. Renshaw, H. W., Davis, W. C., and Renshaw, S. T.: Canine granulocytopathy syn-
drome: Defective bactericidal capacity of neutrophils from a dog with recurrent
infections. Clin. Immunol. Immunopathol.,8:385, 1977.
29. Ristic, P. I., and Small, E.: Canine ehrlichiosis: The role of the spleen in protection
against the disease. Vet. Med./Small Anim. Clin., 74:1469, 1979.
30. Schalm, 0. W.: Mucopolysaccharidosis. Canine Pract., 4:30, Dec. 1977.
31. Schalm, 0. W.: Leukocyte counts and lymph node cytology in salmon poisoning of
dogs. Canine Pract., 5:59, June 1978.
32. Schalm, 0. W.: Ex~ous estrogen toxicity in the dog. Canme Pract., 5:57, Oct.
1978.
33. Schalm, 0. W.: Special characteristics of 'lymphocytes and monocytes in infectious
canine hepatitis. Canine Pract., 6:51, Jan. 1979.
34. Schalm, 0. W ..: if>ibleny1bntazone toxicity in two dogs. Canime Pract., 6:47, July 1979.
35. Schalm, 0. W .. , Jain, N. C., and Carroll, E. J.: Veterinary Hematology. Edition 3.
Philadelphia, Lea and Febiger, 1975.
36. Schiffmann, E., Corcaran, B. A., and Wahl, S. M.: N-Formylmethionyl peptides as
chemoattractantsforleukocytes. Proc. Nat!. Acad. Sci. USA, 72:1059, 1975.
37. Seamer, T., and Snape, T.: Ehrlichia canis .and tropical canine pancytopenia. Res.
Vet. Sci., I3:307, 1972.
38. Shiftan, T. A., and Mendelsohn, J.: The circulating "atypical" lymphocyte. Hum.
Pathol., 9:51, 1978.
LEUKOCYTES 263
39. Showell, H.]., Freer, R. ]., Zigmond, S. H., et al.: The structure-activity relationships
of synthetic peptides as chemotactic factors and inducers of lysosomal enzyme se-
cretion for neutrophils.]. Exp. Med., 143:1154, 1976.
40. Watson, A. D.].: Bone marrow failure in a dog. J. Small Anim. Pract., 20:681, 1979.
41. Zinkl, J. G., and Keeton, K. S.: Lymph node cytology. II. Calif. Vet., 33:6, April1979.

Department of Clinical Pathology

School of Veterinary Medicine
University of California, Davis
Davis, California 95616