ERYTHROCYTE

MEASUREMENTS
IN FISHES,
AMPHIBIA,
AND REPTILES'
F. A. HARTMAN AND M. A. LESSLER
Department of Physiology, The Ohio State University, Columbus 10, Ohio

Gulliver (1875) made an extensive study of the size of the red cells in verte
brates, extending over a period of years. Cullen ( 1903 ) gives values for the skate
and dogfish, while Cleland and Johnston ( 1912) reported a considerable series,
especially birds. Although Wintrobe ( 1961) tabulated the results of many investi
gators on red cell size, there have been relatively few reports of new red cell
measurements

and many

species

remain

unreported.

The size and shape of red cells gives an indication of the surface available for
the exchange of gases in respiratory functions.
Study of erythrocytes in many dif
ferent species provides an interesting comparison of red cell size in relation to
activities and habits.
While collecting specimens for adrenal studies, we were
frequently

able to n-make fresh blood smears

and later stained

United States.

and studied.

from the animals.

TI-mematerial

was obtained

These

were air-dried

in Panama

and ti-me

MATERIALS AND METHODS

Blood smears were made immediately after the death of the animal. These were
air-dried promptly and stored until prepared for study in the laboratory.
Staining
was carried out under controlled conditions.
Five drops of standard Wright's stain
were allowed to remain on the slide for one minute before addition of a pH 6.5
buffer.

The slide was allowed

to stand

for five minutes

at room

temperature

and

then washed with distilled water for 30 seconds and allowed to dry.
Ten cells on each slide, selected

for excellence

of staining

and internal

cytology,

were measured by means of a calibrated eyepiece used in conjunction with an oil

immersionobjective.In thisway ten measurementsof maximum cellwidth and
length, as well as nuclear width and length, were made. The ratios of cytosome
length to width and nucleus length to width have been calculated.
This is a
measure of cell and nuclear deviation from the spherical shape.
Erythrocytes
of eight species among seven families of fishes, five species among
five families of Amphibia,
and thirty-three
species among seven families of reptiles
were n-measured (Table I).
When measurements
were made of only one individual
of a species, the standard
error of values for ti-medifferent cells is shown.
When
Fisheswere collected at the Marine Biological Laboratory, Woods Hole, Massachusetts.
Other material was obtained in Ohio; in Florida with the Archbold Biological Station as a base;
and in Panama under the auspices of the Smithsonian Institution. We are indebted to Richard
Archbold for his assistance, to Dr. Carl Johnson for the use of the Juan Mina Field Station of
the Gorgas Memorial Institute and to Alois Hartmann for facilities near Volcan in West
Panama.
83

84

F. A. HARTMAN AND M. A. LESSLER

TABLE

Erythrocyte measurements in fishes

L/@NucleusRatio

Family and speciesCytosomeRatio

L/WLength
iiwidth

pLength

@RajidaeRajaerinacea20.3

.31.37DasyatidaeDasyatis

g@Width

1.512.911.578.4

.86.1

.3813.80.241.428.10.246.90.241.17AnguilhidaeAnguila
centrura (4)19.7

E0.37.60.31.513.80.23.2E0.51.19Leptocephalus
rostrata11.5
+1.57.711.434.00.53.30.51.21LabridaeTautogo
conger10.9
0.357.30.181.444.10.153.00.151.37Trigli4aePrionotus
onitis (3)10.5
(4)11.150.157.30.321.534.8+0.283.0d0.l51.37BatrachoididaeOpsanustau14.9
carolinus
212.921.155.8+0.85.211.11Lophii4aeLophius

three or more individual

is l)ased on the averages

+0.5
1.195.1 3.3+0.84.60.5
4.711.11

1.6 2
11.211.13
13.6+1.310.3E1

piscatorius1

specimens of a species were measured,

of the individuals

the standard

1.13

error

involved.

RESULTS
Among

the fishes

studied,

the elasmobranch

erythrocytes

are nearly

twice

the

size of the erythrocytes in the teleosts, and their width is a little more than half their
length. The cytosomes of Opsanus and Lophius are almost as wide as long (ratio
1.1 5 to i .19) . In all of the fishes the ratio of length to width of the nucleus
TABLE

II

Erythrocyte measurements in Amphi&ia

L/WLength
Family and speciesCytosomeRatio
@iAmphiumid,ae
pWidth

L/@NucleusRatio
@Length
pWidth

Amphiumatridactylum

Proteidae
Neeturus maculosus

56.40.77 38.1 0.76

1.48

18.51.04 12.50.30
19.20.91 13.4+0.53

1.48
1.43

30.20.85 17.40.32

1.74

Bufonidae

Bufomarinus

7.40.56
7.60.38

5.20.46
5.60.41

1.42
1.36

Hylidae
20.30.22 13.40.27
1.52
7.40.17 4.50.15
1.64
Hylagratiosa
Ranidae
15.40.401.78 1.8126.40.76
27.90.4536.60.39
10.30.3116.80.45
Rams catesbiana65.31.27
5.60.271.57 1.84

VERTEBRATE

shows

little

range

RED CELL MEASUREMENTS

( 1.11 to 1.37),

the width

being

85

somewhat

less than

the

length.

The tailed Amphibia

study

possess by far the largest

(56 /h to 65 @L
by 37 @&
to.38

this size or smaller ( 19

to width

to 28 .@by 13

in the cytosomes

erythrocytes

@)while the erythrocytes

to 15

is 1.5 or larger.

observed

of the anurans

in our

is one-half

(Table II ) . The ratio of length

The

ratio

of length

to width

in the

nuclei is of similar range.

Among

the reptiles,

in the one species

of crocodilids

studied,

the cytosome

and

nuclear measurements are similar to those in elasmobranchs except that the ratios
of length to width in the cytosomes and nuclei are greater (Table III).
In the
lizards there are considerable differences.
Reptilian red cell lengths range from
TABLE III

Erythrocyte measurements in a crocodilian and in lizards

L/@Length
Family and speciesCytosomeNucleusRatio
@CrocodiidaeCayman
)IWidth

L/WRatio
@Length@iwidth

(3)20.60.5910.90.461.887.10.194.10.571.74IguanidaeAnolis
scierops
carolinensis14.80.4210.30.391.446.00.063.80.181.59Anolis
0.871.847.60.294.80.271.58Anolis
polylepis (3)18.40.4710.1
pachypus15.60.838.30.441.886.10.753.00.172.03Anolis
(3)18.50.3712.00.641.548.00.185.20.101.54Sceloporus
biporcatus
(6)16.90.449.80.271.726.60.253.50.231.89Basiiscus
malachiticus

americanus19.31.04
12.10.471.60 1.547.60.42
6.50.156.40.40
18.60.3111.60.57
5.50.121.19
1.18Iguana
iguana15.40.287.50.372.065.60.244.50.351.34TeiidaeAnadiaocellata16.31.189.00.761.815.70.353.20.271.78Cnemidophoruss

(3)15.00.168.6+0.461.75.5.90.233.5E0.201.66ScincidaeEumecesfasciatus15.90.168.50.161.875.10.163.10.101.67
praesignis

15

to 19

with widths from 7.5

to 12 @.t.Their length to width ratios vary from

i.44 to 2.06. Likewise there is a great range between nuclear measurements

to 8 @)and their length to width ratios are from 1.19 to 2.11.

In general,
larger

from

than

erythrocytes

those

in

of snakes are large ( 15.5

elasmobranchs.

i.2 to 1.86 and the nuclei

The

length

cytosome

to width

ments in three species of Bothrops indicated

of all reptiles studied (Table IV).
In order

to permit

free

movement

ratios

.t

to 23 @&)
; some are even

length

to

from

width

ratios

1.35 to 2.5.

that their erythrocytes

of erythrocytes

(5.1

we can

range

Measure

are the largest

assume

that

the

diameter of the smallest capillaries must be no less than the smaller diameter of
the erythrocyte.
With this assumption it is interesting to compare the minimal
capillary diameters of different species and groups of vertebrates.
The capillaries

86

@
@

F. A. HARTMAN

AND

M. A. LESSLER

of fishes range from 7.3 @a

to 13.8 in diameter.
The largest capillaries are found
in the elasmobranchs
and the smallest among the bony fishes. The capillaries
of Amphibia would range from 12.5 to 13.4 @,
while those of reptiles would range
from 7.5 p@to 13.8 .@. The smallest are found in the I,izards.
DISCUSSION
Cytosotne

size

Since the erythrocyte is the most important carrier of oxygen and carbon
dioxide, its surface area to size ratio is a determining factor in the exchange of these

TABLE

IV

@Erythrocytemeasuremenls in snakes and turtles

Family and speciesCytosomeNucleus@%@Length

pwidth

pColubridaeSpilotes

,@Length

pwidth

pullatus17.70.2410.30.181.727.10.193.70.201.92Dryadophis
boddaerti16.30.5511.00.921.487.00.634.50.831.56Masticophisfiagellum
(3)16.50.2312.60.491.316.80.124.20.111.62Clelia
(3)20.20.4511.00.221.827.30.403.70.111.97Erythrolamprus
delia
bizonus19.80.3413.00.271.528.00.273.70.092.16Leimadophis
epinephalus16.90.9411.70.521.447.70.453.60.332.14Leimadophis

taeniurus19.20.4612.30.131.567.80.343.10.132.52Thalerophis
occidenkilis15.50.2011.70.281.326.00.594.20.321.43Dendrophidion

paucicarinatus17.10.0412.50.761.366.60.703.80.501.73Thamnophis
sauritus16.40.2313.40.271.227.40.174.50.151.64Natrix
sipendon18.20.84
12.30.251.41
1.72Natrix
18.40.3513.81.28
1.496.80.74
7.40.244.50.74
4.30.091.51
taxispilota18.50.2811.70.261.585.80.114.30.121.35Lampropeltis
doliaki18.1
0.844.40.481.61Eiaphe
1.021 1.80.961.537.1
guttaki18.90.2811.80.161.596.20.143.60.131.72Coluber

(4)15.30.1112.10.341.266.70.283.80.081.76Opheodrys
constrictor
aestivus16.30.2912.50.191.306.90.154.60.421.50ViperidaeBoihrops

lateralis22.20.3911.90.171.865.90.124.00.071.47Bothrops
mummifer23.41.0113.60.781.727.50.254.80.501.56Bothrops
atrox22.10.9612.60.461.757.70.554.90.241.57Test
udinidaeGopherus
(3)19.10.6111.20.221.716.70.554.00.121.69Pseudemys
polyphemus

ornala18.60.0111.60.761.605.50.355.20.661.06

gases with the tissues. Thus, a small corpuscle offers the possibility of a greater
rate of exchange than a larger one. Likewise an elliptical body is more efficient than
a spherical one of the same volume. Based strictly on geometrical considerations,
the red cells of the bony fishes would be the most efficient and those of the tailed
amphibians the least efficient. Compared with warm-blooded animals the erythro
cytes of these poikilothermic animals would be much less efficient.

VERTEBRATE

@
@

87

In mammals the non-nucleated disc-shaped red cells have diameters from 5 @&
and 6 ;@(horse, cow, pig, mouse and rat) to about 7.5
(chimpanzee, woodchuck
and llama) (Wintrobe, 1961 ). In birds the nucleated elliptical erythrocytes have
size ranges from 10.7 by 6.1 to 15.8 by 10.2
(Hartman and Lessler, 1963).
The red cell count in poikilotherms is low. Counts of 70,000 to 390,000 per
nim.3 in eiasmobranchs

RED CELL MEASUREMENTS

; 780,000

to 1,490,000

in bony fishes ; 20,000

to 440,000

in

Amphibia ; 560,000 to 1,050,000 in reptiles have been reported.

In homiotherms, the
red cell count in birds ranges between 1,930,000 and 3,690,000 per mm.3 and in
mammals it is much more variable, ranging from 6,300,000 per mm.3 in the
chimpanzee to 18,000,000 per mm.3 in the llama (Wintrobe,
1961).
Cleland and Johnston (1912) measured the erythrocytes in Australian verte
brates. By far the largest erythrocytes they reported were found in the lung fish,
Ceratodus (39 i.&by 25 @&)
. The elasmobranch erythrocyte sizes they reported are
similar to our measurements,
as are the teleost erythrocytes,
except those of
Terapion which are quite small (6
to 7 @a
by 6
to 7 ,@). The erythrocytes of
the lizards Cleland and Johnston measured showed a somewhat greater range in
size than our measurements, but their other reptile erythrocyte measurements are
similar

to those

of our

specimens.

Nuclei

In all fish erythrocytes the nuclei are shorter in proportion to their width than
are the cytosomes.
In the amphibian erythrocyte nuclei there is usually little dif
ference in these ratios. This is also true in one turtle and in Cayman.
In lizards,
six out of ten species possessed relatively rounder nuclei than cytosomes, while in
snakes

all but four

species

had

nuclei

longer

than

the cytosomes.

Nuclei

were

relatively longer than the cytosomes in all but six species of birds, being shorter
in four and doubtful in two (Hartman and Lessler, 1963) . Longer nuclei could
offer a somewhat greater surface for exchange with the cytoplasm.
This may be a
significant factor in nuclear-cytoplasmic
exchange.
Actually the role of the nucleus
in red cell function of the many species with nucleated erythrocytes has never been
determined.
SUMMARY

Measurements of erythrocytes and their nuclei were made in 8 species of fishes.

5 species of Amphibia, and 33 species of reptiles. The cytosomes of elasniobranchs
are almost twice the size of those in teleosts, being 19 to 20 ,u by 12 p. to 14 ,z. The
cytosomes of Opsanus and Lophius are almost as wide as long, while in other fishes
they are much longer than wide. The cytosome measurements in the two anurans,
Hyla and Bufo, are similar to those in elasmobranchs while those in Rana are
larger.
Cytosomes in the crocodile, turtles and some lizards are also similar in
size to those in the elasmobranchs.
The cytosomes of snakes are large (15.5 p. to
23 p.), those of Bothrops being the largest of all reptiles studied.
Based on the
assumption that the least diameter of the erythrocyte indicated the minimal diameter
of ti-mecapillaries, the largest capillaries are found in the elasmobranchs and the
smallest in the teleosts.

88

F. A. HARTMAN AND M. A. LESSLER

LITERATURE

CLELAND,

J.

B.,

AND

T.

H.

JOHNSTON,

1912.

CITED

Relative

dimensions

of

red

blood

cells

of

verte

brates, especially birds. Emu, 11: 188197.

CULLEN, E. K., 1903.

Oval and round

cells in blood of skate and dogfish.

Bull. Johns

Hopkins

Hosp., 14: 352.

GULLIvER, G., 1875.

Observations

on the size and shapes

of ti-mered corpuscles.

Proc.

Zoo!.

Soc. London, 1875: 474495.

HARTMAN,

F.

A.,

AND

M.

A.

LESSLER,

1963.

Erythrocyte

measurements

in

80: 467473.
WINTROBE, M. M., 1961.

Clinical

Hematology.

Lea and Febiger,

Philadelphia.

birds.

The

Auk,