Society for the Study of Amphibians and Reptiles

Osteoderms in Anurans
Author(s): Rodolfo Ruibal and Vaughan Shoemaker
Reviewed work(s):
Source: Journal of Herpetology, Vol. 18, No. 3 (Sep., 1984), pp. 313-328
Published by: Society for the Study of Amphibians and Reptiles
Stable URL: http://www.jstor.org/stable/1564085 .
Accessed: 04/07/2012 08:32
Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at .
http://www.jstor.org/page/info/about/policies/terms.jsp

.
JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of
content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms
of scholarship. For more information about JSTOR, please contact support@jstor.org.

Society for the Study of Amphibians and Reptiles is collaborating with JSTOR to digitize, preserve and extend
access to Journal of Herpetology.

http://www.jstor.org

Journalof Herpetology, Vol. 18, No. 3, pp. 313-328, 1984

Copyright 1984 Society for the Study of Amphibians and Reptiles

Osteoderms in Anurans
RODOLFO RUIBAL AND VAUGHAN SHOEMAKER
Department of Biology, University of California,Riverside,California92521, USA
are more common in anurans than has been genABSTRACT. - Dermal ossifications-- osteodermserally acknowledged. In the hylid Phyllomedusa bicolor osteoderms are located in the dermis, cover
the dorsal surface of the head and body, and are scattered on the lateral and ventral surfaces and
the limbs. Each osteoderm consists of a vascularized bony basal plate (0.1 mm thick) from which
bony lamellar spines protrude into the epidermis. The dorsal body osteoderms are approximately
3 mm2 in area. Similar osteoderms are present in Phyllomedusa vaillanti and Gastrotheca weilandii.
In the pelobatid Megophrys nasuta osteoderms are present in the dorsal body skin and although
comparable to the osteoderms of Phyllomedusa in size and shape they are histologically very different. Megophrys osteoderms are avascular and composed of calcified collagen bundles in an
orderly three-dimensional arrangement. The leptodactylid Hylactophryne augusti also has small
bony osteoderms in the skin of the dorsum. They differ from the hylid osteoderms and resemble
Megophrys osteoderms in being avascular and having a matrix composed of horizontal and vertical
bundles of collagen.
Larger dermal bony dorsal plates are present in Lepidobatrachus and Ceratophrys (Leptodactylidae) and in Brachycephalus (Brachycephalidae).
Anuran osteoderms are structurally different from, and not homologous with, caecilian dermal
scales. Given the histological differences among the various anuran osteoderms and the taxonomic
diversity (Hylidae, Pelobatidae, Leptodactylidae, and Brachycephalidae), osteoderms appear to have
been independently evolved a number of times within the Anura.
It is suggested that the term dermal scale be restricted to the bony scale of fishes and caecilians,
and osteoderms be used to denote the dermal scutes of anurans and reptiles.

Textbooks on vertebrate anatomy

clearly state that amphibians, with the
exception of the caecilians, are devoid
of scales (Romer and Parsons, 1978;
Smith, 1960). Reif (1982), in his review
of the dermal skeleton of vertebrates,
also states that among the Amphibia
only the caecilians "have a squamation
of bony scales." The herpetological literature also supports this view (Porter,
1972; Noble, 1931; Goin et al., 1978).
Careful reading of the literature indicates that there is confusion about the
definition of a scale. In the case of fish
scales it is defined as a bony (or collagenous) dermal scute and is usually referred to as a dermal scale. In contrast,
reptilian scales are keratinous epidermal outgrowths. However, reptiles are
also considered to possess dermal scales
in the form of bony plates, or osteoderms (Romer, 1956). In his Dictionary
of Herpetology, Peters (1964) refers to
the scales of caecilians as osteoderms.

Smith (1960) defines the bony dermal

scales of all tetrapods as osteoderms. The
confusion arises in the case of a number
of anurans that have isolated integumentary ossifications. For example,
Boulenger (1882) describes "bony deposits" in the dorsal skin of Phyllomedusa bicolorand P. vaillanti. He also states
that the dorsal skin of Megophrys nasuta
has "bony deposits" analogous to the
dorsal shield of Ceratophrysand mentions the bony dorsal plates of Brachycephalus. Noble (1931) acknowledges
that "secondary deposits of bone" occur
in the dorsal skin of some anurans and
mentions that Gastrotheca weilandii is
studded with "calcareous plates." More
recently Funkhouser (1957) refers to the
"osteoderms" found in the dorsal skin
of various species of Phyllomedusa, and
Lynch (1971) describes the dorsal armor
of Ceratoprhysand Lepidobatrachusas a
"loosely consolidated plate of osteoderms."

314

R. RUIBALAND V. SHOEMAKER

It would appear from the literature

that some anurans do have dermal
"scales" or osteoderms-despite
the
often stated remarks to the contrary. Of
course this depends on how we define
a scale. In this study the histology and
morphology of some of these anuran
dermal ossifications are examined with
the intent of clarifying the status of dermal scutes in amphibians.

California Academy of Sciences, San

Francisco. Voucher specimens of cleared
alizarin stained material have been deposited at these museums in the departments of herpetology.
RESULTS

Phyllomedusabicolor
Structure of Osteoderms.-Live specimens of Phyllomedusa bicolor do not
AND METHODS
MATERIALS
demonstrate any evidence of osteoderms
when handled or observed with
from
were
obtained
skin
of
Samples
the
naked
Matefresh
and
eye. Under the dissecting mispecimens.
preserved
rial for light microscopy was fixed in croscope the dorsal skin of live P. bicolor
Bouin's or Zenker's solution for decal- shows small isolated pink areas, about
cification and embedded in paraffin, 0.1 mm apart, that contrast with the sursectioned at 10 Atm,and stained with he- rounding green of the chromatophores.
matoxylin and eosin, Mallory's triple The epidermis is colorless and transparstain, or alcian blue (Humason, 1962). ent. Scanning electron micrographs of
Skin samples or entire specimens were the dorsal skin of P. bicolor show that
also cleared and stained with alizarin the pink areas are actually protruding
red S. The material for scanning elec- spines covered by epidermis (Fig. 1).
tron microscopy (SEM) was fixed in for- The epidermis is intact over most of the
malin when the epidermis was re- spines. Vertical sections of the skin
tained, or macerated in a solution of show well developed spines about 0.15
trypsin to digest the soft tissue and al- mm high arising from basal plates lolow visualization of the bone surface. cated in the lower dermis (Fig. 2A). The
Some osteoderms were also exposed epidermis covering the spines is devoid
briefly to sodium hypochlorite. Hori- of the underlying pigment cells (Fig. 1B)
zontal and vertical ground sections of and consequently, in the living skin, the
dry bone were also prepared. This per- spines appear pink due to the reflection
mitted the visualization of the osteo- of light from the nonpigmented deeper
cyte lacunae. Scanning electron micros- portions of the dermis.
Histological examination confirms
copy was done with a JEOLJSM 35C at
15 Kv and the samples were sputter that the spines and basal plates are comcoated with gold. P. bicolorskin and os- posed of bone. The material shows a lateoderms were also fixed in formalin, mellar structure and isolated osteocytes.
embedded in Spurr's resin and sectioned Ground dry sections of the plates demat 1 ,tm and stained with methylene onstrate characteristic osteocyte lacunae
blue. Skin of living animals was also with radiating canaliculi, with most of
studied under a dissecting microscope. the lacunae in concentric or linear patLive specimens of Phyllomedusabicol- terns. Ground sections viewed with the
or,.Osteopilus septentrionalis,and Mego- SEM show the characteristic lacunae of
phrys nasuta were purchased from com- bone (Fig. 2B). Much of the bone is parmercial dealers. Preserved specimens of allel fibered and lamellae can be disother species were obtained from the cerned in the spines (Fig. 1B) and on
Museum of Natural History at the Uni- the basal plates. Observed under a poversity of Kansas; Field Museum of Nat- larizing microscope, adjacent layers
ural History, Chicago; Natural History have similar fiber orientation. Portions
Museum of Los Angeles; Smithsonian of the basal plates can be characterized
Institution, Washington, D.C.; and the as woven bone, showing an irregular

OSTEODERMS

IN ANURANS

315

FIG.1. Phyllomedusabicolor.(A) Scanning electron micrograph of the surface of the dorsal skin. The
protruding spines are covered by the epidermis. The broken epidermis on some spines appears to be
an artifact of preparation. (Scale = .05 mm) (B) Vertical section of the tip of an osteoderm spine. The
thin and stretched epidermis covers the tip and no pigment underlies the epidermis in this area. Note
the lamellated appearance of the bony tip. Hematoxylin and eosin. (Scale = 25 Am) (C) Scanning
electron micrograph of the surface of an osteoderm. The sharply tapered spines arise from the basal
plate of the osteoderm. (Scale = 0.1 mm)

316

R. RUIBAL AND V. SHOEMAKER

FIG.2. Phyllomedusa
bicolor.(A) Verticalsection of the dorsalskin. The more basophilicspine appears
darkerthan the basalplate. Vascularcanalsare present in the basalplate. The epidermisover the spine
is complete and stretchesover the protrudingtip of the spine. Hematoxylinand eosin. (Scale= 50 Am)
(B) Lacunaof an osteocyte. Scanningelectron micrographof the ground surfaceof a dorsalosteoderm.
(Scale = 5 Am)

distribution of osteocytes and fibers

(Pritchard, 1972). Vascular canals are
present in the basal portion and spines
(Fig. 2A), and connect with the surrounding looser connective tissue by
lateral and vertical foramina. Some of
the vascular canals show concentric lamellae of bone around the lumen. A
periosteal layer can be seen in some sections.
In decalcified sections, the bony matrix is eosinophilic with some basophilic staining of the tips of the spines. Alcian blue stains only the tips of some
spines.
We specifically searched for evidence
of a dentine or enamel-like outer surface on the osteoderms. Neither the 10

,um decalcified sections nor the thin (1

,um)vertical sections of resin-embedded
undecalcified osteoderms showed evidence of surface differentiation. Dry
ground vertical sections, studied with
SEM, also failed to reveal a distinct outer surface on the spines or basal plate.
Distributionof Osteoderms.-The entire
dorsal surface of the body and head of
P. bicolor is covered by irregularly
shaped bony plates, closely juxtaposed
and with indented borders (Fig. 7A).
The osteoderms of the body vary in size
and are usually greater than 1 mm in
their longest dimension with an area of
approximately 3.0 mm2. The osteoderms
are pierced by vertical foramina and the
outer surface studded with spines (about

IN ANURANS
OSTEODERMS

FIG.3. Alizarin-stained osteoderms of Gastrotheca weilandii. The space between the individual
osteoderms is well demarcated. (Scale = 0.5 mm)

0.15 mm in height) and low ridges (Figs.

1C and 7A). The basal portion is about
0.1 mm thick. Over the head, individual
osteoderms are not visible and seem to
form a single fused plate with larger
vertical perforations. The osteoderms of
the head are free from the underlying
skull bones and there is no evidence of
coossification as found in Osteopilus
(Trueb, 1966). On the lateral surfaces of
the body the osteoderms grade into
smaller (<0.5 mm) isolated plates that
are widely separated. Patches of similar
isolated osteoderms are present on the
ventral surface in the pectoral and gular regions. These small osteoderms reside at the base of the stratum spongiosum and just above the denser
stratum compactum. A few small osteoderms are also present on portions of
the fore and hind limbs.
Other Species
Phyllomedusa.-Dorsal skin samples
from seven other species of Phyllomedusa (sauvagei, hypochondrialis, azurae,
perinesos, tarsius, tomopternaand vaillanti) were examined by clearing and staining with alizarin. Osteoderms were
found only in P. vaillanti. The individ-

317

ual osteoderms are as large as in P. bicolor but differ in having larger perforations, angular spiculate margins, and
a lower total surface occupied by bone.
Gastrotheca.-Alizarin stained dorsal
skin samples from G. weilandii (Fig. 3)
demonstrated well developed osteoderms resembling the dorsal body osteoderms of P. bicolor.Ground horizontal sections of dry osteoderms showed
numerous concentrically and linearly
oriented osteocyte lacunae with abundant canaliculi. Most of the osteoderms
showed a fine lamellar structure. The
osteoderms are approximately 1 mm in
length and have an area of 1 mm2. Dorsal skin samples from G. longiceps and
G. riobambaeshowed no evidence of osteoderms.
Megophrys.-M. nasuta has well developed, irregularly shaped, and tightly juxtaposed osteoderms in the anterior dorsal body skin. The osteoderms
are 1 to 2 mm in length (0.5-2.5 mm2)
and extend as a dorsal band from just
posterior to the skull to the sacrum.
Vertical sections of decalcified dorsal
skin show spines arising from basal
plates (Fig. 4A). The spines do not protrude into the epidermis and have a
more rounded apex (Fig. 5) than in
Phyllomedusa. The main mass of the osteoderm stains with eosin and the outermost surface is basophilic. The basal
plate shows a lamellar structure made
up of horizontal layers of fibers. There
are also vertical bundles of fibers (Sharpey's fibers) extending between the inner and outer surface of the osteoderm,
most clearly seen with the polarizing
microscope. The cells reside in rounded
lacunae and appear to have pycnotic
nuclei. With alcian blue the osteoderm
is unstained except for the outermost
surface. The osteoderms lack any evidence of vascular canals in the spines
and basal plates. The ventral portion of
the basal plates grades into large (8 ,m
diameter) collagen bundles layered at
right angles to each other.
Under the light microscope horizontal ground sections of dry osteoderms

R. RUIBAL AND V. SHOEMAKER

318

I-

r
rq
t~~7;"
~~~~~~~~~L~~~~~~~
~~,
4 "

i", 'T~~~~~C
mwiv-~ 'tB~

. ,

~?l~iL~~
. ... rL~"?~ 1~~~~~~~~~~~"L??l*
1~

FIG.4. Megophrys nasuta. (A) Vertical section through the dorsal body skin. The blunt spines of the
osteoderms do not protrude into the epidermis and a well formed layer of loose connective tissue
resides between the osteoderm and the epidermis. The underlying dense connective tissue shows a
lamellated structure that grades into the lower surface of the osteoderm. Vertical bundles of collagen
(Sharpey's fibers) are also present. No blood vessels are present in the osteoderm. Hematoxylin and
eosin. (Scale = 0.1 mm) (B) Scanning electron micrograph of the horizontally ground surface of a dry
osteoderm. Bands of horizontal collagen fibers, at right angle to each other, give a parquet-like pattern.
Vertical collagen bundles (Sharpey's fibers?) are in cross section and appear whitish. The large empty
foramen is probably one of the vertical canals that traverses the osteoderm. (Scale = 0.1 mm)

show osteocyte lacunae with canaliculi,

but the lacunae are not arranged in any
pattern. Viewed with the scanning electron microscope the horizontal ground

sections show a parquet-like pattern of

horizontal fibers at right angles to each
other as well as cross sections of the
vertical fiber bundles (Fig. 4B). The ver-

IN ANURANS
OSTEODERMS

FIG. 5. Scanning electron micrograph of the

surface of a Megophrys nasuta osteoderm. The
spines are blunter and shorter than in Phyllomedusa bicolor.(Scale = 0.1 mm)

tical fibers may represent the continuation of the Sharpey's fibers (Fig. 4A).
alizarin
Hylactophryne.-Cleared,
stained dorsal skin of H. augusti demonstrates polygonal osteoderms, tightly
juxtaposed, varying in size from 0.2-0.8
mm2. Vertical sections demonstrate osteoderms composed of a basal plate (0.1
mm thick) with rounded projections on
the outer surface not extending to the
epidermis (Fig. 6). The loose connective
tissue between the osteoderms and the
epidermis is well formed and contains
numerous glands. The decalcified osteoderm is basophilic, with portions of
the outer surface intensely basophilic.
These same areas stain with alcian blue.
The basal plate shows horizontal lamellae as well as vertical bundles of fibers
extending through the entire thickness
of the bone. Osteocytes are visible but
no vascular canals are present in the osteoderms. Below the osteoderms there
is a layer of dense connective tissue
(stratum compactum) with horizontal
layers of collagen fibers and with ver-

319

tical bands of collagen extending between and into the individual osteoderms. Ground horizontal sections show
abundant, small osteocyte lacunae in no
discernible pattern. SEM of the surface
of the osteoderms (Fig. 7C) shows a pattern very different from that of P. bicolor. The surface rugosities are concentrated in the central portion of each
osteoderm and the margins have pronounced slot-like perforations between
adjacent osteoderms.
In contrast to Phyllomedusa and Megophrys the osteoderms of Hylactophryne
are well developed on the lateral and
ventral surfaces of the body.
Lepidobatrachus.-Sections of the dorsal shield of a preserved adult specimen
of L. llanensis revealed a 0.5 mm thick
bony shield with epidermis and loose
connective tissue overlying the bone.
The shield is well vascularized, with
large cavities containing blood vessels.
The decalcified matrix is eosinophilic
and, except for patches of the outermost
surface, fails to stain with alcian blue.
Most of the tissue around the cavities
and the surface has a lamellar structure,
but much of the matrix seems to be a
woven bone (Pritchard, 1972) with an
irregular arrangement of the fibers and
lacking a linear arrangement of the osteocytes. However, ground horizontal
sections of dry bone show typical osteocyte lacunae and a linear arrangement
of some of the lacunae.
Ceratophrys.-Decalcified vertical sections of the lateral edge of the dorsal
shield of a preserved specimen of C.
cranwelli revealed a bony structure similar to that of Lepidobatrachus.The bone
contains large vascular canals enclosed
by thin lamellae that resemble secondary ossification. Most of the material of
the shield is eosinophilic, but the apices
of the dorsal rugosities are basophilic,
stain with alcian blue, and show fine
lamellae. The outer and inner surfaces
of the shield are lamellar bone while
most of the main body of the shield
demonstrates a coarser woven bone or
regular alternating bands of large hor-

320

R. RUIBAL AND V. SHOEMAKER

:4.
~~~~.- ~

'

'~~~

't-i

Ilk~~~44

FIG. 6. Vertical section through the dorsal body skin of Hylactophryne augusti. The layer of loose
connective tissue between the osteoderms and the epidermis is well developed and contains relatively
large glands. The layered dense connective tissue below the osteoderms has horizontal collagen bundles with fibers in alternating orientation as well as vertical fibers. No blood vessels are present in
the osteoderms. Note the basophilic outer layer of the osteoderm. Hematoxylin and eosin, 10 im
section. (Scale = 50 Mm)

izontal collagen bundles as well as vertical fibers. Ground horizontal sections

of portions of the dorsal shield show
well developed osteocyte lacunae, often
linearly arranged.
were obBrachycephalus.-Samples
tained from museum specimens of B.
ephippium. The number and arrangement of the dorsal body plates vary be-

tween individuals (Cochran, 1955).

There are two large lateral plates that
are fused medially to the laterally expanded neural spines of the vertebrae,
and smaller plates anterior and posterior to the large lateral plates. Medial
plates are fused to the flattened apex of
the neural spine.
Vertical sections of decalcified lateral

OSTEODERMS IN ANURANS
5

A'

. ,&I

321

_
X -..

V.

,w

.*

*4LV;7
If2t^J
,^

't-7-1

&?
I.*
'*

..
I
Sl

AL
i
*.

I
)a

It

..a
tI*.I
FIG.7. Scanning electron micrographs of dorsal body osteoderms. (A) Phyllomedusabicolor.(B) Megophrys nasuta. (C) Hylactophryne augusti. The osteoderms were cleaned by digestion in trypsin and

brief exposure to sodium hypochlorite. The margins between the individual osteocytes are visible as
slot-like perforationsare
well as the vertical foraminathat perforatethe osteoderms.In Hylactophryne
present between adjacent osteoderms. Similar, but less pronounced, slot-like perforationsare also
present in Megophrys.(Scale = 0.5 mm)

322

R. RUIBALAND V. SHOEMAKER

plates have a thickness of about 0.2 mm

and show rounded rugosities on the
outer surface that in some cases displace
the loose connective tissue and lie adjacent to the epidermis. The inner surface of the plate may have a very narrow band of dense connective tissue or
be in direct contact with the underlying
muscle (Fig. 8). The plate has large vascular canals with well marked concentric lamellae surrounding the lumen.
Other portions show a pattern of horizontal lamellae and cells are visible between the fibers but no linear pattern
is evident. Ground horizontal sections
show well developed and abundant osteocyte lacunae without an apparent
pattern to their distribution.

derms as "bony deposits" and thus

avoided the problem of having to determine whether they constituted structures that could be homologized with
the dermal scales of caecilians and fishes or the osteoderms of reptiles. Gadow
(1901) stated that dermal scales in amphibians are restricted to caecilians but
added that "calcifications" or "calcareous deposits" were present in some
anurans. Without justification he concluded that these few anuran dermal
structures did not justify being considered osteoderms. Others have avoided
the term osteoderm in describing the
dermal armor of anurans, i.e., Inger
(1966) refers to Megophrysas having the
"skin behind the head calcified."
The osteoderms of anurans are very
Developmentof Osteoderms
different from the scales of caecilians.
Dorsal skin samples from three juve- In caecilians the scales are flat, ovoid,
nile specimens of Phyllomedusavaillanti and usually located in groups of three
(SVL = 21, 27 and 35 mm) were cleared or more scales in deep pockets in the
and stained with alizarin. The 21 mm dermis (Wake, 1975; Zylberberg et al.,
specimen was recently metamorphosed 1980; Fox, 1983). The scales are obliqueand had a nubbin of tail tissue. Neither ly oriented with the basal portion loof the two smaller specimens showed cated most anteriorly. Vertical sections
any evidence of osteoderms. The 35 mm of the scales demonstrate a number of
specimen had small isolated osteo- layers including an outer surface of
derms having a mean length of approx- mineralized plates (squamulae). The
imately 0.2 mm. In contrast an 80 mm deeper layers of the scale are fibrous
SVL specimen had plates greater than 1 (collagenous) and cellular. Zylberberg
mm in length.
et al. (1980) describe caecilian scales as
A 30 mm SVL juvenile specimen of comparable to teleost scales, but very
Ceratophrysornatawas sectioned and did different from the osteoderms of repnot show any evidence of a dorsal tiles. Scanning electron microscopy of
shield. This confirms Lynch's (1971) the bony surface (Perret, 1983; Casey
statement that juvenile ceratophryids and Lawson, 1979; Zylberberg et al.,
lack a dorsal shield. Cleared, alizarin 1980; Fox, 1983) show an ornamented
stained larvae and recently metamor- surface with distinct squamulae demarphosed specimens of Lepidobatrachus cated by unossified furrows. Caecilian
laevis did not show any evidence of a scales may thus be accepted as direct
dorsal shield.
homologues of fish scales-a primitive
character retained by many caecilians.
DISCUSSION
The scales of caecilians resemble those
Dermal ossifications-osteodermsof the Permian temnospondyl Trimerorclearly are present in anurans. In spite hachis.Olson's (1979) description of the
of their mention by Boulenger (1882), latter (which differs from that of ColNoble (1931) and Funkhouser (1957) the bert, 1955) is of small 1-7 mm oval scales
existence of osteoderms in anurans gen- situated deep in the dermis, with surerally has been ignored. Both Boulen- face ornamentation, and in overlapping
ger and Noble referred to the osteo- groups of 3 to 20 scales.

IN ANURANS
OSTEODERMS
The only resemblance between caecilian scales and anuran osteoderms is
that both are dermal and bony. The implication is that anuran osteoderms are
new structures. They are related to other dermal ossifications only in their
presumed origin from the interaction of
mesenchyme with the epidermis.
In reptiles, osteoderms are common
in several lizard families, turtles and in
the Crocodylia. Moss (1969) compared
the histology of various reptilian osteoderms and concluded that there was
great diversity and no clear morphological relationship between the osteoderms of recent reptiles. Like Romer
(1956), he concluded that reptilian osteoderms were new structures and not
descendant from fish scales. Lizard osteoderms are associated with the overlying keratinous scales. The osteoderms
may form single ossifications or be composed of individual plates and are in
the deeper dermis with loose connective tissue between the epidermis and
the bone. Moss emphasized the histological diversity of lizard osteoderms
and implied that osteoderms may have
arisen more than once in lizards. However, Moss (1968) also concluded that
functionally "all dermal skeletons are
homologous." However, this is not an
appropriate or useful concept of homology (Gans, 1969).
There are apparently two forms of
dermal dorsal armor in anuransclosely juxtaposed small osteoderms
(Phyllomedusa, Gastrotheca, Megophrys
and Hylactophryne) and larger, thicker
bony plates (Ceratophrys, Lepidobatrachus, and Brachycephalus).All of these
examples of anuran dermal armor are
composed of bone. The evidence for this
is that all stain with alizarin red S, the
mineral component of the armor is dissolved in acid, the matrix appears to be
collagen, and, most importantly, all
show the presence of osteocytes (lacunae with canaliculi). The histology of
bone can be varied and difficult to interpret (Hall, 1975, 1978; Moss, 1969).
Moss used the phrase "dermal sclerifi-

323

cation" to describe the reptilian osteoderms because he was unable to reconcile their diverse histology with the
conventional definition of bone. In
contrast, the anuran osteoderms studied
so far can all be classified as bone.
Phyllomedusa, Gastrotheca,Brachycephalus, Ceratophrys, and Lepidobatrachus
show fairly typical bone-thin
lamellated and concentric layers of collagen
with some of the osteocytes in linear
and/or concentric patterns, and with
vascular canals perforating the tissue.
In contrast Megophrys and Hylactophryne show a complex but orderly pattern of thick collagen bundles arranged
at right angles to each other, and have
abundant osteocytes, without any linear pattern to the arrangement of the
lacunae. Furthermore, the osteoderms
of Hylactophryne and Megophrys are
avascular. The osteoderms of Hylactophryne and Megophrysare similar to each
other, but only the Megophrys osteoderms have been studied in detail.
Scanning electron microscopy, and ordinary light and polarized light microscopy, all indicate a unique orderly array
of collagen bundles-a
three dimensional arrangement of fibers at right angles to each other (Fig. 4B). The lack of
blood vessels or a secondary lamellated
structure, and the appearance of the
cells, sharply distinguish them from the
dermal ossifications of Phyllomedusa.
Enlow (1969) describes avascular bone
in reptiles. However, the histology of
reptilian avascular bone does not resemble that of Megophrys osteoderms.
We know little of the development of
anuran osteoderms. In Phyllomedusa
vaillanti no osteoderms are present at
metamorphosis or soon after. In Lepidobatrachuslaevis and Ceratophrysornata
the dorsal shield is not present in young
postmetamorphic juveniles. Dermal ossification of the skin thus appears to develop late in postmetamorphic life. It
may be assumed that the osteoderms
first appear as isolated dermal ossifications that enlarge with time, and at least
in the dorsal skin, grow to abut. Evi-

324

R. RUIBALAND V. SHOEMAKER

dence of partially fused sutures was seen

in some P. bicolorosteoderms.
The large thick ossifications of the
ceratophryds and Brachycepahlusoccupy
most of the subepidermal space of the
integument. There is a thin layer of
loose connective tissue (stratum spongiosum) between the bone and the epidermis, but the bone replaces most of
the deeper stratum compactum characteristic of anuran skin. In Ceratophrys
and Lepidobatrachus
the stratum compactum is usually present only along the
lateral portion of the dorsal shield. In
the unossified dermis, lateral to the
bony shield, the horizontal fibers of the
stratum compactum terminate in direct
contact with the bony shield. In both
Brachycephalus and the ceratophryds
much of the dorsal armor lies directly
over somatic musculature (Fig. 8). In
contrast the basal plates of the smaller
osteoderms of Phyllomedusa,Gastrotheca,
Megophrys, and Hylactophrineare located between the outer loose connective
tissue and the deeper and denser stratum compactum. The collagen fibers of
the stratum compactum are often continuous with the ossified lower surface
of the osteoderm (Figs. 4a and 6).
Elkan (1968 and 1976) has described
a "ground substance" (GS) layer in the
dermis of anurans. In some species the
GS layer is situated between the stratum compactum and the stratum spongiosum. Elkan has claimed that this layer is high in acid mucopolysaccharides
and calcium, since in some species the
layer has been shown to stain with alcian blue and alizarin. It is thus possible
that this may be the site for the initial
ossification of the osteoderms.
Dermal ossifications in anurans are
located primarily in the anterior dorsal
body skin and cover the body cavity and
spinal cord. It thus appears as if the osteoderms represent protective dermal
armor. Another possible function would
be water conservation, as has been described for the coossified integument
and skull bones of some hylids such as
Osteopilus (Trueb, 1966; Seibert et al.,

1974). However, this would be a poor

argument in the case of Phyllomedusabicolor since, like other phyllomedusines,
this species secretes skin wax (personal
observation) and consequently has a
more effective mechanism for reducing
water loss (Shoemaker and McClanahan, 1975). Furthermore, in most
of the armored species the tissue between the osteoderms and the epidermis is richly supplied with blood vessels.
The ecology of these armored species
is very diverse. Phyllomedusabicolorand
P. vaillanti are nocturnal arboreal frogs
of the Amazonian forest. Gastrothecainhabits the wet tropical forests of South
America. The species of Lepidobatrachus
are aquatic, inhabiting ephemeral pools
in arid and semi-arid portions of Argentina and Paraguay. Ceratophrys
species are more terrestrial and found
in semi-arid and mesic areas of South
America. During the dry season Lepidobatrachusand Ceratophrysare fossorial
and become enclosed in an epidermal
cocoon. In contrast Megophrys nasuta is
terrestrial, living in the leaf litter of the
forests of southeast Asia and Malaya.
Hylactophryneaugustiis a terrestrial rock
dwelling species associated with limestone outcroppings in Mexico and
southwestern United States. Brachycephalusis diurnal and inhabits the forest floor in southeastern Brazil. These
species have little in common ecologically that would help indicate the functional value of the dorsal armor.
The various dermal ossifications of
anurans are morphologically distinct
from the dermal scales of fishes and
caecilians, and lack the association with
keratin scales characteristic of reptilian
osteoderms. If we accept the definition
of an osteoderm as a dermal ossicle-a
bony plate in the skin-then anurans
have osteoderms. The dermal scales of
fishes and caecilians seem to be homologous as a result of descent from an ancestral dermal scale. However, the dermal ossifications of anurans clearly are
not homologous with caecilian scales.

OSTEODERMS IN ANURANS

325

FIG.8. Vertical section of the dorsal skin and armor of Brachycephalus

The lamellated
ephippium.
structureof the bone and the bony projectionin contact with the epidermis is visible. Hematoxylin
and eosin. (Scale = 50 Am)

Both the structure of the osteoderms and

their scattered distribution within anuran families (Hylidae, Pelobatidae,
Leptodactylidae, and Brachycephalidae) argue against such a homology. Instead it appears as if osteoderms have
been repeatedly evolved de novo. Certainly the diversity of osteoderms in
lizards (Moss, 1969) indicates that they

have arisen more than once in this

group. The fact that anuran osteoderms
also show histological diversity, are
found in a number of families, and are
only present in some of the species
within a genus argues for multiple evolutionary origins within the Anura.
Given the apparent lack of precision
in the use of the terms dermal scale and

326

R. RUIBALAND V. SHOEMAKER

osteodermsome attempt at clarification

seems appropriate. Krejsa (1979) distinguishes between fish dermal scales and
reptilian osteoderms on the basis of embryology. Fish dermal scales are presumed to be derivatives of an epidermal/ectomesenchyme interaction. The
dermal scales of fishes start as an aggregation of mesenchyme cells that form a
dermal papilla associated with an epidermal pocket. In contrast reptilian osteoderms are considered by Krejsa(1979)
to be exclusively mesodermal in origin.
Unfortunately we do not have direct
evidence that all fish scales are the result of epidermal/ectomesenchyme interaction or that the ectomesenchyme
(neural crest cells) actually contribute
to fish scales. In the case of caecilians,
Marcus (1934) describes the development of scales in Hypogeophisand states
that the scales are the result of ectodermal interactions with the underlying
mesoderm. As Reif (1982) points out,
this has not been confirmed (or denied)
by any workers since that time. Furthermore, we do not know whether the
varied reptilian osteoderms are formed
solely by the mesodermal mesenchyme.
Nevertheless it would appear theoretically worthwhile to restrict the term
dermal scale for the dermal scutes of
fishes and caecilians. This acknowledges the apparent homology of these
ossifications. The term osteoderm can
then be retained for the dermal ossifications of anurans and reptiles, but it is
essential to recognize that osteoderms
represent non-homologous structures
that have been independently evolved
a number of times. This classification is
close to that suggested by Smith (1960)
who denoted all tetrapod (including
caecilians) dermal bony scutes as osteoderms. Unfortunately our suggested
classification is based on a number of
assumptions about the epigenetic history of these dermal structures. Nevertheless the classification has the merit
that the assumptions are explicit and
consequently exposed to verification or
denial.

Moss (1969) suggested that some lizard osteoderms contain a ganoin-enamel-like outer surface. His evidence was
that the outer layer of the lizard osteoderms was basophilic and alcian blue
positive. The principal substances
stained by alcian blue are glycosaminoglycans (acid mucopolysaccharides)
which constitute the extracellular
ground substances of connective tissue
(Cook, 1982). The anuran osteoderms
show alcian blue staining and basophilia at the tips of the spines of Phyllomedusa bicolor and portions of the outer
surface of the osteoderms of Hylactophryne and Megophrys. In P. bicolor,
which was studied in more detail than
the other species, no structural differences were seen in the spines. Furthermore, the lamellae of the spines (Fig.
1B) are seen to be continuous with the
lamellae of the basal plate and thus do
not represent an independent capping
of the tip. In Hylactophryne the alcian
blue region forms a band, 12-24 Am
thick, along the outer surface of the basal plate and the protuberances (Fig. 6).
The band is not dense, but rather has a
fibrous appearance. The Hylactophryne
sections were prepared from previously
preserved museum specimens. In Megophrys the alcian blue layer is similar
to Hylactophryne but has a denser appearance and on some of the protuberances showed a thin hyaline blue layer.
Both the basophilia and alcian blue
staining could result from an increase
in the density of the extracellular matrix. Polarized light microscopy did not
reveal surface birefringency of the outer layer of the osteoderm in any of the
species. For the present it would appear
prudent to interpret the alcian blue layer of anuran osteoderms as a region of
more recent bone deposition. Certainly
we have no evidence to contradict Reif's
(1982) contention that in tetrapods neither dentine nor enamel are found outside of the mouth.
Anuran osteoderms may prove to be
more common than is currently demonstrated. Funkhouser (1957) indicated

OSTEODERMS IN ANURANS

that in Phyllomedusa osteoderms were

present not only in bicolor and vaillanti
but also in orcesi (now P. tarsius according to Duellman, 1974) and possibly in
burmeisteri. Boulenger (1882) and Gadow (1901) also reported that Hyla dasynota (currently in the synonymy of
Hyla senicula) from Brazil had bony deposits in the skin from the skull to the
sacrum. Trueb (1973) mentions a small
dermal presacral dorsal shield in Dendrobates. We have been unable to confirm these reports.
Given the distinctness of the osteoderms of Phyllomedusa and Megophrys
and the fact that they were reported in
Boulenger's 1882 opus, it is surprising
that their presence has been ignored for
so long. Funkhouser (1957) is the only
author who clearly acknowledges their
prominence and her findings have been
all but ignored. That it has taken 100
years to rediscover and study in detail
what Boulenger described seems to be
the result of a conceptual fallacy. Since
caecilians were "known" to be the only
amphibians with dermal ossicles the
anuran "calcifications" were relegated
to a herpetological limbo where they
would not disturb an accepted paradigm.
Acknowledgments.-We are indebted
to many persons for their assistance
during this study. The loan of specimens was through the generosity and
interest of William Duellman and David Cannatella, Museum of Natural
History, University of Kansas; Ron
Heyer and George Zug, Smithsonian
Institution; John Wright and Robert
Bezy, Natural History Museum of Los
Angeles County; Robert Inger, Field
Museum of Natural History; and Robert
Drewes and Larry Wishmeyer, California Academy of Sciences. Tamir Ellis
and Randolph Wolff were eminently
successful in maintaining our captive
Phyllomedusa bicolor in good health.
Technical assistance in preparing samples for light microscopy and SEM, and
photographing material, was provided

327

by Marcia Kooda-Cisco, John Kitasako,

Betty Moore, Leah Haimo, Brian Gilpin,
and Spencer Kwong. Live specimens of
Megophrys nasuta and Ceratophrysornata
were provided by Claire and Steve Alt.
John S. Applegarth, who made the original discovery of osteoderms in Hylactophryne, very generously provided us
with information. Richard Estes, Richard Etheridge and Marvalee Wake read
and improved an earlier draft of this
article. We also thank Robert Drewes,
Ronald Crombie, and Norman Scott for
providing us with information about
some of the species studied. Supported
by NSF grant PCM-8302888.
Hobart Smith served as editor for this
article.
LITERATURE
CITED
BOULENGER,G. A. 1882. Catalogue of the Batrachia Salientia s. Ecaudata in the collection of
the British Museum. 2nd Ed., London. 503 pp.
CASEY,J., AND R. LAWSON. 1979. Amphibians with
scales: the structure of the scale in the caecilian Hypogeophisrostratus.Br. J. Herpetol. 5:831833.
COCHRAN, D. M. 1955. Frogs of southeastern
Brazil. U.S. Nat. Mus. Bull. 206:i-xvi, 1-423.
COLBERT,E. H. 1955. Scales in the Permian amphibian Trimerorhachis.Amer. Mus. Nov. 1740:
1-17.
COOK,H. C. 1982. Carbohydrates. In J. D. Bancroft and A. Stevens (eds.), Theory and Practice of Histological Technique, 2nd ed. Pp. 180216. Churchill Livingston, London.
DUELLMAN, W. E. 1974. Taxonomic notes on
Phyllomedusa(Anura:Hylidae) from the upper
Amazon basin. Herpetologica 30:105-112.
ELKAN, E. 1968. Mucopolysaccharides in the anuran defense against desiccation. J. Zool. London 155:19-53.
. 1976. Ground substance: an anuran defense against desiccation. In B. Lofts (ed.),
Physiology of the Amphibia 3. Pp. 101-110.
Academic Press.
ENLOW,D. H. 1969. The bone of reptiles. In C.
Gans (ed.), Biology of the Reptilia Vol 1, Pp.
45-80. Academic Press.
Fox, H. 1983. The skin of Ichthyophis(Amphibia:
Caecilia): an ultrastructural study. Jour. Zool.
London 199:223-248.
FUNKHOUSER,A. 1957. A review of the neotropical tree-frogs of the genus Phyllomedusa.Occ.
Pap. Nat. Hist. Mus. Stanford Univ. 5:1-90.
GADOW,H. 1901. Amphibia and reptiles. Cambridge Natural History 8:1-668.

328

R. RUIBAL AND V. SHOEMAKER

GANS,C. 1969. Discussion: Some questions and

problems in morphological comparisons. Ann.
N.Y. Acad. Sci. 167:506-513.
GOIN, C. J., O. B. GOIN, AND G. R. ZUG. 1978.
Introduction to herpetology. W. H. Freeman,
San Francisco. 378 pp.
HALL,B. K. 1975. Evolutionary consequences of
skeletal differentiation. Amer. Zool. 15:329-350.
- . 1978. Development and cellular skeletal
biology. Academic Press, New York. 304 pp.
G. L. 1962. Animal tissue techniques.
HUMASON,
W. H. Freeman. 468 pp.
INGER,R. F. 1966. The systematics and zoogeography of the Amphibia of Borneo. Fieldiana:
Zoology 52:1-402.
KREJSA,R. J. 1979. The comparative anatomy of
the integumental skeleton. In M. H. Wake (ed.),
Hyman's Comparative Vertebrate Anatomy.
3rd ed. Pp. 112-191. Univ. Chicago Press.
LYNCH, J. D. 1971. Evolutionary relationships,
osteology, and zoogeography of leptodactyloid frogs. Univ. Kansas Mus. Nat. Hist. Misc.
Publ. 53:1-238.
MARCUS,H. 1934. Beitrag zur Kenntnis der
Gymnophionen. Nr. XXI.Das Integument. Zeit.
f. Anat. u. Entw. 103:189-234.
Moss, M. L. 1968. Bone, dentine, and enamel
and the evolution of vertebrates. In P. Person
(ed.), Biology of the Mouth. Pp. 37-65. A.A.A.S.
Publ. 89, Washington, D.C.
. 1969. Comparative histology of dermal
sclerifications in reptiles. Acta Anat. 73:510533.
NOBLE,G. K. 1931. Biology of the Amphibia.
Dover Publ. 577 pp.
OLSON, E. 1979. Aspects of the biology of Trimerorhachis(Amphibia Temnospondyli). J. Paleont. 53:1-17.
PERRET,J. 1982. Les ecailles de deux Gymnophiones africains (Batraciens, apodes), observees au microscope electronique a balayages.
Bonn. zool. Beitr. 2-4:343-347.

PETERS,
J. A. 1964. Dictionary of herpetology.
Hafner Publ. Co., N.Y. 392 pp.
PRITCHARD,
J. J. 1972. General histology of bone.
In G. H. Bourne (ed.), The Biochemistry and
Physiology of Bone. Pp. 1-20. Academic Press,
London.
K. R. 1972. Herpetology. W. B. Saunders
PORTER,
Co., Philadelphia. 524 pp.
REIF,W. E. 1982. Evolution of dermal skeleton
and dentition in vertebrates. In M. K. Hecht,
B. Wallace, and G. T. Prance (eds.), Evolutionary Biology, Vol. 15. Pp. 287-368. Plenum Press,
N.Y.
A. S. 1956. Osteology of reptiles. Univ.
ROMER,
Chicago Press. 772 pp.
, ANDT. S. PARSONS.1978. The vertebrate
body. W. B. Saunders Co., Philadelphia.
AND R. J. WASE. A., H. B. LILLYWHITE,
SEIBERT,
SERSUG.1974. Cranial coossification in frogs:
relationship to rate of evaporative water loss.
Physiol. Zool. 47:261-265.
V. H., ANDL. L. MCCLANAHAN.
1975.
SHOEMAKER,
Evaporative water loss, nitrogen excretion, and
osmoregulation in phyllomedusine frogs. J.
Comp. Physiol. 100:331-335.
SMITH,H. M. 1960. Evolution of chordate structure. Holt, Reinhart, and Winston, New York.
529 pp.
TRUEB,L. 1966. Morphology and development
of the skull of the frog, Hyla septentrionalis.Copeia 1966:562-573.
. 1973. Bones, frogs, and evolution. Pp.
65-132 J. L. Vial (ed.), Evolutionary biology of
anurans. Univ. Missouri Press, Columbia.
WAKE,M. H. 1975. Another scaled caecilian
(Gymnophiona: Typhlonectidae). Herpetologica 31:134-136.
AND A. DERICQLES.
L., J. CASTANET,
ZYLBERBERG,
1980. Structure of the dermal scales in Gymnophiona (Amphibia). Jour. Morph. 165:41-54.
Accepted: 24 February 1984.