Veterinary Dermatology 2004, 15, 159 167

Blackwell Publishing, Ltd.

Canine distemper virus infection of canine footpad epidermis

ANDREA GRNE*, MARCUS G. DOHERR and ANDREAS ZURBRIGGEN
*Institut fr Tierpathologie and Departement fr Klinische Veterinrmedizin, Abteilung Klinische Forschung
Universitt Bern, Berne, Switzerland
(Received 7 June 2002; accepted 17 November 2003)

Abstract Infection of the footpad epidermis can occur in natural canine distemper virus (CDV) infection of
dogs. Footpads from 19 dogs experimentally inoculated with virulent distemper strain A75/17 and from two nonexposed dogs were examined histopathologically and assessed for the presence of viral antigen and nucleoprotein
mRNA, as well as number of inflammatory and apoptotic cells. Dogs were divided into four groups based on
inoculation status and postmortem examination: inoculated dogs with severe distemper (group 1, n = 7); inoculated
dogs with mild distemper (group 2, n = 4); inoculated dogs without distemper (group 3, n = 8); and noninoculated
dogs (group 4, n = 2). Footpads from dogs of all groups had a comparably thick epidermis. Eosinophilic viral
inclusions and syncytial cells were present in footpad epidermis of one dog of group 1. Footpads of group 1 dogs
contained viral antigen and mRNA in the epidermis with strongest staining in a subcorneal location. Additionally, in these dogs footpad dermal structures including eccrine glands and vascular walls were positive for virus
particles. No CDV antigen or mRNA was present in the footpad epidermis and dermis of any other dog. Group
1 dogs had more CD3-positive cells and apoptotic cells within the basal layer of the epidermis when compared
to the other groups. These findings demonstrate that in experimental infection CDV antigen and mRNA were
colocalized in all layers of the infected canine footpad epidermis. The scarcity of overt pathological reactions with
absence of keratinocyte degeneration indicates a noncytocidal persisting infection of footpad keratinocytes by CDV.
Keywords: canine distemper, dog, footpad, keratinocyte.

IN TRO D U CT I ON
Canine distemper virus (CDV), a morbillivirus, often
causes fatal disease in a variety of terrestrial mammals.1 Usually the gastrointestinal and respiratory
tracts are affected with or without involvement of the
nervous system.2 Infection of epidermal cells by CDV
or related morbilliviruses has been reported in different
species.37 Distemper in dogs is associated with a
vesiculo-pustular dermatitis of varying frequency.8
Another well known sequel, so-called hard pad disease,
is hyperkeratosis of the footpad.9 Involvement of the
footpad epidermis has been suggested to be common
in CDV infection and demonstration of viral antigen
has been proposed as an antemortem diagnosis.10
Changes in the footpad epidermis caused by CDV are
described as proliferation of basal keratinocytes and
orthokeratotic and parakeratotic hyperkeratosis, with
eosinophilic cytoplasmic inclusion bodies and vacuolar degeneration.11 However, in a study of dogs naturally infected with distemper virus, histopathological
changes in the footpad epidermis differed from this
description as neither inclusion bodies nor keratinocyte degeneration were significant characteristics.12
Histopathological evaluation of footpad epidermis from
This work was supported by the Swiss National Science Foundation
(grants #3158657.99 and # 32 61961.00).
Correspondence: A. Grne, Institut fr Pathologie, Tierrztliche
Hochschule Hannover, Bnteweg 17, 30559 Hannover, Germany.
E-mail: agroene@tiho-hannover.de
2004 European Society of Veterinary Dermatology

experimentally infected dogs yielded similar observations. The objective of this work was to analyse and
compare the localization of CDV antigen and mRNA
in these footpads. Additionally, inflammation and
apoptotic cells were characterized, counted, and correlated with presence of CDV antigen and mRNA.

M AT E R IA L S A N D M E T H O D S
The study population consisted of 21 juvenile,
5 month-old, specific pathogen-free Swiss beagle dogs
(11 male, 10 female), which were maintained according
to the guidelines of the institutions animal care and
use committee. Nineteen dogs (nos. 119) were inoculated with the virulent A75/17 CDV strain, which had
been propagated previously in dogs.13 Splenic extracts
(0.2 mL) from these dogs were inoculated via the vena
cephalica antibrachii. Dogs 20 and 21 were noninoculated animals originating from the same supplier.
Animals were monitored daily for clinical signs of
distemper. Any dog showing a decline in body condition or neurological signs was euthanized immediately.
All remaining dogs were euthanized with barbiturate
2036 days post inoculation and necropsied (Table 1).
Samples of body organs (brain, lung, stomach, kidney,
liver, heart, lymphatic organs, urinary bladder, intestinal tract and eyelids) were immersed and fixed in 4%
paraformaldehyde for 24 h, paraffin embedded and
cut. Sections were stained with haematoxylin and eosin
(H&E) for routine histopathology. Presence and
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A Grne et al.

Table 1. Number of dogs, days between inoculation and euthanasia, frequency of inclusions in various body organs, presence of CNS lesions
and presence of CDV in footpad epidermis in 21 beagle dogs after experimental inoculation with virulent A75/17 distemper virus
Viral inclusions
Group

Dog no.

Euthanasia
p.i. (days)

Lung

Stomach

Renal pelvic
epithelium

Urinary
bladder

1
1
1
1
1
1

1
2
3
4
5
6

20
24
24
28
28
32

Few
Few
Few
Few
Many
Few

None
Many
Many
Many
Many
Few

Many
Many
Many
Many
Few
None

Many
Many
Many
Many
Few
None

32

Few

Many

None

Few

31
31
33
33
3336
NA

Few
Few
Few
None
None
None

None
None
None
None
None
None

None
None
None
None
None
None

None
Few
None
None
None
None

2
2
2
2
3
4

8
9
10
11
1219
2021

Other organs
None
IT
Spleen, testis
Pancreas, IT
IT
Bile ducts,
meibomian glands
Tonsil, salivary
gland epithelium
IT
None
None
tonsil
None
None

CNS
lesions

CDV
(footpad epidermis)

Yes
Yes
Yes
Yes
Yes
Yes

Yes
Yes
Yes
Yes
Yes
Yes

Yes

Yes

No
No
No
No
No
No

No
No
No
No
No
No

IT, intestinal tract; NA, not applicable, as dogs were not inoculated.

number of viral inclusions were determined to assess

the severity of distemper. Dogs were divided into four
groups according to the necropsy findings (Table 1).
Group 1 included inoculated dogs with many inclusions or inclusions in more than two organs (severe distemper); group 2 included inoculated dogs with few
inclusions in 1 or 2 organs only (mild distemper);
group 3 included inoculated dogs without inclusions
in any organ, and group 4 were the noninoculated
dogs.
Metacarpal and metatarsal footpads were collected
at necropsy and processed as described. In order to avoid
interference with melanin during immunohistochemical
evaluation, nonpigmented metacarpal or metatarsal pads
were selected whenever possible. Sections (approximately 1 cm in length, one footpad per dog) were
stained with H&E for histopathological examination.
The presence of CDV antigen within the footpad epidermis and dermis was detected immunohistologically
using monoclonal antibody D110, which recognizes
CDV nucleoprotein.14,15 A positive reaction was visualized with 3-amino-9-ethylcarbazole (AEC, DAKO,
Zug, Switzerland), resulting in a red staining product,
after application of the secondary biotinylated antimouse antibody (DAKO). Staining intensity of CDV
antigen was judged as negative, weakly positive (a
granular light red) or strongly positive (intense red).
CDV nucleoprotein mRNA in the footpad epidermis
and dermis was identified using a digoxigenin-labelled
strand-specific RNA probe.16,17 A positive reaction was
visualized with nitroblue tetrazoliumchloride (NBT,
Boehringer Mannheim Biochemica, Mannheim,
Germany) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP, Boehringer Mannheim Biochemica),
resulting in a black-blue staining product. Staining
intensity for CDV mRNA was judged as negative,
weakly positive (light grey) and strongly positive (dark
grey to black).

Sections from the same footpads were also analysed

for the localization and numbers of macrophages
(lysozyme-positive cells), T-lymphocytes (CD3-positive
cells), and B-lymphocytes (CD79-positive cells).1820
The following antibodies were used: A099 (DAKO,
1 : 50) against lysozyme; A452 (DAKO, 1 : 50) against
CD3, and M7051 (DAKO, 1 : 40) against CD79.
Pretreatments were pronase (Fluka, Buchs CH, 1%,
pH 7.6 at 37 C for 5 min) for detection of lysozyme
and CD3-positive cells, and citrate (pH 6.0, 2 5 min
microwave at 750 W) for detection of CD79-positive
cells. Apoptotic cells were identified with an antiactivated caspase-3 antibody, kindly donated by
D. F. Montisano (Idun Pharmaceuticals, La Jolla, CA,
USA), in a dilution of 1 : 500. This antibody recognizes
the p18 subunit of cleaved caspase-3, but not the
unprocessed form.21 Positive immunohistochemical
reactions were visualized with AEC.
Inflammatory cells were identified and counted
separately in both the papillary zone of the footpad dermis and in the epidermis of adjacent rete ridges. Cells
positive for activated caspase-3 were counted in a similar
manner. Counts of 10 randomly chosen, different areas
per footpad containing the papillary zone and the rete
ridges using a 40 objective were added and the results
were compared between the four groups. Data were
analysed using NCSS 2001 (Number Cruncher Statistical Systems, Kaysville, UT, USA) and are represented as box and whisker plots with median (50th
percentile), interquartile range (IQR) and whiskers of
the length 75th percentile + 1.5 IQR and 25th percentile 1.5 IQR. Differences were tested for overall
statistical significance using a KruskalWallis one-way
on ranks (corrected for ties). For identification
of significant differences between individual groups the
KruskalWallis Z-test with Bonferroni adjustment for
multiple comparisons was used. The overall level of
significance was P < 0.05.

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Canine distemper virus in canine footpads

161

RESU LTS
Severe distemper was diagnosed in seven dogs (group
1) at postmortem examination (Table 1). These dogs
had many viral inclusion bodies in multiple organs
including lung, kidney, stomach and urinary bladder.
Additionally, these dogs had changes in the central
nervous system (CNS) typical for distemper (demyelination, inclusion bodies, astrocytosis). Four inoculated
dogs had mild distemper with individual inclusion
bodies in only one or two body organs (group 2). In
these dogs no changes indicative of distemper were
present in the CNS. Distemper was not diagnosed in
the remaining inoculated dogs (group 3) and was also
not present in the noninoculated dogs (group 4).
On histopathology, there was no apparent difference
in the thickness of the footpad epidermis, including the
stratum corneum, between the four groups of dogs.
Viral inclusion bodies were present in the footpad
epidermis of one dog of group 1 (Fig. 1). They were
located in the cytoplasm and nuclei of individual
keratinocytes in the stratum spinosum. Some of these
cells formed syncytia with multiple nuclei. Individual
keratinocytes exhibited vacuolar degeneration. Footpad
epidermis of some dogs had foci of degenerate or
necrotic keratinocytes. These foci were usually located
either at the epidermal-dermal junction or in the lower
layers of the epidermis. Such foci were more frequently
observed in footpads of group 1 dogs, and less commonly
or absent in footpads of the remaining dogs. The footpads
of most dogs of all groups occasionally had a mild perivascular infiltrate mainly composed of lymphocytes.
Viral antigen was consistently detected in keratinocytes of the suprabasal and subcorneal epidermal layers
of dogs 17 (Table 2). CDV nucleoprotein was present
in the vast majority of keratinocytes in these epidermal
layers and only rarely was viral antigen distributed in a
multifocal pattern (Fig. 2a). In dogs 2, 4 and 5, CDV
antigen was additionally observed in basal keratinocytes (Fig. 2b). There was distinct variation in staining
intensity. Strongest staining was present in the subcorneal layer (for details of distribution and intensity
see Table 2). In all dogs with CDV-positive footpads
CDV antigen was present in the endothelium and

Figure 1. Footpad epidermis from a dog with canine distemper

with intracytoplasmic eosinophilic inclusion bodies in keratinocytes
(dog no. 2). H&E, bar = 20 m.

pericytes and some fibroblasts; in four dogs CDV antigen

was additionally found in eccrine glands. All dogs with
CDV antigen in the footpad belonged to group 1.
CDV mRNA was present in the cytoplasm of
affected cells in footpads of seven dogs (nos. 17,
Table 2). Nucleoprotein transcripts were usually present in the epidermis in all layers, with the strongest
staining in subcorneal cells (Fig. 3a). Only in three
dogs (nos. 2, 4 and 5) did individual basal keratinocytes
contain viral mRNA (Fig. 3b). A distinct variation in
staining intensity was present depending on the location (Table 2). In the footpad dermis of dogs 15, CDV
transcripts were also present in epithelial cells of

Table 2. Intensity and distribution of CDV nucleoprotein and nucleoprotein mRNA in subcorneal, suprabasal, and basal layers of canine
footpad epidermis
Group 1
1
Nucleoprotein
Subcorneal
+
Suprabasal
+
Basal

Nucleoprotein mRNA
Subcorneal
+++
Suprabasal
+
Basal

Groups 24
2

821

++
+
+

++
+

+++
+
+

+++
+
+

+ +
+

+++
+

++
+
+

++
+

+++
+
+

+++
+
+

+
+

+++
+

Intensity of staining: , no positivity; +, weak positivity; + +, strong positivity.

Distribution of staining: , diffuse staining; , multifocal staining.
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A Grne et al.

Figure 2. (a) Canine distemper virus nucleoprotein in the epidermis

and individual cells of the dermis. Dog no. 2, immunohistochemistry,
avidin-biotin-peroxidase complex method. Bar = 80 m.
(b) Canine distemper virus nucleoprotein in basal keratinocytes
of the footpad epidermis (dog no. 5), immunohistochemistry,
avidin-biotin-peroxidase complex method. Bar = 20 m.

Figure 3. (a) Canine distemper virus nucleoprotein mRNA in

the epidermis and individual cells of the dermis (dog no. 2),
in situ hybridization. Bar = 80 m. (b) Canine distemper virus
nucleoprotein mRNA in basal keratinocytes of the footpad
epidermis (dog no. 5), in situ hybridization. Bar = 20 m.

eccrine glands, endothelial cells and pericytes, and

fibroblasts. All dogs with CDV transcripts belonged to
group 1. CDV mRNA and antigen was colocalized in
the footpad epidermis of affected dogs.
Inflammatory cells were seen in footpads of all dogs
to a small degree (Fig. 4). CD3-, CD79- and lysozymepositive cells were present in the papillary zone of the
superficial dermis. CD3-positive cells were the most
frequently observed inflammatory cells in the papillary

zone of the dermis. Group 1 dogs had significantly

more CD3-positive cells when compared to group 3
dogs but not when compared to the other groups
(Fig. 5). The number of CD79-positive cells was generally lower, and there was no significant difference in the
number of these cells between the four groups (Fig. 6).
Significantly more lysozyme-positive cells were present
in the papillary zone of group 1 dogs when compared
with group 2 and 3 dogs (Fig. 7). CD3-positive cells,

2004 European Society of Veterinary Dermatology, Veterinary Dermatology, 15, 159 167

Canine distemper virus in canine footpads

163

Figure 4. Inflammatory cells in canine distemper virus-positive

footpads (dog no. 3). (a) CD3-positive cells amongst keratinocytes.
(b) CD79-positive cells in the dermis. (c) Lysozyme-positive cells
in the dermis. Immunohistochemistry, avidin-biotin-peroxidase
complex method. Bars = 20 m.

Figure 5. CD3-positive cells in the dermis; box and whisker plots of

the total number of positive cells. Group 1 dogs had significantly
more positive cells compared to group 3 dogs.

Figure 6. CD79-positive cells in the dermis; box and whisker plots of

the total number of positive cells. There is no difference between the
numbers of positive cells of the four groups.

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A Grne et al.

Figure 7. Lysozyme-positive cells in the dermis; box and whisker

plots of the total number of positive cells. Group 1 dogs had
significantly more positive cells compared to group 2 and 3 dogs.

Figure 9. Cells positive for activated caspase-3 fragment p18 in the

epidermis; box and whisker plots of the total number of positive
cells. Group 1 dogs had significantly more positive cells compared to
the other groups.

Figure 8. CD3-positive cells in the epidermis; box and whisker plots

of the total number of positive cells. Group 1 dogs had significantly
more positive cells compared to the other groups.

but no CD79- and lysozyme-positive cells, were present

in the footpad epidermis. Also, group 1 dogs had significantly more CD3-positive cells in the footpad epidermis than the other groups (Fig. 8). Additionally, in
group 1 dogs these cells were often present in clusters
and not scattered individually as with the other dogs.
Apoptotic cells as identified by positive cytoplasmic
immunostaining for activated caspase-3 fragment p18
were rare or absent in footpad epidermis of dogs in
groups 24 (Figs 9 and 10). The footpad epidermis of
group 1 dogs had significantly higher numbers of apoptotic cells compared with the other groups. Apoptotic
cells were not seen in the papillary zone of the superficial dermis in any dog.

D ISCU SSIO N
Thickening of canine footpad epidermis in association
with an encephalitis (later identified as distemper) was

Figure 10. Activated caspase-3 fragment p18. Clusters of positive

cells in the epidermis (dog no. 7, immunohistochemistry, avidinbiotin-peroxidase complex method. Bar = 20 m).

first described by MacIntyre et al. (1948), and this disease complex was named hard pad disease.22 Similar
footpad lesions are reported to be seen in other diseases such as zinc-responsive dermatosis or pemphigus
foliaceus, and this may be one of the reasons why the
term hard pad disease is currently not used frequently.9 Histologically, this change is consistently
described as massive hyperkeratosis of the footpad epidermis.11,22 Additional features seen in CDV-infected
footpad epidermis include vacuolar degeneration of
keratinocytes, with formation of intracytoplasmic and
intranuclear inclusion bodies.11 These changes were
partly or totally absent in experimentally CDV-infected

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Canine distemper virus in canine footpads

footpads; therefore, presence of CDV particles was
examined and correlated with degenerative and inflammatory changes.
In this study, CDV antigen and mRNA was demonstrated in all layers of the epidermis. This could be the
sequel of cell-to-cell spread of the virus. Additionally,
potential movement of viral antigen with the outward
migration of differentiating keratinocytes would lead
to presence of the virus in all epidermal layers. In most
footpads, subcorneal keratinocytes were most intensively stained for virus particles, with less staining of
lower layers. Viral infection of basal keratinocytes may
have occurred during the viremia that followed the
experimental inoculation. In this case, basal keratinocytes would have been exposed to a heavy viral load
and the keratinocytes in the subcorneal layer could
represent those basal cells after migration. The time
span of keratinocyte differentiation and migration in
canine footpad epidermis has not been published. In
humans a duration of about 2550 days is described for
the time a basal keratinocyte is shedded as a squame
with the shorter time being observed in areas exposed
to frictional forces like hand palms.24 Assuming that
keratinocyte maturation in the footpad of the dog
occurs in a comparable time frame, the presence of
most intense CDV staining in subcorneal keratinocytes
would seem to indicate such comigration of the virus.
Alternatively, the specific cellular conditions in a keratinocyte at this particular stage of maturation may be
such as to support viral replication more successfully
than keratinocytes in another phase of development.
However, the presence of individual keratinocytes in
lower epidermal layers including basal cells, which
were strongly positive for nucleoprotein, implies that
viral replication may benefit from these conditions but
does not depend on them.
It was not possible to correlate the distribution of
CDV in the epidermis with duration of infection. The
presence of CDV particles in basal footpad keratinocytes exemplifies this point: dogs 4 and 5 were infected
4 days longer than dog no. 2 and 8 days longer than
dog no. 1, which had no viral nucleoprotein in the basal
layer. This finding would support the notion that the
distribution of CDV within the epidermis is not only a
function of time.
CDV antigen and mRNA was demonstrated in footpads of group 1 dogs with severe distemper and CNS
lesions. The association between digital hyperkeratosis
and encephalitis has already been observed by the first
authors to describe hard pad disease and others.22,23
Infection of the CNS and footpad epidermis probably require similar conditions regarding virus load,
specific viral virulence factors or disposition of the
host. Potentially, footpad keratinocytes, like the CNS,
are infected during the secondary viraemia and not
during the initial viraemia following inoculation. Such
a scenario would explain the contemporaneous occurrence of the virus in the CNS and footpad epidermis,
and lack of virus in the footpad epidermis of distemper
dogs without CNS lesions.

165

A span of 210 weeks is given in the literature for the

development of CDV-induced digital hyperkeratosis.25
The absence of this change in the footpad epidermis
in this study was not expected because the duration
between inoculation and death fell within this time
frame. The cause remains unclear; however, the most
likely reason may still be time, as the dogs with CDV in
the footpad epidermis were euthanized about 3 to 4
weeks after inoculation and may not have had enough
time to develop footpad hyperkeratosis. Absence of
hyperkeratosis of footpads and nasal planum was also
noted in the majority of cases in a study of naturally
infected distemper dogs, but as the presence of CDV in
the footpads of those dogs is not known this could also
be because there was no virus in the footpads.26
In contrast to descriptions in the literature, in this
study CDV infection of the footpad epidermis rarely
caused cellular degeneration, indicating that this
persistent infection was not cytocidal. Comparable noncytocidal infection by CDV has been observed in the
brain, and restricted translation of viral proteins is
discussed as a presumptive cause of persistent infection.2730 However, in this study nucleoprotein mRNA
colocalized with nucleoprotein antigen and the distribution of staining intensity was similar. This finding
disputes the idea that the lack of cytolytic changes may
be due to restricted protein translation.
Overall, there were not many pathological changes
on routine histopathological evaluation. Immunohistochemistry allowed detection of higher numbers
of CD3-positive T cells and of apoptotic cells in the
epidermis of group 1 dogs. Infiltration of T cells is
observed in encephalitic brains of dogs with distemper
and therefore a T-cell response in the epidermis as suggested by these findings was not surprising.31,32 Overall, however, the number and distribution of T cells
appeared very limited when compared to the many and
widespread keratinocytes positive for CDV nucleoprotein. These findings parallel observations from
footpads of naturally infected dogs, indicating that
inflammation may not be a prominent feature of CDV
infection of footpad epithelium.12
Apoptosis as a consequence of CDV infection has
been observed in vitro using the Onderstepoort vaccine
strain.33 In contrast, apoptosis is not a significant feature in in vitro studies using the same virulent strain as
the one in this study (A75/17), and apoptosis could not
be observed in brains of naturally infected distemper
dogs.15 Low numbers of apoptotic cells in canine footpad epidermis, as observed in dogs of all groups, are
reported in the literature.34 Perhaps apoptosis in canine
footpad epidermis is a more easily induced response to
cellular injury compared to the CNS. One could speculate that the increased numbers of apoptotic cells are
secondary to CDV infection; however, they are more
likely to be related to the higher numbers of T cells that
were present in these samples.
In summary, after experimental infection, CDV
antigen and mRNA were found in keratinocytes at all
levels of canine footpad epidermis, indicating persistent

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A Grne et al.

infection. Hyperkeratosis was not a feature and neither

degeneration nor inflammation was prominent.

ACKN OWLEDGE ME NT S
The authors thank the Berna Biotech AG, Berne, Switzerland, for financial support, Ines Schmid and Marianne
Wyder for excellent technical assistance, and especially
Claudia von Tscharner for critical review of this
manuscript.

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Canine distemper virus in canine footpads

167

Rsum Linfection de lpiderme des coussinets par le virus de la maladie de Carr (CDV) peut apparatre dans
lvolution naturelle de cette maladie. Cette tude sest intresse dcrire lexamen histopathologique des coussinets de 19 chiens ayant subi une inoculation exprimentale de la souche A75/17 de maladie de Carrr et de 2
chiens non exposs. Dans tous les cas, la prsence dantigne viral tait recherche, ainsi que lARNm des nucloprotines et le nombre de cellules inflammatoires et apoptotiques. Les chiens ont t diviss en quatre groupes
en fonction de leur statut aprs inoculation et de lexamen autopsique: chiens inoculs prsentant une maladie
de Carr svre (groupe 1, n = 7), chiens inoculs prsentant une maladie de Carr modre (groupe 2, n = 4),
chiens inoculs sans maladie de Carr (groupe 3, n = 8), and chiens non inoculs (groupe 4, n = 2). Les coussinets
des chiens de tous les groupes avaient un piderme pais de taille quivalente. Des inclusions virales osinophiliques et des cellules syncitiales taient prsentes dans lpiderme des coussinets dun chien du groupe 1.
Les coussinets des chiens du groupe 1 contenaient des antignes viraux et de lARNm avec un marquage important en position sous-corne. En outre, chez ces chiens, des structures du derme, notamment les glandes eccrines
et les parois vasculaires, taient positives pour le virus. Aucun antigne viral ou ARNm na t observ pour tous
les autres chiens. Les chiens du groupe 1 prsentaient plus de cellules CD3+ et de cellules apoptotiques dans leur
piderme que les autres groupes. Ces observations dmontrent quen cas dinfectioni exprimentale, les antignes
viraux et lARNm sont localiss dans toutes les couches de lpiderme des coussinets. La pauvret de la raction
inflammatoire associe, avec absence de dgnrescence des kratinocytes montre une infection persistente non
cytolytique des kratinocytes des coussinets par le virus de Carr.
Resumen En la infeccin natural por el virus del moquillo canino (VMC) puede producirse infeccin de la epidermis del almohadilla. Las almohadillas de 19 perros inoculados experimentalmente con la cepa virulenta de
moquillo A75/17 y de dos perros no expuestos fueron evaluadas histopatolgicamente y se valor la presencia
de antgeno vrico y mRNA de nucleoproteina, as como el nmero de clulas inflamatorias y apoptticas. Los
perros fueron divididos en cuatro grupos basados en funcin de si haban recibido inoculacin y del examen postmortem: perros inoculados con moquillo grave (grupo 1, n = 7), perros inoculados con moquillo leve (grupo 2,
n = 4), perros inoculados sin moquillo (grupo 3, n = 8), y perros no-inoculados (grupo 4, n = 2). Las almohadillas
de todos los perros mostraban una epidermis comparativamente gruesa. Se hallaron inclusiones vricas eosinoflicas y clulas sincitiales en la epidermis de la almohadilla de un perro del grupo 1. Las almohadillas de los perros
del grupo 1 contenan antgeno vrico y mRNA en la epidemis con mayor intensidad de tincin a nivel subcorneal.
Adems, en las almohadillas de estos perros, las estructuras drmicas incluyendo las glndulas ecrinas y las paredes vasculares fueron positivas a las partculas vricas. No se encontr VMC ni mRNA en la epidemis de la almohadilla ni en la dermis de ningn otro perro. Los perros del grupo 1 tenan ms clulas CD3-positivas y clulas
apoptticas en la capa basal de la epidermis que los de los dems grupos. Estos hallazgos demuestran que en la
infeccin experimental, el antgeno y mRNA del VMC se encontraban co-localizados en todas las capas de la
epidermis canina de las almohadillas infectadas. La escasez de reacciones histopatolgicas claras con la ausencia
de degeneracin de queratinocitos indica una infeccin persistente no-citocida de los queratinocitos de la almohadilla por parte del VMC.
Zusammenfassung Bei der natrlichen Infektion mit caninem Staupevirus (CSV) kann es bei Hunden zu einer
Infektion der Epidermis der Fusohlen kommen. Die Fusohlen von 19 Hunden, die experimentell mit dem virulenten Staupe-Stamm A75/17 inokuliert wurden und von zwei nicht exponierten Hunden wurden histopathologisch untersucht und in Hinblick sowohl auf das Vorhandensein von viralem Antigen und NukleoproteinmRNA als auch auf die Anzahl von inflammatorischen und apoptotischen Zellen hin berprft. Aufgrund des
Inokulationsstatus und der post-mortem Untersuchung wurden die Hunde in 4 Gruppen eingeteilt: inokulierte
Hunde mit starker Staupe (Gruppe 1, n = 7), inokulierte Hunde mit milder Staupe (Gruppe 2, n = 4), inokulierte
Hunde ohne Staupe (Gruppe 3, n = 8) und nicht inokulierte Hund (Gruppe 4, n = 2). Die Fusohlen der Hunde
aller Gruppen hatten vergleichbar dicke Epidermis. Bei einem Hund der Gruppe 1 waren eosinophile virale Einschlusskrperchen und Synzytialzellen in der Epidermis der Fusohlen vorhanden. Die Fusohlen der Hunde
von Gruppe 1 enthielten virales Antigen und mRNA in der Epidermis mit der strksten Anfrbung in einer subkornealen Lokalisation. Bei den Fusohlen dieser Hunde waren dermale Strukturen einschlielich der eccrinen
Drsen und der Gefwnde positiv fr Viruspartikel. Bei keinem der anderen Hunde war CSV-Antigen oder
mRNA in der Epidermis oder Dermis der Fusohlen vorhanden. Die Hunde der Gruppe 1 hatten im Vergleich
zu den anderen Gruppen mehr CD3-positive und apoptotische Zellen in der Basalschicht der Epidermis. Diese
Ergebnisse zeigen, dass bei experimenteller Infektion CSV-Antigen und mRNA zusammen in allen Schichten der
infizierten caninen Epidermis der Fusohlen zu finden sind. Die Seltenheit offensichtlicher pathologischer Reaktionen zusammen mit dem Fehlen von Keratinozytendegeneration weist auf eine nicht-zytozide persistierende
Infektion der Keratinozyten der Fusohlen durch CSV hin.

2004 European Society of Veterinary Dermatology, Veterinary Dermatology, 15, 159167