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Life Sciences
Occasional Papers

Royal Ontario Museum
December 15, 1969

No. 14

ROYAL ONTARIO MUSEUM LIBRARIES

Hemoglobin Electrophoresis in the Systematics
of Bats (Microchiroptera)

by J. R. Tamsitt1 and Dario Valdivieso2

Electrophoresis is a most useful procedure
for systematically comparing proteins of dif-
ferent organisms, and data from electro-
phoretic studies of blood proteins have aided
in the clarification of phylogenetic relation-
ships and in grouping higher taxonomic cate-
gories (Foreman, 1960; Dessauer, 1966;
Johnson, 1968). Hemoglobins, unlike some
serum proteins, are not affected by diet, age,
reproductive state, temperature or other
variables. There are, however, few electro-
phoretic data on bat hemoglobin. Manwell
and Kerst (1966) and Mitchell (1966)
found considerable individual variation and
only minor differences between species in the
hemoglobins of temperate bats of the family
Vespertilionidae. Valdivieso et al. (1969)
compared electrophoretic properties of
hemoglobin from Puerto Rican bats of the
families Phyllostomatidae, Vespertilionidae
and Molossidae and found differences and
similarities correlated with taxonomic af-
finity. Although hemoglobins of closely
related species were indistinguishable, Valdi-
vieso et al. (op. cit.) concluded that hemo-
globin electrophoresis may be used to add
additional evidence to estimates of relation-
ships derived from traditional criteria.

Two distinct electrophoretic phenotypes
of hemoglobin occur in bats of the family
Vespertilionidae (Manwell and Kerst, op.
cit.; Mitchell, op. cit.; Valdivieso et al., op.

1. Department of Mammalogy, ROM.

2. Seneca College, Willowdale, Ontario; Research
Associate, Department of Mammalogy, ROM.

3 1761 05162603 4

cit.), but only one phenotype is known in
bats of the families Phyllostomatidae and
Molossidae (Valdivieso et al., op. cit.). Ex-
tensive sampling of additional taxa should
indicate whether Neotropical phyllostomatid
and molossid bats possess the considerable
hemoglobin polymorphism found in tem-
perate vespertilionid bats, whether there are
geographic differences in hemoglobins of
disjunct populations of the same taxa, and
should help determine the worth of hemo-
globin electrophoretic properties in system-
atic studies of microchiropteran bats. We
therefore describe here results of new elec-
trophoretic studies on bat hemoglobin which
confirm and extend previous observations.

Materials and Methods — Hemoglobin sam-
ples were obtained from 109 bats of 13
species collected in Colombia, Venezuela
and Ontario in November and December,
1968, and January, 1969 (Table I). Bats
were taken in "mist" nets or from culverts
in Colombia and Venezuela and from a mine
in Ontario. Identifications were made by
R. L. Peterson and the senior author (JRT),
and specimens are deposited in the mammal
collection of the Royal Ontario Museum.
The large Artibeus which occurs sympatri-
cally with A. lituratus in west central
Colombia was referred by Tamsitt and
Valdivieso (1963: 175) to A. jamaicensis
jamaicensis, but specimens (ROM 48959,
48981-48986) are indistinguishable from
examples from Trinidad described by

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ROYAL ONTARIO MUSEUM

Andersen (1906: 420) as A. planirostris
trinitatis. Although Hershkovitz (1949) con-
sidered A. planirostris to be synonymous
with A. jamaicensis, the allocation of this
large fruit bat to the species A. jamaicensis
is questionable (R. L. Peterson, personal
communication). Until the taxonomic status
of the large Artibeus from northern South
America is clarified, we therefore only tenta-
tively refer our specimens from the Magda-
lena River Valley of Colombia to A . j. trini-
tatis.

Blood was obtained by heart puncture
with sterile syringes rinsed with heparin,
transferred to 0.5 ml tubes, and centrifuged
at 3,000 rpm for 20 minutes to separate
plasma from the packed cells. The buffy
coat was removed by aspiration with a
micropipette. The cells were washed once
with cold 0.85 per cent saline and then
centrifuged. The hemolysate was prepared
by adding 1.5 volumes of distilled water to
the packed cells. After thorough mixing, the
solution was frozen and thawed three times.
The hemolysates were then centrifuged at
4,000 rpm for five minutes, and the super-
natant was stored at — 5°C. Samples were
run as oxyhemoglobin. Before electrophore-
sis the samples were thawed, shaken with
half volume of chloroform, and centrifuged
in a Beckman Microfuge for two minutes at
14,000-17,000 rpm to extract methemo-
globin and to improve electrophoretic resolu-
tion. Electrophoretic mobility characteristics
of hemoglobins did not change during cold
storage, and patterns were reproducible after
two months. Although stabilizing procedures,
e.g., derivitization of oxyhemoglobins to
carbomonoxy- or cyanomet-forms, have been
employed for hemoglobins, these methods
were not conveniently adaptable to field con-
ditions and were the motive for our decision
to base electropherograms on mobility
characteristics of oxyhemoglobins.

Hemolysates were analyzed by electro-
phoresis in the Model R-101 Microzone Cell
(Beckman Instrument Co., Fullerton, Cali-
fornia) with Sepraphore III poly acetate
membranes (Gelman Instrument Co., Ann
Arbor, Michigan) and barbital buffer pH
8.6, ionic strength 0.075. The sample (0.25
jA) was applied to the membrane with a
Beckman applicator for 12 seconds, and

separation was accomplished at 200 V for
20 minutes. Human hemoglobins A, C and S
were run simultaneously with bat hemo-
globin as controls. After electrophoresis,
hemoglobins were stained with Ponceau S.

Results — Only one electropherogram pat-
tern, a single anodal band, was seen in the
13 species at pH 8.6 (Figs. 1-3). The
homogeneous hemoglobin components of
Phyllostomus discolor, P. hastatus, Glosso-
phaga soricina, Lonchophylla robusta,
Carollia perspicillata, Sturnira lilium, Arti-
beus phaeotis, A. cf. jamaicensis, A. lituratus
and Desmodus rotundus were identical and
indistinguishable from human S. Hemo-
globins of Myotis lucijugus and M. subulatus
(Fig. 3, A and C) were identical and had
mobilities corresponding to human C. The
hemoglobin of Molossus molossus (Figs. 2,
E, and 3, C) migrated the least and had a
mobility slightly more cathodal than human
C. Neither clear genetic polymorphism nor
intrapopulation or interspecific variation was
found in any of the taxa, but only a few
individuals of some species were tested, and
further sampling may reveal variant patterns.
Moreover, hemoglobins of G. soricina (Fig.
2, D), C. perspicillata (Fig. 1, C) and A.
lituratus (Fig. 1, D and E), collected in Co-
lombia and Venezuela from localities sepa-
rated by the Eastern Andean Cordillera and
approximately 1,000 airline kilometers
apart, were indistinguishable, as were those
of P. discolor and A. lituratus from Co-
lombian localities 55 and 85 airline kilo-
meters distant in the Magdalena River Val-
ley and the western slopes of the East
Andean Cordillera. No relationship was evi-
dent between pattern types and sex, age
(neonate, juvenile, young adult or adult) or
in female reproductive condition (non-
parous, pregnant, lactating or post-lactat-
ing). Although fetal hemoglobin distinct
from adult hemoglobin occurs in some mam-
mals (reviewed by Manwell, 1960), patterns
for juvenile and adult D. rotundus (Fig. 2,
A) and for fetal and adult P. discolor (Fig.
2, B) do not differ.*

*The reduced amount and lower concentration of
fetal hemoglobin account for the differences seen in
Fig. 2, B in the width of the hemoglobin bands of
adult and fetal P. discolor.

Discussion — We encountered three electro-
phoretically different hemoglobins in this
study. With the exception of the vampire bat
(D. rotundus), there was in general a cor-
relation of hemoglobin pattern with taxono-
mic relationship. One electrophoretic pat-
tern, an anodal band corresponding to
human S, was found in genera and species
of the subfamilies Phyllostomatinae, Glosso-
phaginae, Carolliinae, Sturnirinae and Steno-
derminae of the family Phyllostomatidae
(Table II, Fig. 4). This hemoglobin electro-
phoretic pattern is identical to that reported
by Valdivieso et al. (1969) for the phyllo-
stomatid genera Monophyllus (subfamily
Glossophaginae), Artibeus, Stenoderma
(subfamily Stenoderminae) and Erophylla
(subfamily Phyllonycterinae) from Puerto
Rico. Excluding Chilonycteris (subfamily
Chilonycterinae), whose hemoglobin is dif-
ferent (Valdivieso et al., op. cit.) and whose
status as a subfamily of the Phyllo-
stomatidae has been questioned (Machado-
Allison, 1967), the hemoglobins of the 13
species of phyllostomatids analyzed to date
are invariably the same (Table II). We
therefore find a reasonable amount of sup-
port from "classical" sources (Miller, 1907)
for a close relationship among the sub-
families of the phyllostomatid complex as
defined by hemoglobin electropherograms.

The relationship of one genus, Sturnira,
to other Phyllostomatidae is still not clear.
Although Miller (op. cit.) placed the genus
in a separate subfamily because of its highly
specialized tooth structure, de la Torre
(1961) contended that Sturnira should be
placed with the Stenoderminae. On the basis
of chromosome similarities, Baker (1967)
and Gardner and O'Neill (1969) concurred
with de la Torre, as did Wenzel et al.
(1966) after a study of parasitic flies (Streb-
lidae) and their hosts. The morphology of
sturnirine spermatozoa, however, differs
from that of other phyllostomatids (Forman,
1968), and certain mites (Spinturnicidae)
parasitizing Sturnira are unique to the group
(Machado- Allison, 1965). Although our
limited hemoglobin data do not distinguish
Sturnira from other phyllostomatids, the
relationship of the sturnirines with the
carollines (see Walton and Walton, 1968),
on the one hand, and with the glossophagines

(see Baker, 1967), on the other, needs clari-
fication before subfamilial status is disre-
garded and the group is allied with the steno-
dermines.

An interesting finding was that the hemo-
globin of the vampire bat (Desmodus
rotundus) was electrophoretically indistin-
guishable from those of bats of the family
Phyllostomatidae. Although extensive modi-
fications, reflecting adaptations to sangui-
vorous feeding habits, indicate considerable
temporal isolation of the desmodontids from
the phyllostomatids, evidence is accumulat-
ing to suggest that vampire bats are phy-
letically more closely allied to members of
the family Phyllostomatidae than is implied
by current classification. Miller (1907), and
subsequently Simpson (1945), Cabrera
(1958), Hall and Kelson (1959) and
Anderson and Jones (1967), recognized the
Desmodontidae, but Dobson (1875), Winge
(1941-42) and Bourliere (1955) con-
sidered the desmodontids as a subfamily of
the Phyllostomatidae (cf. Walton and
Walton, 1968, for a recent summary of early
nomenclatorial history and synonomy). Data
from host-ectoparasite relationships (Ma-
chado-Allison, 1967; Wenzel et al, 1966),
spermatozoa morphology (Forman et al.,
1968), and karyotypes (Hsu and Benirschke,
1967; Forman et al., op. cit.) indicate a
close relationship between the desmodontids
and the Phyllostomatidae. Moreover, the
degree of development of the neocortex of
Desmodus is comparable to phyllostomatids
(Mann, 1963), as are orientation sounds
and behaviour (Griffin and Novick, 1955;
Novick, 1963), palate and wing structure
(Miller, op. cit.) and post-cranial skeletal
elements (Walton and Walton, op. cit.). Al-
though the hemoglobin morphs of Desmodus
and nine genera of six subfamilies of the
Phyllostomatidae are identical (Table II;
Fig. 4), peptide mapping might show that
these specific hemoglobins differ in primary
structure, as Foreman (1964) has shown in
rodents with nearly identical hemoglobin
ionograms. In our opinion a detailed study
of hemoglobin and additional biochemical
characters of the involved genera (Des-
modus, Diaemus, Diphylla) is needed before
vampire bats are assigned as a subfamily of
the family Phyllostomatidae.

A second electrophoretic pattern was seen
in bats of the family Vespertilionidae. In
Myotis lucifugus and M. subulatus from
Ontario, hemoglobin migration was the same
and approximately equal to human C (Table
II, Fig. 4). A comparable hemoglobin was
found by Mitchell (1966) in M. lucifugus
and M. grisescens from Missouri. Manwell
and Kerst (1966), on the other hand, found
a slowly moving major and a more rapidly
moving anodal zone in both M. lucifugus and
M. keenii from Illinois. Populations of M.
lucifugus from Missouri, Illinois and Ontario
represent the same subspecies (M. /. luci-
fugus), and in the absence of intra-
populational variation, these geographically
different phenotypes are not easily interpreted
unless hemoglobin is polymorphic in this
species. Comparable geographic differences
have been found in Eptesicus fuscus. Hetero-
geneous hemoglobins were found by Manwell
and Kerst (1966) in E. f. fuscus from Illi-
nois and by Valdivieso et al. ( 1969) in E. f.
wetmorei from Puerto Rico, but Mitchell
(op. cit.) found a homogeneous hemoglobin
in Missouri E. f. fuscus. In other vesper-
tilionids, e.g., Pipistrellus subflavus and
Plecotus townsendii, however, similar het-
erogeneous hemoglobins have been found
in both Illinois and Missouri. Differences in
electrophoretic technique could account for
these differences, but this is unlikely as
Mitchell (op. cit.) and Valdivieso et al.
(op. cit.) used the same electrophoretic pro-
cedure (cellulose polyacetate) but obtained
different results. A more logical explanation
to account for the presence of more than
one adult hemoglobin in M. lucifugus and
E. fuscus is polymorphism. But the numbers
of Eptesicus and Myotis and the geographic
areas sampled to date have been small, and
extensive sampling throughout the range of
these and other vespertilionids is needed
to determine the degree of allelic variation
and also to determine whether we are cor-
rect in attributing these differences to poly-
morphism.

A third electrophoretic pattern, a slow
anodal band, was seen in Molossus molossus
of the family Molossidae (Table II, Fig. 4).
This same pattern has been found as well
in the molossid bats Tadarida brasiliensis
(Johnson and Wicks, 1959) and in M. m.

fort is from Puerto Rico (Valdivieso et al.,
1969). Although serum proteins (Johnson
and Wicks, op. cit.) and immunologic data
(Forman et al., 1968) do not differen-
tiate the Molossidae from the Vespertilioni-
dae, hemoglobin and karyotype differences
(Baker and Patton, 1967; Wainberg, 1966)
support Miller's (1907) interpretation that
the two groups of insectivorous bats are
sufficiently distinct to merit familial separa-
tion. Furthermore, lactate dehydrogenase
patterns of Molossus and Eptesicus tissues,
although similar, are distinguishable and
different from those of phylostomatid bats
(unpublished data).

The similarities and differences we have
found among bat hemoglobins correspond
in large part to the phylogeny presented by
Simpson (1945). The hemoglobins of mem-
bers of the superfamilies Phyllostomatoidea
and Vespertilionoidea differ from each other
more than the hemoglobins of bats within
each of these groups differ. Except for the
Chilonycterinae, evidence to date suggests
that the hemoglobins of the Phyllostoma-
toidea are quite similar. Hemoglobins of the
Vespertilionoidea, on the other hand, appear
to differ among themselves far more than do
the hemoglobins of the Phyllostomatoidea.

Recent publications (Thompson et al.,
1966; Foreman, 1968; Rasmussen et al.,
1968; Self A., 1968) have revealed not only
hemoglobin variation between species of the
same genus but also the occurrence of poly-
morphism within a given species in the order
Rodentia. This type of variation, however,
is not always the case, as there is a remark-
able homogeneity between the hemoglobins
of Microchiroptera (Manwell and Kerst,
1966; Valdivieso et al, 1969; this study).
Because the rate of hemoglobin change is
low, based on the amount of variation found
within and between closely related genera,
similarities in bat hemoglobin electropho-
retic patterns, although of limited taxonomic
value, may be indicative of phylogenetic
relationships. Hemoglobin analysis certainly
supports the validity of the common origin
of the Phyllostomatidae and the Desmodon-
tidae, whose genera have hemoglobins indis-
tinguishable from one another but different
from those of the Vespertilionidae and
Molossidae.

+

D
E

* ♦

3 4

+

6 7 8

Figure 1

Electrophoretic patterns of erythrocyte hemolysates

at pH 8.6. Arrows indicate baselines of sample

application.

A. Carollia perspicillata (Venezuela): 1-3,
5-8, $ $ ; 4, Human A.

B. C. perspicillata (Venezuela): 1-3, 5-8, 2 ?;
4, Human S.

C. C. perspicillata: 1, 2, $ $ (Colombia); 3,

$ (Venezuela); 4, Human S; 5-8, 5 2
(Venezuela).

D. Artibeus lituratus: 1-4, S S (Colombia);
5-7, S S (Venezuela); 8, Human CA.

E. A. lituratus: 1, 2 (Colombia); 2-4, 9 2
(Venezuela); 5-7, $ S (Venezuela); 8,
Human CA.

F. A. cf. jamaicensis: 1-3, 2 2 (Colombia);
4, 5, $ $ (Colombia); A. phaeotis: 6, 7,
S $ (Colombia); 8, Human CA.

Acknowledgments — We are indebted to Drs.
Omar Linares, Francis R. Hunter and
M. Brock Fenton for laboratory and field
assistance; to the staffs of the Depart-
ments of Biology, Universidad de la Region
Centro-Occidental, Barquisimento, Univer-
sidad de los Andes, Bogota, and the In-
stituto de Zoologia Tropical, Universidad
Central, Caracas, for providing us with
laboratory facilities; to Dr. Gonzalo Medina
Padilla, Director, Rancho Grande Biological
Station, for his co-operation and the use of

facilities; and to the staff of the Department
of Mammalogy, Royal Ontario Museum, for
their assistance. The illustrations were pre-
pared by Mrs. Sophie Poray and Mr. L.
R. Warren, Royal Ontario Museum. We
thank Dr. R. L. Peterson for aid in identify-
ing specimens and for critically reading
the manuscript. These activities were sup-
ported by the National Research Council
of Canada, the Canadian National Sports-
men's Show and the Royal Ontario Museum.

+

D

+

+

6

Figure 2

Electrophoretic patterns of erythrocyte hemolysates

at pH 8.6. Arrows indicate baselines of sample

application.

A. Desmodus rotundus (Colombia): 1, juve-
nile 8 ; 2-4, 9 5 ; 5, Human CA.

B. Phyllostomus discolor (Colombia): 1, $
fetus; 2-4, $ $ ; 5, Human CA.

C. P. hastatus (Venezuela): 1, $ ; 2-4, 9?;
5, Human CA.

D. Glossophaga soricina: 1-3, $ $ (Vene-
zuela); 4, ? $ (Venezuela); 5, 9 (Colom-
bia); 6, Human CA.

E. Molossus molossus (Colombia): 1, 2, $ $;
3-5, 9 9 ; 6, Human CA.

Summary — Hemoglobins of 109 examples
of 1 3 species of bats from Colombia, Vene-
zuela and Canada were compared by cellu-
lose polyacetate electrophoresis. All bats
exhibited a single-banded hemoglobin pheno-
type. The phyllostomatid bats Phyllostomus
discolor, P. hastatus (subfamily Phyllosto-
matinae), Glossophaga soricina, Loncho-
phylla robusta (subfamily Glossophaginae),
Carollia perspicillata (subfamily Carolli-
inae), Sturnira lilium (subfamily Sturniri-
nae), Artibeus phaeotis, A. cf. jamaicensis

and A. lituratus (subfamily Stenoderminae)
had indistinguishable hemoglobin morphs.
Hemoglobin electropherograms of the vam-
pire bat, Desmodus rotundus (family Des-
modontidae), and the phyllostomatid bats
could not be differentiated, suggesting a
closer relationship of the desmodontid to the
phyllostomatid bats than is implied by cur-
rent classification. The hemoglobins of the
vespertilionid bats Myotis lucijugus and M.
subulatus were identical but differed from
the phyllostomatid and desmodontid bats by

ir»

+

B

+

2 3 4 5 6

Figure 3

Electrophoretic patterns of erythrocyte hemolysates

at pH 8.6. Arrows indicate the baselines of sample

application.

A. 1, Lonchophylla robusta 2 (Colombia); 2,
Sturnira lilium $ (Colombia); 3, S. lilium

2 (Colombia); 4, Artibeus cf. jamaicensis
$ (Colombia); 5, Myotis subulatus $
(Ontario); 6, Human CA.

B. 1, Phyllostomus hastatus $ (Venezuela);

2, Glossophaga soricina $ (Venezuela); 3,
Carollia perspicillata S (Venezuela); 4,
S. lilium S (Colombia); 5, A. lituratus 2
(Venezuela); 6, Human CA.
1, A. lituratus $ (Venezuela); 2, Desmodus
rotundus 2 (Colombia); 3, Molossus
molossus $ (Colombia); 4, Myotis luci-
fugus S (Ontario); 5. M. molossus 2
(Colombia); 6, Human CA.

a slower migration. A slow, almost isoelec-
tric hemoglobin characterized the molossid
bat Molossus molossus and distinguished it
from bats of the families Phyllostomatidae,
Desmodontidae and Vespertilionidae.

Electropherograms were independent of
sex and age, and there was no variation in
the electrophoretic patterns of individuals
of a species from the same or different
localities. Electrophoretic properties of bat
hemoglobin have limited taxonomic value at
lower levels of organization but are useful in
phylogenetic analysis.

Resumen — Se comparan las hemoglobinas
de 109 ejemplares correspondientes a 13
especies de quiropteros de Colombia, Vene-
zuela y Canada por medio de electroforesis
de poliacetato de celulosa. El fenotipo exhi-
bido por todos los murcielagos es de una
banda simple de hemoglobina. Los quirop-
teros filostomatidos Phyllostomus discolor,
P. hastatus (subfamilia Phyllostomatinae),
Glossophaga soricina, Lonchophylla robusta
(subfamilia Glossophaginae), Carollia pers-
picillata (subfamilia Carolliinae), Sturnira
lilium (subfamilia Sturnirinae), Artibeus

phaeotis, A. cf. jamaicensis and A. litura-
tus (subfamilia Stenoderminae) presentan
morfos identicos de hemoglobinas. Los elec-
troferogramas del vampiro, Desmodus rotun-
dus (familia Desmodontidae), y de los
filostomatidos no pueden diferenciarse, lo
ciial sugiere un parentesco mas cercano
entre los desmodontidos y filostomatidos de
lo implicado en la clasificacion actual. Las
hemoglobinas de los vespertilionidos Myotis
lucijugus y M. subulatus son identicas
pero difieren de las de los filostomatidos y
de los desmodontidos por presentar migra-
ciones mas lentas. Una hemoglobina lenta,
casi isoelectrica, caracteriza al molosido
Molossus molossus distinguiendolo de los
filostomatidos, desmodontidos y vesperti-
lionidos.

Diferencias en sexo y edad no afectan los
patrones de los electroferogramas. No existe
variacion en los modelos electroforeticos de
ejemplares de una especie ya sea de la
misma o de diferentes localidades. Las pro-
piedades electroforeticas de la hemoglobina
quiropteriana tienen valor taxonomico limi-
tado en niveles de organizacion bajos pero
son de utilidad en analisis filogenetico.

HUMAN CA

HUMAN CS

FAMILY PHYLLOSTOMATIDAE

SUBFAMILY PHYLLOSTOMATINAE

Phyllostomus discolor
Phyllostomus hastatus

SUBFAMILY GLOSSOPHAGINAE

Glossophaga soricina
Lonchophylla robusta

SUBFAMILY CAROLLIINAE

Carollia perspici llata

SUBFAMILY STURNIRINAE

Sturnira lilium

SUBFAMILY STENODERMINAE

Artibeus phaeotis
Artibeus cf . jamaicensis

II

II

Artibeus lituratus

FAMILY DESMODONTIDAE
Desmodus rotundus

Figure 4

Intergeneric and interspecific comparison of hemo-
globin morphs of bats. The circle represents the
origin.

FAMILY VESPERTILIONIDAE
Myotis lucifugus

Myotis subulatus

FAMILY MOLOSSIDAE
Molossus molossus

I
I

I

TABLE I

Collection data for bat species examined in this study

Age and

Number of

Species

Sex

Specimens

Date

Locality

Phyllostomus discolor

Adult 9

1

December 18, 1968

1

Young Adult cf

1

January 8, 1969

4

Adult cf

1

55

5

Pregnant 9

1

59

5

Fetus cf

1

59

5

Phyllostomus hastatus

Adult cf

3

November 19, 1968

6

Adult 9

3

"

6

Glossophaga soricina

Adult cf

4

November 18, 1968

7

Adult 9

1

"

7

Adult 9

1

December 18, 1968

1

Pregnant 9

1

January 8, 1969

4

Lonchophylla robusta

Adult 9

1

95

4

Carollia perspicillata

Adult cf

7

November 18, 1968

7

Young Adult cf

1

"

7

Adult 9

11

55

7

Young Adult 9

3

55

7

Adult cf

1

December 18, 1968

1

Young Adult cf

1

December 28, 1968

2

Adult cf

4

January 8, 1969

5

Pregnant 9

4

55

5

Adult 9

6

59

5

Sturnira lilium

Adult cf

1

December 18, 1968

1

Adult 9

1

"

1

Artibeus phaeotis

Adult cf

1

January 8, 1969

4

Artibeus cf. jamaicensis

Adult cf

3

55

4

Pregnant 9

1

95

4

Lactating 9

2

95

4

Artibeus lituratus

Adult cf

8

November 19, 1968

6

Adult 9

5

55

6

Adult cf

7

December 18, 1968

1

Pregnant 9

1

December 22, 1968

3

Adult cf

1

January 8, 1969

5

Desmodus rotundas

Adult cf

3

December 26, 1968

5

Adult 9

2

"

5

Pregnant 9

2

99

5

Adult cf

2

99

5

Neonate cf

1

"

5

Myotis lucifugus

Adult cf

4

December 7, 1968

8

Adult 9

1

"

8

Myotis subulatus

Adult cf

1

"

8

Molossus molossus

Adult cf

2

December 22, 1968

3

Adult 9

2

99

3

Lactating 9

1

"

3

KEY TO LOCALITIES
Colombia

1. Cundinamarca : Mesitas del Colegio (1210 m).

2. " : Pacho (1859 m).

3. " : Villeta (804 m).

4. Tolima: Melgar (430 m).

5. : 9 km NW Melgar (approx. 430 m).
Venezuela

6. Aragua: Portachuelo Pass, Rancho Grande (1100 m).

7. Lara: 7 km E Barquisimeto (566 m).
Canada

8. Ontario : Hunt Mine, Renfrew Co. (approx. 90 m).

TABLE II

Grouping of species of bats by hemoglobin electropherograms.* Heterogeneous hemoglobins
are designated by the subscript "1" for the anodal band and "2" for the cathodal band

Origin

Zone

Pipistrellus subflavus^
Eptesicus fuscus^
Tadarida brasiliensis
Molossus molossus

Human Hb-C

Myotis lucifugus
My otis grisescens
Myotis subulatus
Pipistrellus subflavus\

Chilonycteris
parnellii

Human Hb-S

Phyllostomus discolor
Phyllostomus hastatus
Glossophaga soricina
Lonchophylla robusta
Monophyllus redmani
Carol! ia perspicillata
Sturnira lilium
Artibeus phaetois
A rtibeus jamaicensis
Artibeus cf. jamaicensis
Artibeus lituratus
Stenoderma rufum
Erophylla bombifrons
Desmodus rotundus
Eptesicusfuscusi

Human Hb-A

*Data from Johnson and Wicks (1959), Mitchell (1966), Valdivieso et al. (1969) and this study. Published
information from species whose hemoglobins were subjected to electrophoresis with reference hemoglobins
other than human are not included in the table.

Literature Cited

Andersen, Knud

1906 Brief diagnoses of a new genus and ten new forms of stenodermatous bats. Ann.
Mag. Nat. Hist., vol. 18, ser. 7, pp. 419-423.
Anderson, Sydney, and J. Knox Jones, Jr., eds.

1967 Recent mammals of the world; a synopsis of families. Ronald Press Co., New
York, viii + 453 pp.
Baker, Robert J.

1967 Karyotypes of bats of the Family Phyllostomidae and their taxonomic implica-
tions. S. West. Nat., vol. 12, no. 4, pp. 407-428.
Baker, Robert J., and James L. Patton

1967 Karytotypes and karyotypic variation of North American vespertilionid bats. /.
Mammal., vol. 48, no. 2, pp. 270-286.
Bourliere, F.

1955 Systematique. In Grasse, Pierre-P., ed. Traite de Zoologie. Paris, Masson et Cie.
tome 17, pp. 1806-1844.
Cabrera, Angel

1958 Catalogo de los mamiferos de America del Sur. 1. ( Metatheria — Unguiculata —
Carnivora). Rev. Mus. Argent. Cienc. Nat. "Bernardino Rivadavia". Zool. vol.
4, 1957 (1958), pp. 1-307.
Dessauer, Herbert C.

1966 Taxonomic significance of electrophoretic patterns of animal sera. Bull. Serol.
Mus., New Brunswick, no. 34, pp. 4-8.

10

Dobson, G. E.

1875 Conspectus of the suborders, families, and genera of Chiroptera arranged accord-
ing to their natural affinities. Ann. N.H., ser. 4, no. 16, pp. 345-357.
Foreman, Charles W.

1960 Electromigration properties of mammalian hemoglobins as taxonomic criteria.
Amer. Midi. Nat., vol. 64, no. 1, pp. 177-186.

1964 Tryptic peptide patterns of some mammalian hemoglobins. /. Cell. Comp. Physiol.,
vol. 63, no. 1, pp. 1-6.

1968 Hemoglobin ionographic properties of Peromyscus and other mammals. Comp.
Biochem. Physiol, vol. 25, no. 2, pp. 727-731.
Forman, G. Lawrence

1968 Comparative gross morphology of spermatozoa of two families of North American
bats. Kans. Univ. Sci. Bull., vol. 47, no. 16, pp. 901-928.
Forman, G. Lawrence, Robert J. Baker and Jay D. Gerber

1968 Comments on the systematic status of vampire bats (Family Desmodontidae).
Syst. Zool., vol. 17, no. 4, pp. 417-425.

Gardner, Alfred L., and John P. O'Neill

1969 The taxonomic status of Sturnira bidens (Chiroptera: Phyllostomidae) with notes
on its karyotype and life history. Louisiana State Univ., Mus. Zool., Occ. Pap. no.
38, pp. 1-8.

Griffin, Donald R., and Alvin Novick

1955 Acoustic orientation of Neotropical bats. /. Exp. Zool., vol. 130, no. 2, pp. 251—
299.
Hall, Eugene Raymond, and Keith R. Kelson

1959 The mammals of North America. Ronald Press Co., New York. vol. 1. xxx + 546
+ 10 + 79 pp.
Hershkovitz, Philip

1949 Mammals of northern Colombia. Preliminary report no. 5: Bats (Chiroptera).
Proc. U.S. Natn. Mus., vol. 99, no. 3246, pp. 429-454.
Hsu, T. C, and K. Benirschke, eds.

1967 An atlas of mammalian chromosomes, vol. 1. Springer- Verlag, New York. 50
folios.

Johnson, Murray L.

1968 Application of blood protein electrophoretic studies to problems in Mammalian
taxonomy. Syst. Zool., vol. 17, no. 1, pp. 23-30.

Johnson, Murray L., and Merril J. Wicks

1959 Serum protein electrophoresis in mammals - taxonomic implications. Syst. Zool,
vol. 8, no. 2, pp. 88-95.

Machado-Allison, C. E.

1965 Las especies venezolanas del genero Periglischrus Kolenati 1857 (Acarina, Mesos-
tigmata, Spinturnicidae). Acta Biol. Venez., vol. 4, art. 11, pp. 259-348.

1967 The systematic position of the bats Desmodus and Chilonycteris, based on host-
parasite relationships (Mammalia; Chiroptera). Proc. Biol. Soc. Wash., vol. 80,
pp. 223-226.
Mann, Guillermo

1963 Phylogeny and cortical evolution in Chiroptera. Evolution, Lawrence, Kansas, vol.
17, no. 4, pp. 589-591.
Manwell, Clyde

1960 Comparative physiology: blood pigments. A. Rev. Physiol, vol. 22, pp. 191-244.
Manwell, Clyde, and K. V. Kerst

1966 Possibilities of biochemical taxonomy of bats using hemoglobins, lactate dehydro-
genase, esterases and other proteins. Comp. Biochem. Physiol, vol. 17, no. 3, pp.
741-754.

11

Miller, Gerrit S., Jr.

1907 The families and genera of bats. U.S. Natn. Mus. Bull. 57, pp. 1-282.
Mitchell, Henry A.

1966 Multiple hemoglobins in bats. Nature, London, vol. 210, no. 5040, pp. 1067-1068.
Novick, Alvin

1963 Orientation in Neotropical bats. II. Phyllostomatidae and Desmodontidae. /. Mam-
mal., vol. 44, no. 1, pp. 44-56.
Rasmussen, David I., J. Neil Jensen and Richard K. Koehn

1968 Hemoglobin polymorphism in the deer mouse, Peromyscus maniculatus. Biochem.
Genetics, vol. 2, no. 1, pp. 87-92.
Self Ahl, Alwynelle

1968 Electrophoretic examination of hemoglobin and plasma proteins from three alti-
tude groups of Peromyscus maniculatus nebrascensis. Com p. Biochem. Physiol., vol.
24, no. 2, pp. 427-435.

Simpson, George Gaylord

1945 The principles of classification and a classification of mammals. Amer. Mus. Nat.
Hist. Bull. 85, pp. 1-350.
Tamsitt, J. R., and Dario Valdivieso

1963 Records and observations on Colombian bats. /. Mammal., vol. 44, no. 2, pp.
168-180.
Thompson, Robert B., H. B. Hewett, S. S. Kilgore, A. P. Shepherd and W. N. Bell
1 966 Haemoglobin variants in a species of wild mice — Peromyscus maniculatus. Nature,
London, vol. 210, no. 5040, pp. 1063-1064.
Torre, Luis de la

1961 The evolution, variation, and systematics of the Neotropical bats of the genus
Sturnira. Ph.D. Thesis, Univ. Illinois. 146 pp.
Valdivieso, Dario, J. R. Tamsitt and Encarnita Conde-del Pino

1969 Electrophoretic properties of Neotropical bat hemoglobin. Comp. Biochem. Physiol.,
vol. 30, no. 1, pp. 117-122.

Wainberg, Ricardo Luis

1966 Cytotaxonomy of South-American Chiroptera. Arch. Biol., vol. 77, no. 3, pp.
411-423.
Walton, Dan W., and Gloria M. Walton

1968 Comparative osteology of the pelvic and pectoral girdles of the Phyllostomatidae
(Chiroptera; Mammalia). /. Grad. Res. Center, Dallas, vol. 37, no. 1, pp. 1-35.
Wenzel, Rupert L., Vernon J. Tipton and A. Kiewlicz

1966 The streblid batflies of Panama (Diptera Calypterae: Streblidae). In Ectoparasites
of Panama, Rupert L. Wenzel and Vernon J. Tipton, eds. Field Mus. Nat. Hist.,
Chicago, pp. 405-675.
Winge, Herluf

1941-42 The interrelationships of the mammalian genera, vol. 1. Monotremata, Mar-
supialia, Insectivora, Chiroptera, Edentata. Translated from the Danish by E. Deich-
mann and G. M. Allen. Edited by Ad. S. Jensen, R. Sparck and H. Volsve,
managing editor. C. A. Reitzel Forlag, K0benhavn. 418 pp.

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