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Comparative Morphology, Development, and

Function of Blood Cells in Nonmammalian
Vertebrates

Article in Journal of Exotic Pet Medicine · April 2009

DOI: 10.1053/j.jepm.2009.04.006

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Juan Alberto Claver Agustín Quaglia

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Topics in Medicine and Surgery

Comparative Morphology, Development, and

Function of Blood Cells in
Nonmammalian Vertebrates
Juan A. Claver, DVM, PhD,
and Agustin I. E. Quaglia

Abstract
Much of our knowledge regarding vertebrate blood and blood cells is based on
mammalian references. The critical study of nonmammalian vertebrate blood is
relatively new, and comparatively few investigations have been published that
focus on these animals’ ontogeny and structure-function relationships of blood
cells. Nonmammalian vertebrates comprise birds, reptiles, amphibians, and
fishes, all of which have a wide range of forms and adaptations. For the
clinician, a lack of basic knowledge with these species makes the evaluation of
a hemogram more challenging than in mammals. This is a concise review of our
current knowledge of comparative morphology that describes routine staining
procedures and the development and function of blood cells in nonmammalian
vertebrates. Copyright 2009 Elsevier Inc. All rights reserved.

Key words: amphibian; bird; fish; hematology; reptile

M
any general considerations can be made
for the blood of nonmammalian verte-
brates (NMV). As in mammals, NMV blood
cells include erythrocytes, leukocytes, and hemo-
static cells. With very few exceptions, red blood cells
in NMV are nucleated and oval in shape.1 A cytoskel-
eton is responsible for their conversion from a
sphere to an ellipse, and imparts reversibility of trau-
matic deformation. All NMV have nucleated throm-
bocytes instead of platelets, and heterophils with
acidophilic granules are, in most cases, the counter-
part of mammalian neutrophils.
Birds
Birds include more than 9000 species of homeother-
mic vertebrates. Most of the scientific information
regarding avian hematology is the result of domestic
fowl investigations. The morphology of avian blood
cells are comparatively more homogeneous between
orders than in other NMV.
Hematopoiesis
Current scientific knowledge regarding early hema-
topoiesis in vertebrates came from experiments first
performed on the avian embryo. Primitive hemato-
poiesis in the chicken embryo begins in the blood
From the Histology and Embryology Department, Facultad de
Ciencias Veterinarias, Universidad de Buenos Aires, Buenos
Aires, Argentina, and Fundación de Historia Natural Félix de
Azara, Buenos Aires, Argentina.
Address correspondence to: Juan A. Claver, DVM, PhD, His-
tology and Embryology Department, Facultad de Ciencias Veteri-
narias, Universidad de Buenos Aires, Buenos Aires, Argentina.
E-mail: jclaver@fvet.uba.ar.
© 2009 Elsevier Inc. All rights reserved.
1557-5063/09/1802-$30.00
doi:10.1053/j.jepm.2009.04.006

Journal of Exotic Pet Medicine, Vol 18, No 2 (April), 2009: pp 87–97 87

88
Figure 1. Avian blood cells (A, B, C). Erythrocyte (a), polychromatic erythrocyte (b), thrombocytes (c), heterophil (d), eosinophil (e), basophil
(f), small lymphocyte (g), large lymphocyte (h), and monocyte (i).
islands of the yolk sac during the second day of
incubation. At that time, only nucleated erythrocytes
and thrombocytes are produced.2,3 Hemoglobin can
be detected as early as 24 hours after fertilization
(2-4 somite stage). The yolk sac in avian embryos will
remain hematopoietic for most of the embryonic
development, until bone marrow hematopoiesis is
established.3,4 Definitive hematopoiesis begins in the
dorsal mesenchyma (aortic and para-aortic foci)
from days 5 to 8 of incubation. New stem cells from
these areas colonize the spleen, which is hematopoi-
etic from days 9 to 18, and finally the bone marrow
from day 12 of incubation into adult life. The fetal
liver is not a major hematopoietic organ in birds.
Claver and Quaglia
Birds were an early model for immunology studies,
leading to the identification of the B- and T-lympho-
cyte lineages. Maturation of T-lymphocytes occurs in
the thymus. Birds have a specific organ, the bursa of
Fabricius, which is the site of differentiation for B-
lymphocytes.2
Erythrocytes
Avian erythrocytes are oval in shape and nucleated
(Fig 1A, a). The nucleus of the avian erythrocyte is
also oval in shape, and it becomes more condensed
with age. The cytoplasm generally stains uniformly
orange-pink, except for a thin, pale perinuclear
Morphology of Blood Cells in Nonmammalian Vertebrates
band. Hemoglobin is found in both the cytoplasm
and nucleus. Erythroplastids (anucleated erythro-
cytes) and immature erythrocytes are occasionally
present in circulating blood. Immature forms of the
avian erythrocytes (Fig 1A, b) have an irregular cy-
toplasmic polychromasia and a more rounded, pale
nucleus.5-7 The major function of erythrocytes is ox-
ygen transport, but recently it has been reported that
bird erythrocytes, as nonimmune cells, are able to
participate in some immune responses that contrib-
ute to host defense.8
Leukocytes
In the majority of birds, heterophils are the most
common white blood cell in circulation. Heterophils
have colorless cytoplasm and typical eosinophilic,
rod-shaped granules, together with some other
rounded granules (Fig 1A, d). Specific granules are
elliptical, although in some species they may be oval
or rounded and have a distinct central body that
appears refractile.5 Heterophilic response to infec-
tion is similar to that of the mammalian neutrophil,
migrating to the sites of inflammation and killing
pathogens.9 Heterophils are highly phagocytic and
are capable of a broad spectrum of antimicrobial
activity. These leukocytes tend to accumulate in in-
flamed tissue, causing tissue damage and forming
heterophilic foci that are morphologically similar to
inflammatory lesions in reptiles.10,11 The avian het-
erophils contain many lysosomal and nonlysosomal
enzymes.12 Unlike mammalian neutrophils, avian
heterophils are devoid of myeloperoxidase and de-
pend primarily on nonoxidative mechanisms, ly-
sozyme, and acid phosphatase for antimicrobial ac-
tivity.13 The B defenses have a broad spectrum of
activity against the binding abilities of infectious
agents. Spherical granules contain acid hydrolases,
whereas a third, smaller, round vacuole granule has
been identified by electron microscopy.12 Het-
erophils may show toxic changes in severe systemic
illness, including increased cytoplasmic basophilia,
degranulation, and abnormal granulation.5
Eosinophils are similar in size to heterophils but
contain round, eosinophilic granules and a pale blue
cytoplasm (Fig 1A, e).5 In some bird groups, the
granules may actually be rod shaped (e.g., Anatidae,
Tinamidae, Falconidae; Fig 1D), whereas in others
they may have a bluish coloration (e.g., in psittacine
birds; Fig 1B and C).14 Crystalline cores have been
described in specific granules of eosinophils in some
birds such as geese and ducks, but these structures
are not commonly found in other species.15 Cyto-
chemically, avian heterophils lack myeloperoxidase,
whereas eosinophils are peroxidase positive.13 Addi-
89
tionally, certain hydrolytic enzymes (e.g., acid phos-
phatase and arylsulphatase) have been detected in
avian eosinophilic granules, supporting the theory
that these structures are lysosomal in nature. The
function of the avian eosinophil is poorly under-
stood. Eosinophilia can be observed after foreign
antigen administration and possibly in association
with alimentary tract parasitism, but the parasitic
response by this cell type in birds has not been
scientifically confirmed.11,16
Basophils are recognized by a rounded nucleus
and characteristic violet to reddish purple granules,
which are much smaller than those of the eosinophil
(Fig 1A, f). Little is known about basophil function
in birds. Basophils appear to play an important role
in early inflammation and immediate hypersensitiv-
ity reactions, but differ from those in mammals by
not contributing to delayed hypersensitivity.17 Severe
stress has also been proposed as an underlying cause
for an increased basophilic response in birds.18
Lymphocytes may be the major circulating leuko-
cyte in the blood of some species. Avian lymphocytes
are small to large, rounded to irregular cells with a
round nucleus and scant to abundant basophilic
cytoplasm (Fig 1A, g and f). Small lymphocytes pre-
dominate in most birds, with irregular projections or
blebs frequently observed on this cell type.7
Birds were used as a pioneer model to study im-
munology, which resulted in the identification of the
B- and T-lymphocyte lineages. Avian lymphocytes
appear to function in the same manner as mamma-
lian lymphocytes. The B-cells (bursa dependent)
have immunoglobulin receptors for antigens, while
T-cells (thymus dependent) are involved in cell-me-
diated immunity.2,7
Monocytes are the largest cells in bird blood, and
they resemble their mammalian counterparts. The typ-
ical monocyte is round, with a kidney-shaped nucleus.
The cytoplasm is generally deep blue or grayish blue,
often presenting a pink- or purple-staining granular
area near the nucleus (Fig 1A, i). Monocytes have more
cytoplasm and a paler nucleus than large lymphocytes.7
As in mammals, avian monocytes are part of the mono-
cyte-macrophage system and constitute the replace-
ment pool for tissue macrophages.16
Thrombocytes
In all NMV, thrombocytes are the functional equivalent
of mammalian platelets. In birds, thrombocytes are
round to oval cells, smaller than erythrocytes, and con-
tain an oval to rounded nucleus. The cytoplasm is light
blue or colorless, often vacuolated with a few acido-
philic granulations (Fig 1A, c).7 Thrombocytes are of-
ten confused with small lymphocytes and have a ten-
90
dency to clump in smears, particularly when processed
without the use of an anticoagulant. When clumping
occurs, they may show degranulation of specific gran-
ules, cellular degeneration, and nuclear pyknosis.16 As
with mammalian platelets, bird thrombocytes aggre-
gate in a site of vascular injury and form a hemostatic
plug. Additionally, thrombocytes have phagocytic abil-
ities and probably have some function in nonspecific
immunity.19,20 Thrombocyte abnormalities are rarely
diagnosed and thrombocyte dysfunction syndromes
are not described in birds.
Reptiles
Reptiles comprise nearly 8000 species of poikilother-
mic vertebrates. Reptiles represent a diverse group
of animals (e.g., lizards, snakes, turtles, crocodiles),
with some of them used as pets and others as an
economic resource. The American anole, Anolis caro-
linensis, is the “laboratory mouse” of the reptile world
in terms of research investigation. The basic hema-
tology of reptiles is very similar to that of birds,
although reptiles are a more heterogeneous class so
it may be more difficult to draw inferences between
species.21
Hematopoiesis
Few studies have been made in primitive hematopoi-
esis in reptiles. The yolk sac blood islands seem to be
the unique primary erythropoietic site during em-
bryonic life in reptiles, with bone marrow not be-
coming active until birth.22 As in birds, postnatal
erythropoiesis and granulopoiesis of reptiles occurs
mainly in the bone marrow, although the liver and
spleen have some hematopoietic function in the
early stages of development. Lymphocytes most
likely of bone marrow origin will colonize and dif-
ferentiate into T-cells in the thymus, which regress
with age. A true bursa of Fabricius has not been
found in reptiles, and the site where B-lymphocytes
develop is currently unknown. The location of lym-
phopoiesis includes the bone marrow, spleen, and
other locations within the body. Mucosal-associated
lymphoid tissue is also present in the digestive and
respiratory tracts.23
Erythrocytes
Erythrocytes of reptiles are similar in function and
appearance to those of birds but can vary in size,
being relatively smaller in Sauria and larger in some
Chelonia, Crocodilia, and Rhynchocephalia (Fig 2A,
a and b). The overall number of circulating erythro-
cytes is lower in reptiles than in mammals or birds.
Claver and Quaglia
Lizards generally have a greater erythrocyte popula-
tion than do snakes. There appears to be an inverse
correlation between erythrocyte number and cell
size.24,25 The nucleus of the reptile erythrocyte is
more rounded, particularly in turtles, and often has
irregular margins. Immature erythroid forms have
basophilic cytoplasm, similar to those in avian spe-
cies (Fig 2A, c). As reptiles recover from brumation,
there may be a marked regenerative response with
basophilic erythrocytes.26 The life of a reptile eryth-
rocyte has been reported to be 600 to 800 days, with
the extreme longevity being attributed to the low
metabolic rate of these animals.27
Leukocytes
Classification schemes for leukocytes in reptiles are
inconsistent because variable criteria have been used
to categorize these cells. Reptile leukocytes have
been described according to their appearance rather
than function. As in birds, reptiles have heterophils
instead of neutrophils. Heterophils may vary greatly
between groups, genera, and species. The reptile
heterophil has characteristic needle-like acidophilic
cytoplasmic granules (Fig 2A, g and B), with croc-
odilian species having larger granules, although
fewer in number, than lizards and snakes (Fig 2A,
f).28 The heterophil nucleus has a round shape, and it
may have two lobes in some lizards.26 Functionally, the
reptilian heterophil is similar to the avian heterophil,
with their primary function being phagocytosis.23
Azurophils are leukocytes similar to monocytes but
contain prominent azurophilic granules (Fig 2A, i).
Azurophils have been described in Squamata (snakes
and lizards) and Crocodilia, but are seen rarely in
Chelonia.11,29 Some authors consider azurophils as a
monocyte variant.30
Eosinophils, Basophils, Lymphocytes, Monocytes.
Eosinophils are observed in Crocodilia and Chelo-
nia, but their existence in Squamata is controversial.
Even inside a genus of snakes, eosinophils are de-
scribed in some species and not in others.31,32 Eosin-
ophils have a round nucleus and acidophilic-stain-
ing granules, although they stain blue in green
iguanas (Fig 2A, h). Cytochemically, heterophils
of both avian and reptilian species studied lack
myeloperoxidase, whereas eosinophils were perox-
idase positive.23
Basophils found in reptile species resemble those of
birds, both in appearance and function (Fig 2a, I).
Chelonians tend to have higher numbers of circulating
basophils than other groups of reptiles; in some cases
up to 50% to 60% of their differential may be attrib-
Morphology of Blood Cells in Nonmammalian Vertebrates 91

Figure 2. A, Blood cells of reptiles. Erythrocytes of crocodile (a) and iguana (b), polychromatic erythrocyte of turtle (c), normal (d) and reactive
(e) thrombocytes, heterophils of crocodile (f) and turtle (g), eosinophil of turtle (h), azurophil (i), monocyte (j), small lymphocyte (k), and basophil
(l). B, Heterophil. Trachemys scripta dorbigny.

uted to basophils. It is not known why chelonians have Amphibians

such high levels of circulating basophils.
For most reptiles, lymphocytes are the most preva-
lent circulating cell. They are similar in appearance to
the avian and mammalian lymphocytes (Fig 2A, k). B-
and T-lymphocytes have been characterized in rep-
tiles and are functionally similar to those found in
birds.23 Lymphocytes show seasonal variations in rep-
tiles, with lower numbers reported during the winter
months.33
Monocytes (Fig 2A, j) and thrombocytes (Fig 2A,
d and e) are similar in structure and function to
those described in the previous avian section.23
Very little has been published regarding amphibian
(class Amphibia) hematology. This class comprises
near 6000 species of poikilothermic vertebrates.
Frogs and other amphibians have been widely main-
tained in captivity as pets and as animal models for
physiologic and embryologic studies. Xenopus laevis,
the African clawed frog, is the most popular amphib-
ian species used in research. Amphibians are also
marketed as culinary delicacies and increasingly
studied as bio-indicators of environmental health.
The examination of blood samples can be useful in
92
evaluating the status of diseased animals, although
the similarity in function of amphibian blood cell
types and those of other species is largely unknown.
Hematopoiesis
The development of hematopoiesis in amphibians
has some similarity to that of birds and mammals.
The ventral blood islands (e.g., analogous to the
yolk sac blood islands of higher vertebrates) and
the dorsal lateral plate region are the sites of
primitive and definitive hematopoiesis in the am-
phibian embryo. Hematopoietic stem cells of the
dorsal lateral plate give rise to the definitive lin-
eages, which ultimately colonize the fetal liver and
thymus.34 The kidneys are the main sites of blood
formation in the larval stages of amphibian spe-
cies. The thymus is the major site of T-cell matu-
ration, but the site of B-cell lymphopoiesis is less
clear and amphibians appear to use different sites
for this specific cell production depending on the
species in question.35 Erythrocyte production,
along with thrombocytopoiesis, in adult urodelans
is conducted intravascularly in the spleen. In an-
urans, the spleen is usually the major site of eryth-
rocyte production, although the liver also serves as
a secondary locus for this activity. Bone marrow
hematopoiesis makes its phylogenetic debut in am-
phibians and typically occurs after metamorphosis
or hibernation. The bone marrow is hematopoiet-
ically active in some salamanders and Rana spe-
cies, but not in Xenopus.36,37 Seasonal changes can
cause a shift in the sites (e.g., spleen and bone
marrow) of hematopoiesis in some amphibians.38
Erythrocytes
Amphibians have nucleated, oval, flattened, bicon-
vex erythrocytes (Fig 3A, a and b). The amphibians
have the largest erythrocytes of the animal kingdom,
with Amphiuma tridactilum being recorded as having
the largest erythrocytes (70 ⫻ 40 ␮m). Anuran eryth-
rocytes are generally much smaller (22 ⫻ 15 ␮m)
than those reported for the urodelans. Conversely,
the erythrocyte count and hemoglobin concentra-
tion of these species are low.38 Although amphibians
have nucleated erythrocytes, a percentage of anucle-
ated erythrocytes (erythroplastids) may be observed
in salamanders (family Plethodontidae), with Batra-
choseps attenuatus possessing the record of nearly
95%.39 Maturation of the erythrocytes in the circula-
tion is common (e.g., Urodeles).38,40 As in reptiles,
the amphibian erythrocyte lifespan is longer than
that recorded for mammals and birds (e.g., about
700-1400 days in Bufo marinus).27 During brumation,
there is an increase in the lifespan of the peripheral
Claver and Quaglia
blood cells as a consequence of a reduced metabolic
activity and a slowing of hematopoiesis.41
Leukocytes, Eosinophils, Basophils,
Lymphocytes, Monocytes, Thrombocytes
Amphibian leukocytes are larger (30-32 ␮m) than
those described for other NMV. Although the pri-
mary acute inflammatory cell of the amphibian is the
heterophil, these animals also have a small number
of cells that have the staining characteristics of neu-
trophils (Fig 3A, d and e).42 Little is known about the
characteristics of these “neutrophils.” Heterophils
have acidophilic rod-shaped granulations and a seg-
mented nucleus (Fig 3B). During the hibernation
phase of frogs, an increase in the nuclear segmenta-
tion of neutrophils has been observed, probably due
to cell aging. Amphibian heterophils do phagocytize
bacteria.42
Amphibian eosinophils are similar to their het-
erophils, although the cytoplasmic granules are
larger, round to oval, and the nucleus is less seg-
mented (Fig 3A, f).42 Histochemically, eosinophils
are peroxidase negative and heterophils are peroxi-
dase positive.37 Little is known of eosinophil function
in amphibians, although some reaction to trematode
infestations has been documented.43
Basophils are morphologically similar to those of
other NMV and may be in low or high numbers
depending on the species (Fig 3A, g). Heparin-like
substances have been described in the basophils of
some amphibian species, and they appear to have a
similar function to basophils found in other NMV.43
Azurophils have been described in some amphib-
ian species and are believed to be a form of mono-
cyte or neutrophil (Fig 3A, j).42,43
Lymphocytes are generally the smallest leukocyte in
amphibians, being half the size of granulocytes. These
cells are round to ovoid in shape and contain a small
amount of cytoplasm (Fig 3A, h).42 Amphibian lym-
phocytes seem to act in specific immunity, although
urodelans do appear to lack some of the more evolved
aspects of lymphocyte-mediated specific immunity
present in anurans, birds, and mammals.44
Amphibian monocytes can vary in size, with some
being smaller and others being larger than the gran-
ulocytes (Fig 3A, i). Monocytes generally have a
higher cytoplasmic to nuclear ratio than lympho-
cytes and a rounded, kidney-shaped or horseshoe-
shaped nucleus. It can be difficult to differentiate
monocytes from large lymphocytes in some cases.42
As with other vertebrates, monocytes serve as phago-
cytic and antigen-presenting cells.37
Thrombocytes of amphibians are similar in structure
and function to those of birds and reptiles (Fig 3A, c).42
Morphology of Blood Cells in Nonmammalian Vertebrates 93

Figure 3. A, Blood cells of amphibians. Erythrocyte (a), polychromatic erythrocyte (b), thrombocytes (c), neutrophil (d), heterophil (e), eosinophil
(f), basophil (g), small lymphocyte (h), monocyte (i), and azurophil (j). B, Heterophils. Buffo arenarum (Anura). Courtesy of Mariana Cabagna.

Fishes
Fishes are the most numerous (27,000 species) and
diverse of all vertebrate groups, making generaliza-
tions regarding hematology difficult, if not impossi-
ble. The lampreys and the hagfish include about 100
species of jawless fishes. There are also 600 species of
chondrichthyes and more than 26,000 bony fishes
(osteichthyes).
Hematopoiesis
All fish lack bone marrow and lymph nodes.45 Elasmo-
branchs possess a thymus and spleen, but in the ab-
sence of bone marrow and lymph nodes, these fish
have evolved unique lymphomyeloid tissues, namely
epigonal and Leydig’s organs, associated with the
esophagus and gonads, respectively. The epigonal and
Leydig’s organs are thought to be vital in generating
the immune response in chondrichthyan fishes.45
94
Bony fishes (teleosts) form embryonic erythroid
cells in a distinct dorsal-lateral compartment of the
embryo known as the intermediate cell mass of
Oellacher and are, therefore, an exception to the
general vertebrate rule of embryonic hematopoiesis
on the yolk sac. Both primitive and definitive cells
arise from these cellular precursors.4
In many of the bony fishes, the renal interstitium is
the primary site of definitive hematopoiesis, with a
significant number of species possessing a discrete an-
terior (pronephros) kidney that is entirely devoted to
hematopoiesis. In the kidney interstitium, all lines of
hematopoietic differentiation are observed, including
erythropoiesis, granulopoiesis, and lymphopoiesis.46 In
some species of bony fishes, other organs (e.g., spleen,
heart, meninges, spiral valve) may have lymphomy-
eloid activity.47
Erythrocytes
Almost all fish have nucleated erythrocytes (Fig 4,
a). Remarkable exceptions are Maurolicus müelleri,
a teleost with small anucleated erythrocytes, and
the Antarctic ice fishes (family Channichthyidae),
which do not have erythrocytes or hemoglobin in
Figure 4. Blood cells of fishes. Erythrocyte (a), normal (b) and activated (c) thrombocytes, neutrophil of trout (d), heterophil of shark (e),
eosinophil (f), monocyte (g), basophil (h), and lymphocyte (i).
Claver and Quaglia
the blood. Erythrocytes in fishes are generally oval
in shape with an oval to rounded nucleus.48 The
elongated fish erythrocytes appear to represent
more mature cells.49 Erythrocytes are rounded in
some Agnathae (family Petromyzontidae)50 and
tend to be more rounded in elasmobranchs than
in teleosts. Fish blood has characteristically low
concentrations of erythrocytes (1 to 5 ⫻ 106/mm3)
and hemoglobin. A continued increase in size and
hemoglobin content occurs in circulating erythro-
cytes during their lifespan.51 Sedentary fishes have
lower erythrocyte levels than those in more active
fishes. Recently, it has been observed that, as in
birds, fish erythrocytes can act as immune cells,
binding and engulfing Candida albicans.52
Leukocytes
In fishes, the heterophil has been variably called
heterophil or neutrophil depending on the size of
cytoplasmic granules. In some instances, fish het-
erophils are referred to as acidophils.45 To avoid
confusion, certain authors propose to use the term
“neutrophil” only for mammals.26 Unlike avian and
reptilian heterophils, fish heterophils contain large
Morphology of Blood Cells in Nonmammalian Vertebrates
amounts of myeloperoxidase and their macrophages
produce nitric oxide and reactive oxygen.45,53,54
Their morphology is variable, and the nucleus may
range in shape between kidney-like to having two or
three lobes. The cytoplasm contains very thin, pale
eosinophilic granules. Because of the diverse shapes
of fish heterophils, a classification scheme has been
developed that is based on the size and number of
nuclear lobes and shape and staining characteristics
of the granules (Fig 4, d and e).46 The presence of
peroxidase in fish leukocytes has been associated
with bactericidal activity and functions as a defensive
immune mechanism.55
Eosinophils (Fig 4, f) are described for some fish
but are poorly understood. Granules in fish eosino-
phils may have bar-shaped crystalloids similar to
those described in human eosinophils. There is a
lack of knowledge regarding the full function of fish
eosinophils, but they seem to function in a similar
manner to mammalian mucosal mast cells.56 The
eosinophils are found frequently in the digestive
tract and gills and have been associated with anti-
genic stimulation and parasitic infestations.57,58
Basophils are the most variable cell type in fish
(Fig 4, h). They are apparently absent in certain
species, such as zebra fish (Danio rerio)59 and sea bass
(Dicentrarchus labrax),60 but present in others, like
the sea bream (Sparus aurata)61 and carp (Cyprinus
carpio).62 When present, basophils occur in very low
numbers. Fish that do not have basophils also appear
to lack mast cells, immunoglobulin E, and seroto-
nin.63
Lymphocytes are the most common circulating
leukocyte found in fish. They are generally classified
as small (e.g., the predominant form; Fig 4, i) or
large.45 Plasma cells are occasionally found in the
blood of fishes.
T-, B- and NK-lymphocytes are present in bony
and cartilaginous fishes, but not in jawless fishes or
invertebrates.64 It is generally believed that jawless
fishes only have innate immunity.65 However, lym-
phocytes of lampreys have been shown to transcribe
novel variable lymphocyte receptors, consisting of
leucine-rich repeats, that can generate diversity
through a somatic rearrangement process.66 In con-
trast to higher vertebrates, most fish hatch at the
embryonic stage of life, with the innate immune
system serving as their primary defense mechanism.
The acquired immune system (B- and T-cells) arises
later in life, generally after food consumption.67
Monocytes are large leukocytes with an abundant
blue-gray cytoplasm that lacks granules and are oc-
casionally vacuolated (Fig 4, g).45 As in mammals,
monocytes migrate to the tissues and became mac-
95
rophages.68 Aggregates of melanin containing mac-
rophages normally occur within the liver, spleen,
and kidney of fish and are also found in the liver of
reptiles and amphibians.69 In fish, melanomacroph-
age centers have been shown to trap and retain
antigens and immune complexes, thus functioning
as a primitive analog to the lymphoid germinal cen-
ter.70
Thrombocytes
Fish thrombocytes are variable in shape and may be
rounded to oval or spindle shaped. Activation is
frequent in blood smears obtained from the caudal
vein, and some of the changes noted in the cells on
a smear may be caused by the activation of the
clotting cascade (Fig 4, b and c). The nucleus is
usually oval, but may be segmented.28 Ultrastructur-
ally, the fish thrombocyte is similar to that found in
birds and reptiles. As with other vertebrates, fish
thrombocytes are involved in blood clotting, phagocy-
tosis, and other possible immunologic functions.71,72
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3. Baumann R, Dragon S: Erythropoiesis and red cell
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