Veterinary Dermatology 2004, 15, 207 217

Hereditary equine regional dermal asthenia (hyperelastosis cutis)

in 50 horses: clinical, histological, immunohistological and
ultrastructural findings

Blackwell Publishing, Ltd.

STEPHEN D. WHITE*, VERENA K. AFFOLTER, DANIKA L. BANNASCH,

PATRICIA C. SCHULTHEISS, DWAYNE W. HAMAR, PHILLIP L. CHAPMAN**,
DIANE NAYDAN, SHARON J. SPIER*, ROD A. W. ROSYCHUK, CHRISTINE REES,
GREGG O. VENEKLASEN, ALONDRA MARTIN, DIANE BEVIER,
HILARY A. JACKSON, SONYA BETTENAY, JENNIFER MATOUSEK***,
KAREN L. CAMPBELL*** and PETER J. IHRKE*
*Department of Medicine and Epidemiology,
Department of Pathology, Microbiology and Immunology,
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California,
Davis, California 95616, USA
Veterinary Diagnostic Laboratory,
Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences,
**Department of Statistics, Colorado State University, Fort Collins, Colorado, 80526, USA
Department of Small Animal Medicine & Surgery, College of Veterinary Medicine, Texas A & M University,
College Station, Texas 778434474, USA
Timber Creek Veterinary Hospital, Route 2, Box 366, Canyon, Texas 79015, USA
Department of Clinical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette,
Indiana 47907, USA
Department of Companion Animal Practice, North Carolina State University, College of Veterinary
Medicine, 4700 Hillsborough Street Raleigh, North Carolina, 27606, USA
***Department of Veterinary Clinical Medicine, University of Illinois Urbana, Illinois 61802, USA
(Received 23 August 2003; accepted 15 March 2004)

Abstract Data on fifty horses with hereditary equine regional dermal asthenia (HERDA; hyperelastosis cutis)
were collected on clinical, histopathological, ultrastructural and immunohistological findings. All horses were
Quarter horses or of Quarter horse ancestry. Pedigree evaluation strongly supported an autosomal recessive mode
of inheritance. The most common lesions were seromas/haematomas, open wounds, sloughing skin, and loose,
easily tented skin that did not return to its initial position. Definitive diagnosis could not be made via histopathology, although the presence of tightly grouped thin and shortened collagen fibres arranged in clusters in the
deep dermis was suggestive of the disease. Trichrome, acid orcein-Giemsa and immunohistochemical stains for
collagens I and III showed no consistent abnormalities compared to control horses; an increase in elastic fibres
was not a consistent finding. Electron microscopy showed no abnormalities in the periodicity of the collagen
bundles; neither orientation nor variation of cross-section diameter of the collagen fibrils differentiated control
from affected horses. The diagnosis of HERDA relies on clinical presentation, but may be supported by suggestive (although not pathognomonic) histopathological lesions.
Keywords: collagen, HERDA, hereditary equine regional dermal asthenia, horses, hyperelastosis cutis, skin.

IN TRO D U CT I ON
A skin disease affecting young Quarter horses has been
reported since 1978, in which the skin is described
as hyperelastic, fragile and thin, with slow-healing
wounds characterized by atrophic scars.14 Lesions
may be single or multiple, and are most commonly seen

Correspondence: S. D. White, Department of Medicine and

Epidemiology, School of Veterinary Medicine, University of California,
Davis, California 95616, USA. E-mail: sdwhite@ucdavis.edu
2004 European Society of Veterinary Dermatology

on the dorsum, although they may also occur on the

legs.15 Frequently, the skin in nonscarred, affected
areas is described as depressed, with an ability to be
grasped and stretched beyond that of normal equine
skin.1,4,5 A clinically similar disease has been reported
in a cross-bred Arabian mare, a thoroughbred gelding,
a Hanoverian foal and a Hafflinger horse.69
Previous articles have dealt with one or two affected
horses. The purpose of this combined retrospective/
prospective study is to report the clinical, histopathological, ultrastructural and immunohistochemical
findings of 50 horses affected with this disease.
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SD White et al.

M ATERIALS AND ME T HODS

Selection of cases
Horses were included in the study if they had clinical
features as defined by Stannard: young horses with welldemarcated areas of fragile and loose skin, associated
with poorly healing ulcers and/or haematomas/seromas.5
Retrospective cases were selected from records kept by
the late Dr Tony Stannard at the University of California
at Davis (UCD) and maintained by one of the authors
(SJS). Prospective cases were either horses examined by
at least one of the authors, or consecutive biopsy specimens sent to the authors; the referring veterinarians
were then contacted to supply clinical information.

ated at 0, 25, 50, 75, 90 and 100%. All available

histology samples were evaluated; however, in
order to eliminate the possibility of different tinctorial qualities based on different staining runs,
six samples were re-stained in the same run.
These six samples were chosen from four affected
and two healthy Quarter horses; the horses were
among those that both pathologists agreed were
either certain to be affected or unaffected (see E
below). These six samples were blinded by the
first author and the percentage of normal staining evaluated by one of the pathologists (VKA);
D. Number of elastic fibres: 1 = normal, 2 = slightly
more than normal; 3 = many more than normal;

Data
The following information was collected: age, age of
onset, breed, sex, type of lesions, distribution of lesions
and pedigrees.

Histopathology
Biopsy specimens (either 6 mm punch or incisional
wedge) were taken from the lesions, or if the lesion was an
ulcer, from the immediately adjacent tissue. When possible, biopsies were also taken from the following areas,
whether lesional or not: side of neck (approximately
where the reins would contact the skin), withers, croup,
saddle area, elbow, knee and pastern (front and rear).
Biopsy tissues were submitted in 10% buffered formalin, routinely processed to paraffin and stained with
haematoxylin and eosin (H&E), Masson trichrome
(MT) and acid orcein-Giemsa (AOG). Control skin
specimens were obtained from eight healthy Quarter
horses without evidence of skin disease. Three of these
healthy Quarter horses were < 3 years of age and were
related (siblings or parents) to affected horses. The
other healthy Quarter horses were unrelated to the
affected horses and were > 3 years of age. Biopsy
tissues were also obtained from three healthy nonQuarter horses (two Thoroughbreds and one Haflinger).
One of the Thoroughbreds was < 1 year of age and the
other two horses were > 5 years of age. Slides of the
biopsy tissues were blinded by the first author and subsequently examined by two of the authors, diplomates
of either the European College of Veterinary Pathologists (VKA) or the American College of Veterinary
Pathologists (PCS). Biopsies were evaluated subjectively on an ordinal scale for the following attributes:
A. Collagen fibre arrangement: 1 = fibres close together
(normal); 2 = single fibres separated from each
other (by oedema or fibrosis); 3 = tightly grouped
fibres arranged in clusters, separated from other
clusters;
B. Collagen fibre thickness: 1 = very thin; 2 = slightly
thin; 3 = normal;
C. Percentage of normal (blue) staining of collagen
fibres with Massons trichrome. These were estim-

E. Overall assessment: 0 = unlikely to be affected;

1 = suggestive of the disease; 2 = very suggestive
of the disease; 3 = certain of being affected.
Certainty was based on the presence of collagen
abnormalities in the deep dermis, most specifically thin fibres arranged in clusters instead of being
elongated, and the presence of clear spaces separating the clusters. As previously described, the deep
dermis was defined as extending from the base of
the anagen hairs to the cutaneous muscle.3,10
In addition, the biopsy specimens were evaluated for
the presence or absence of inflammation, trauma (tearing), and fibrosis. The recent description of zonal
dermal separation (ZDS) in an affected Quarter horse
prompted evaluation of our samples for this feature as
well.4 ZDS was defined as a horizontal linear zone of
loose collagen bundles located through the middle of
the deep dermis, with variable separation of collagen
bundles and loss of dermal density.4 The authors of
that report hypothesized that only deep biopsies would
be able to discern the zonal dermal separation; therefore,
we also noted the number of horses with biopsies showing
panniculus tissue at the deep margin of the dermis.

Statistical methods
A preliminary repeated measures analysis indicated
that the comparison between group mean responses
depended on pathologist; therefore, variables were analysed
separately for each pathologist. Variables measured on
the 03 or 13 scales (collagen fibre arrangement, collagen fibre thickness, percentage of normal staining of
collagen fibres with MT, number of elastic fibres and
overall assessment) were compared by group using ttests computed by SAS Proc TTEST (SAS Institute
Inc., Cary, NC, USA). The group variances were pooled
when the Folded F-test for equality of variances was
not significant. When the Folded F-test was significant,
the variances were not pooled, and Satterthwaite degree
of freedom estimates were used. Variables measured
on a 01 scale (presence or absence of inflammation,
tear or trauma and fibrosis) were compared by group
using Fishers Exact test. For all tests, P < 0.05 indicated significance. Statistical computations with control

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HERDA in horses
variables were performed both with and without the
findings from the three non-Quarter horses.

Immunohistochemistry
Based on reports of similar diseases in humans, we
elected to stain sections for collagen I, III and V.1114
The biopsy sections tested were chosen from six horses:
five affected and one healthy control. These particular
horses were among those that both pathologists agreed
were either certain to be affected or unaffected (see E
above).
Each selected biopsy tissue was fixed in neutral buffered 10% formalin and routinely processed to paraffin.
Serial sections, 4 microns in width, were cut and
mounted on plus coated slides. Immunohistochemical staining was performed using a standard streptavidin biotin-HRP technique as previously described15
with the following modifications for paraffin: sections
were deparaffinized in two 10-min changes of xylene
followed by gradual rehydration through alcohol to
phosphate buffered saline, pH 7.2. Commercial antibody kits were used. According to the manufacturers
data sheets, the antibody against collagen I was specific
to human collagen I, the antibody against collagen III
was specific to human and bovine collagen III, and
the antibody against collagen V reacted with human,
rabbit and rat collagen V.
Tissue antigens in the horse biopsy tissues to be
reacted with rabbit polyclonal collagen I (NovoCastra,
1 : 200, Vector Laboratories [distributor], Burlingame,
CA, USA) were heat retrieved in citrate buffer pH 6
(Dako Corporation, Carpinteria, CA, USA) for
30 min using steam, followed by cooling for 20 min.
Tissue antigens in the horse biopsy tissues to be reacted
with goat polyclonal collagen III (1 : 200, Chemicon
International, Temecula, CA, USA) and mouse monoclonal collagen V (1 : 250, Chemicon International)
were heat retrieved in Glyca pH 3 (Biogenex Laboratories, San Ramon, CA, USA) for 20 min in a procedure
similar to that for collagen I. The secondary, biotinylated reagent and the tertiary, streptavidin reagent
were provided as a predilute kit form (Biocare Medical,
Walnut Creek, CA, USA) and applied for 10 min each
at room temperature.
Negative controls were prepared by omitting the
primary antibody and substituting the appropriate
mouse, goat or rabbit IgG correlate. The immunostained sections were blinded by the first author, and
evaluated by one of the pathologists (VKA). These
were subjectively graded as well stained or not. Statistical evaluation was not performed, because of the use
of only one healthy control horse.

Ultrastructure
Biopsy tissues were collected in full-strength modified
Karnovskys fixative, then transferred to half-strength
Karnovskys fixative before postfixation in 2% osmium
tetroxide reduced with 2.5% potassium ferrocyanide.16,17 Following fixation, tissue was washed in 0.2
sodium cacodylate, and dehydrated through a graded

209

ethanol series before infiltration and embedment in

Spurrs epoxy resin for 48 h.18 Thick sections were cut,
mounted on glass slides and stained with Toluidine
blue O and examined by light microscopy. Thin
sections were cut, mounted on 150 mesh copper grids,
stained with 6% methanolic uranyl acetate and
Reynolds lead citrate, and examined in a Zeiss 10C
transmission electron microscope (Carl Zeiss electron
microscopes, Thornwood, NY, USA) at 60 kv accelerating voltage.19 Biopsy tissue from one of the healthy
adult Quarter horses noted above was used as a control.
Electron micrographs were blinded by the first
author and subsequently subjectively evaluated by the
two pathologists using the following parameters:
A. Orientation of the collagen fibrils: random = 1 or
parallel = 0;
B. Periodicity of the collagen bundles: variable = 1
or uniform = 0;
C. Cross-section diameter of
variable = 1 or uniform = 0.

collagen fibrils:

The scores from the two pathologists were combined. Thus, a severely affected horse should have a
score of 6 (3 possible from each pathologist) while the
healthy control should have a score of 0. Statistical
evaluation was not performed, because of the use of
only one healthy control horse.

R E SU LT S
Clinical data
Fifty horses were included in the study: eight were
retrospective cases, and 42 were prospective cases.
Thirty-three horses were examined by at least one of
the authors, and 17 were biopsy specimens sent in
by referring veterinarians; six of these samples were
accompanied by photographs of the affected horses.
Forty-three Quarter horses, one Quarter horse
cross-bred, four American Paint horses, and two
Appaloosa horses were included in the study. Twentyfour horses were mares, 15 were stallions, and 11 were
geldings. The age at presentation ranged from 0.6 to
6 years, with a mean of 2.4 and a median of 2. The age
of onset was known in only 29 horses, with a range of
birth to 4 years, a mean of 1.3 and a median of 1.
Full pedigrees were available for 42 horses, incomplete (lacking several ancestors) pedigrees were available for two American Paint horses, the sires pedigree
only was available for two Quarter horses, and no pedigree was available for three Quarter horses and the
Quarter horse cross. All 42 horses with full pedigrees
(38 Quarter horses, two American Paint horses and
the two Appaloosa horses) had one common ancestor
(termed here CA-1, a stallion) that appeared in both
the sires and dams pedigrees. Forty of these horses
also had another common ancestor (CA-2, also a

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SD White et al.

Figure 1. Eleven-month-old Quarter horse mare with HERDA.

Note easily wrinkled affected dorsal thoracic skin.

stallion) that appeared on both sides of their lineage;

CA-1 and CA-2 were not related. Analysis of the
pedigrees of the Quarter horses strongly supported an
autosomal recessive trait; this has been reported in
detail elsewhere.20
The two Quarter horses with only the sires pedigree
had both CA-1 and CA-2 in their lineage. Of the two
American Paint horses with incomplete pedigrees, one
had CA-1 and CA-2 on the sires side, and the other
had both on the dams side.
Owners presenting complaints were available in
38 horses. (The other 12 horses were included in this
study on the basis of physical examination findings as
reported by the veterinarian.) These included swellings
(seromas and or haematomas; often described as
blisters) (19 horses), scars (12), loose skin (11), open
wounds or sloughing skin (10), easily damaged skin
upon trauma (9), poor healing of wounds (7), and
white hairs at areas of hair re-growth (2). Seven owners
related the onset of clinical signs to the beginning of
saddle training. None of the geldings owners reported
a problem with healing from the castration surgery;
one horse had a problem with the biopsy site not healing. One horse had a twin that was unaffected.
The lesions most commonly noted on physical examination were swellings (seromas and or haematomas)
(24 horses), open wounds or sloughing skin (14), loose
easily tented skin which did not return to its initial
position (18), scars (16), and white hairs at areas of hair
re-growth (6). The lesions were seen over the dorsum
(withers to croup) (38 horses), gluteal region (13), neck
(8), lateral thorax or abdomen (6), distal legs (5), face (5),
tail-head (3) and/or coronary band (2) (Figs 14).

Figure 2. Two-year-old Quarter horse mare with HERDA. Note

resolving seroma with loose skin. White hair occurred subsequent to
seroma.

Figure 3. Three-year-old Quarter horse gelding with HERDA. Note

scarring and ulcers from sloughed skin.

Histopathology
Histopathology was performed on samples from 48
horses.
Forty-six of these samples were available for additional staining for this study. Both pathologists subjectively thought that the most consistent and suggestive
finding was the presence of collagen abnormalities

Figure 4. Same horse as Fig. 2. Note nonhealing wound on distal leg.

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HERDA in horses

Figure 5. Photomicrograph of the skin from a 3-year-old Quarter

horse gelding with HERDA. Note the presence of thin and
shortened deep dermal collagen fibres arranged in clusters, separated
by clear spaces. (H&E, bar = 500 m.)

Figure 6. Photomicrograph of the skin from the same horse as

Fig. 5, higher magnification, deep dermis. (H&E, bar = 100 m.)

in the deep dermis. The most consistent change in

affected tissues was the presence of thin and shortened
deep dermal collagen fibres arranged in clusters, separated by clear spaces (Figs 5 and 6); control tissues
were characterized by having thick and long, evenly
distributed collagen fibres. The clear spaces separating
the small clusters of fibres in affected horses appeared
either empty or contained a few thin fibres. Fibrosis
and/or granulation tissue in the mid to deep dermis was

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Figure 7. Photomicrograph of the skin from a 2-year-old Quarter

horse mare with HERDA. Note normal blue staining of the thin and
shortened deep dermal collagen fibres. (Massons trichrome,
bar = 300 m.)

Figure 8. Photomicrograph of the skin from a 3-year-old Appaloosa

stallion with HERDA. Note dermal zonal separation. (H&E,
bar = 750 m.)

noted in 21 affected horses, two of the healthy Quarter

horses (one of these was from the three related but
unaffected horses), and one of the healthy non-Quarter
horses. This most probably indicated a previous separation of tissues with subsequent scarring. The epidermis,
hair follicles and adnexa were normal in all horses.
With regards to the MT stain, in 44 of the affected
horses more than 75% of the collagen fibres stained

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SD White et al.

blue (normal), without red cores (in 29 of the affected

horses 90100% of the fibres stained blue) (Fig. 7). The
six samples evaluated with MT stain on the same run
showed the control horses having the lowest percentages (2030%) of normal staining fibres.
The following statistical analyses are based on using
only the healthy Quarter Horses for the controls. Differences in statistical significance that occurred if the
non-Quarter horses were included are discussed in the
next paragraph. Of the two pathologists, VKA found
statistically significant differences between the two groups
(affected vs. control) for arrangement of collagen fibres
(P = 0.0014), percentage of fibres staining blue (normal)
with MT stain (P = 0.024), overall assessment (i.e. certainty of having the disease) (P = 0.0044), and inflammation (P = 0.015). The other pathologist (PCS) found
a statistically significant difference between the affected
horses and the healthy controls with regards to thickness of the collagen fibres (P = 0.046) and inflammation
(P = 0.0001), although her differences for arrangement
of collagen fibres (P = 0.067), and overall assessment
(P = 0.28), had estimated group differences in the same
directions (trends) similar to those of VKA. The percentage of fibres staining blue (normal) with MT stain
according to PCS was not significant (P = 0.27). Neither pathologist found a statistically significant difference between affected and control horses with regards
to the number of elastic fibres (P = 0.21 [VKA], P = 0.49
[PCS]), evidence of trauma (P = 0.88 [VKA], P = 0.93
[PCS]), or the presence of fibrosis (P = 0.106 [VKA],
P = 0.98 [PCS]).
Inclusion of the three healthy non-Quarter horses in
the controls changed the significance (or not) of only
three of the above statistics. Interestingly, it changed
both pathologists findings with regards to the thickness of the collagen fibres, VKA changing to significance
(P = 0.0072) and PCS to nonsignificance (P = 0.14).
With regards to the percentage of fibres staining blue
(normal) with MT stain, VKA changed to nonsignificance (P = 0.066).
Comparing the two pathologists findings, there was
no difference in the statistical significance between the
pathologists in their evaluation of horses with regards
to arrangement or thickness of collagen fibres, inflammation and trauma. While one pathologist may have
found statistically significant differences between the
two groups of horses with these parameters and the other
pathologist did not, their evaluations were fairly consistent (i.e. one just made the statistically significant
cut-off, and the other did not). There was a difference
in the statistical significance between the pathologists
in their evaluation of overall assessment, percentage of
fibres staining blue and the number of elastic fibres.
Ten of the affected horses also had biopsies submitted from healthy-appearing skin, which were admixed
into the blinded slides for those horses. VKA assigned
an overall assessment score of either 2 or 3 (2 = very
suggestive of the disease; 3 = certain of being affected)
to nine of these horses, while PCS assigned an overall
assessment score of either 2 or 3 to six of these horses.

Figure 9. Electron micrograph of the skin from a 3-year-old Quarter

horse gelding with HERDA. Note random orientation of collagen
fibrils. (Uranylacetate, bar = 50 m.)

Two horses were necropsied; internal organ lesions

were not noted. Only one example of zonal dermal separation, similar to that previously described, was noted
(Fig. 8).4 Eighteen of the 50 affected horses had deep
biopsies.

Immunohistochemistry
The six cases (five affected and one healthy control)
evaluated for immunostaining all were judged to stain
well for collagen I and III, with no difference discerned
between the healthy control and the affected horses.
We were unable to develop a staining technique that
consistently stained for collagen V in either the healthy
control or affected horses.

Ultrastructure
Electron microscopy (EM) was performed on samples
from 13 affected horses, plus one healthy control horse.
All horses were judged to have normal periodicity of
the collagen fibres. Eight of the affected horses and the
control horse showed randomness in the orientation of
the collagen fibrils (Figs 9 and 10); 12 of the affected
horses and the control horse showed variability of the
cross-sections of the collagen fibrils. No horse had a
score of 0 (unaffected) or 6 (severely affected). Of the
affected horses, the mean score was 2.8, with a median
of 2. In contrast, the healthy control had a score of 4.

D ISC U S S IO N
The name hyperelastosis cutis probably was meant to
apply to the loose, easily tented skin of these horses,
as reported in one of the early publications, which
described a case of cutis hyperelastica.6 However,

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HERDA in horses

Figure 10. Electron micrograph of the skin from a healthy 5 year-old

Quarter horse mare. Note random orientation of collagen fibrils.
(Uranyl acetate, bar = 50 m.)

elastosis is defined as degeneration of elastic tissues or

fibres21 and hyperelastosis as excessive deposits of
elastic fibres.22 Neither of these elastic pathologies was
seen in the horses in this study, supporting a similar
observation in a previous report.3 We therefore suggest
the name Hereditary Equine Regional (referring to the
propensity of the lesions of this disease to affect only
certain areas of the body) Dermal Asthenia (reflecting
the fact that the defect is in the dermis loss of normal
strength),21 with the acronym HERDA.
The signalment data are similar to previous reports,
with the disease occurring equally in males and females
and in Quarter horses or horses with Quarter horse
ancestry.5,8 The median age of onset and age of examination are also consistent with a disease of young
horses, although horses may be bought and sold such
that the owner who sought veterinary care for the disease may not have had the knowledge of when the
disease actually was first noted.
We feel confident that of the 33 horses we have examined, and the six horses examined by referring veterinarians that were accompanied by photographs, all
had HERDA. The physical examination and historical
criteria of well-demarcated areas of fragile and loose skin,
associated with poorly healing ulcers and/or haematomas/seromas were felt to be exclusive of other diseases,
thus eliminating in the other 11 horses the possibility
of inclusion of a healthy horse with one or more scars.
The autosomal mode of inheritance of this disease
among Quarter horses has been theorized2,4 and
reported.20 Interestingly, one of the affected horses twin
was unaffected. While twins in horses are uncommon,
they are almost always nonidentical, and are the result

213

of two ovulations: two oocytes that become fertilized

during the same cycle.23 Thus, only one twin inherited
the presumptive defective gene from both parents.
Clinical signs of the horses in this report were generally comparable with those in the literature.15 A striking
clinical finding in this disease is the relatively limited
areas of the body (usually the dorsal trunk) affected.
The reason for this restriction is not entirely clear.
While observation by several owners relating the onset
of clinical signs to the beginning of saddle training
would seem to imply that lesions arise in areas subject
to friction or trauma, similar lesions were not noted on
the ventrum, where the cinch would contact the skin,
nor as a sequela to castration in the geldings. We were
unable to find studies of wound healing in horses skin
that compared the dorsum with other areas of the
body. However, as 10% of the affected horses in this
report had distal leg lesions, the findings of a recent
study on wound healing in horses that compared skin
wound healing on the distal leg to the pectoral thorax
may have some relevance. 24 In that study there was
a greater accumulation of collagen in the distal leg
wounds than in those on the pectoral thorax. This was
postulated to be due to an imbalance of collagen synthesis and degradation.24 If there is a defect in the collagen of the skin in the HERDA horses, perhaps there
is a greater accumulation of this defective collagen in
the lower leg which predisposes to lesions occurring
there, at least compared to the pectoral area, where
HERDA lesions seldom occur. Alternatively, if there
is increased collagen synthesis in the distal limb of
HERDA horses, perhaps this somehow compensates
for the disease, which may explain the paucity of leg
lesions compared to dorsal lesions in affected horses.
The admixing of biopsies from healthy-appearing
skin with the biopsies from clinically affected skin did
not seem to affect the overall assessment of the majority of the 10 horses thus studied. This would suggest
that histopathological changes may be present in the
absence of obvious clinical features. This may support
a previous report, which stated that the areas of the
affected skin may be more extensive on necropsy than
noted clinically.5
While poor healing of wounds was reported by some
owners, it is difficult to assess whether there is a true
defect in the healing process, or whether the presence
of the dermal defect merely leads to new wounds in the
same area upon new trauma. Previous reports differ,
with some noting normal healing2 and others delayed.35
Again, the fact that none of the geldings had a problem
healing from the castration surgery would at least
imply that any problem of healing is, like the lesions
themselves, restricted to certain areas of the skin.
Our histopathological findings in this study are similar to previous reports, which describe thinning, fragmentation, and disorientation of the collagen fibres
in the mid to deep dermis, often separated by empty
spaces.1,3,5,7,9 These empty spaces may have been the
result of a protein-poor oedema, which would be
removed by the alcohol treatment used to prepare the

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SD White et al.

paraffin sections. Alternatively, these spaces could represent the result of defective adhesiveness of the fibre
bundles and subsequent separation due to handling,
i.e. a useable artifact as defined by Stannard.25 The
statistical significance of the arrangement and thickness of the collagen tissue between affected and nonaffected horses varied according to the pathologist and
whether the healthy non-Quarter horses were included
in the controls. However, the lack of statistical significance between the evaluations of these parameters by
the two pathologists lends support to their subjective
observation that deep dermal collagen fibre abnormalities were the most consistent finding in HERDA horses.
We did note one example in an affected horse of
zonal dermal separation as previously reported in an
affected horse.4 The deep dermal position of the clusters of collagen fibres, plus evidence of fibrosis and/or
granulation tissue in the mid to deep dermis in 21
horses (as might be expected as a sequela to a dermal
separation of tissues) also could be supportive of ZDS
as a histological finding in HERDA horses. It also may
be a useable artefact: zonal dermal separation may
merely indicate that the defect in the dermis occurs at
this separation, and the separation was induced by performing the biopsy or by its processing. It should be
noted that fibrosis and/or granulation tissue in the mid
to deep dermis was seen in three of the healthy horses
(two of which were Quarter horses), supporting our
finding that there was no statistical difference between
these two groups in regards to the presence or absence
of trauma and fibrosis. To our knowledge, there are no
studies of healthy horses skin specifically to determine
histological evidence of past trauma; considering the
potential for trauma involved in putting on a saddle, riding,
etc., it seems likely that such minor injuries could occur.
Massons trichrome stain is used to distinguish collagen from other tissues, particularly muscle.26 Tissues
are first stained with a red stain (Biebrich scarlet-acid
fuschin), washed in an acid solution (which allows this
stain to diffuse out of the collagen fibres) then counterstained with either aniline blue or aniline green. One
text considers aniline blue the better stain when only
small amounts of collagen are present.26 Collagen thus
stains either blue or green, while abnormal collagen
fibres will have red cores.26 Two previous reports have
noted greater incidence of red cores in the skin of three
affected horses relative to three healthy horses.3,7 These
two reports used aniline green as the counterstain. In
contrast, a more recent report showed no staining
abnormalities in an affected horse.4 Both the recent
report and our laboratory methods used aniline blue.4
This technique has been shown to be adequate in distinguishing abnormal from normal collagen fibres in
inflammatory dermatoses in horses, and in cats in
contrasting the congenital disease cutaneous asthenia
from acquired skin fragility.27,28 Although one pathologists evaluation showed a slight statistical difference
between affected and control Quarter horses, the failure of most affected horses biopsies to show abnormal
red cores led us to question the diagnostic potential of

this stain. In the six cases stained on the same run, wherein the healthy horses had the lowest percentages of normalstaining collagen fibres, the denser packing of the collagen
fibres (as would be expected in normal skin) may have
reduced the rate of penetration of the counterstain,
thus leading to some red cores in healthy horses.29
Because of the original name of this disease, we were
interested in determining if elastic fibres were either
consistently more numerous than normal, or degenerated. The AOG stain may be used to distinguish elastic
fibres.30 We were unable to find any pathological
changes of the elastic fibres. Consistent with our findings of having no elastic changes associated with the
collagen changes, elastic fibres have been reported as
being independent of the collagen fibre bundle architecture in normal skin of the horse.31
It should be stressed that while the biopsies were
blinded, the pathologists did know that samples from
HERDA horses were being evaluated. We would caution that if no history were to be provided, the diagnosis may be missed, and that lack of histopathological
lesions does not negate the clinical diagnosis.
The EhlersDanlos Syndromes (EDS) in humans
are a heterogeneous group of heritable connective
tissue disorders characterized by skin hyperextensibility,
articular hypermobility, and tissue fragility. EDS is not
completely clinically analogous to the condition in
HERDA horses, as affected human skin does not show
body region variation, and none of the affected horses
in our study showed joint hypermobility or evidence of
internal organ collagen dysfunction. However, EDS is
caused by defects in fibrillar collagen formation.1214
These defects affect collagen types I, III or V.13,14 A
recent study identified a human patient with a defect in
collagen III in which immunohistochemistry stains
showed a loss of staining for this collagen, which was
confirmed by a decreased biochemical content of
collagen III when compared to normal controls.11 This
suggested that antibodies to distinct types of collagen
may be able to delineate the type of defective collagen in affected horses. Collagen I3,32,33 and collagen
III3 have been described in horses, and successful
immunostaining with antibody raised against human
epitopes for collagen types IV and VII has been
reported.34 However, our staining for collagen types I
and III showed no difference between affected horses
and a control horse. This supports a previous study of
two affected Quarter horses which found that collagen
I and III were present in similar proportions to control
horses.3 This suggests either a defect in a feature of collagen development, which does not affect the immunostaining properties of the collagen tissue, a defect in
collagen V (for which we were unable to achieve a consistent staining procedure), or a defect in a noncollagen
substance of the skin. The latter has recently been
reported in humans with tenascin-X deficiency (the
tenascins are extracellular-matrix proteins highly
expressed in connective tissue),14 and in humans and
cattle with a defect in a proteoglycan of the extracellular matrix.35,36

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HERDA in horses
In the biopsy specimens from 13 horses on which
electron microscopy was performed, randomness in
collagen fibril orientation and variability of collagen
fibril cross-sections was consistent with previous
reports of horses with this condition.3,4 These findings
are also consistent with ultra-structural studies of EDS
in humans37 and EDS-like diseases in other species.3842
Again, these diseases in other mammals, while causing skin tearing and fragility, affect skin over the entire
body, showing none of the regional variation seen in
the HERDA horses. Caution may further be warranted in over-interpretation of EM of affected horses,
as some of the same fibrillar changes were seen in the
control horse. Difficulties in interpretation of EM in EDS
in humans have been noted, with normal EM findings
of some human patients36 and damage (due to actinic
change, wound repair or specimen handling) to normal
tissue noted to cause some of the same types of fibril
abnormalities as the inherited collagen disorders.43,44
We do not suggest that EM is not an important aspect of
studying this disease, but rather that EM must be interpreted in conjunction with the history and clinical signs.
In conclusion, we describe a cohort of 50 Quarter
horses or horses of Quarter horse ancestry having an
autosomal recessive skin disease, whose diagnosis is
based on a constellation of clinical and histological
signs. Collagen dysfunction is suggested by the abnormalities seen in the deep dermis and perhaps the fibrillar changes seen by EM. Our findings suggest that the
defect does not involve the amount of collagen types I,
III or elastin. Further studies are currently underway
to delineate the genetic defect.

ACKN OWLEDGE ME NT S
This project was supported by the Center for Equine
Health with funds provided by the Oak Tree Racing
Association, the State of California paramutual fund,
and contributions by private donors. The project was
further supported by the George H. Muller Fund for
Research in Veterinary Dermatology.
The authors thank: (1) the following veterinarians
for providing information on affected horses: Kenton
Arnold, James Carmalt, Roger Cole, Sandy Curie,
Leonard Eldridge, Z. Franklin, Rohn Hendricks,
Nat Messer, Richard Mosier, William Moss, Roger
Rees, Royce Smathers, Amy K. Youngblood; (2) Mrs
Cynthia Hamar for sharing her knowledge of Quarter
horse ancestry and lineage; (3) Drs Linda Van Hoogmoed and Karen Terio for obtaining the skin biopsy
samples from healthy horses; and (4) Dr Kathy V.
Fieseler for technical support and Tracy Greenwalt for
technical and digital photography support.

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Rsum Cinquante chevaux prsentant une asthnie dermique hrditaire rgionale (HERDA; hyperelastosis
cutis) ont t tudis pour laspect clinique, histopathologique, ultrastructural et immunohistologique. Tous les
chevaux taient des Quarter horses ou avaient un Quarter horse dans leur ligne. Lvaluation des pedigree tait
en faveur dun mode autosomal rcessif de transmission. Les lsions les plus frquentes taient des sromes/
hmatomes, des plaies, une peau lche et la persistance dun pli de peau. Le diagnostic dfinitif na pas pu tre
ralis par lexamen histopathologique seul, bien que la prsence de fibres de collagne fines et de petite taille
dans le derme profond soient vocatrices de la maladie. evaluation strongly supported an autosomal recessive
mode of inheritance. Les colorations au trichrome, lacide orcein-Giemsa et des marquages immunohistochimiques pour les collagnes I et III nont pas montr danomalie en comparaison de chevaux sains. La prsence
dune augmentation des fibres lastiques ntait pas une observation systmatique. La microscopie lectronique
na pas montr danomalie des fibres de collagne ou de leur orientation par rapport des chevaux sains. Le
diagnostic dHERDA repose donc sur la prsentation clinique, et peut tre aid par les modifications
histopathologiques, vocatrices, mais pas pathognomoniques.
Resumen Se recopilaron los hallazgos clnicos, histopatolgicos, ultraestrucutrales e inmunohistolgicos de
cincuenta caballos con astenia drmica equina hereditaria regional (HERDA; hiperelastosis cutis). Todos
los caballos eran Quarter horse o tenan ancestros de esta raza. La evaluacin del pedigr indicaba claramente
una forma de heredabilidad autosmica recesiva. Las lesiones ms frecuentes fueron seromas/hematomas,
heridas abiertas, escarificacin de la piel, y piel suelta fcilmente desplazable y que no volva a su posicin inicial.
2004 European Society of Veterinary Dermatology, Veterinary Dermatology, 15, 207 217

HERDA in horses

217

El diagnstico definitivo no poda hacerse mediante histopatologa, aunque la presencia de fibras de colgeno
ntimamente agrupadas, delgadas y cortas, dispuestas en clusters en la dermis profunda era sugestiva de la
enfermedad. Las tinciones tricrmica, de cido orcena-Giemsa e inmunohistoqumicas para colgenos I y III
no mostraron anormalidades constantes comparando con los caballos control; no se observ de forma constante
un incremento en fibras elsticas. La microscopa electrnica no mostr anormalidades en la periodicidad de
los haces de colgeno, y tampoco la orientacin o la variacin del dimetro de las secciones cruzadas de las
fibras de colgeno permita diferenciar entre los caballos control y los afectados. El diagnstico de HERDA
se basa en la presentacin clnica, pero puede ser apoyado por lesiones histopatolgicas sugestivas (aunque no
patognomnicas).
Zusammenfassung Von fnfzig Pferden mit heriditrer equiner regionaler dermaler Asthenie (HERDA; Hyperelastosis cutis) wurden Daten bezglich klinischer, histopathologischer, ultrastruktureller und immunhistologischer Befunde gesammelt. Alle Pferde waren Quarterhorses oder von Quarterhorse-Abstammung.
Stammbaum-Untersuchungen gaben starke Hinweise auf autosomal rezessive Vererbung. Die hufigsten
Lsionen waren Serome/Hmatome, offene Wunden, Hautverlust und lockere, leicht abziehbare Haut, die nicht
in ihre ursprngliche Position zurckkehrte. Die definitive Diagnose konnte nicht durch Histopathologie gestellt
werden, obwohl das Vorhandensein von dicht gruppierten dnnen und verkrzten Collagenfasern, die in der
tiefen Dermis in Clustern angeordnet waren, auf die Erkrankung hinweisend war. Trichrome-, Orcein-Giemsaund immunhistochemische Frbungen auf Collagen I und III zeigten im Vergleich zu den Kontrollpferden keine
konsistenten Abweichungen. Elektronenmikroskopie zeigte keine Abnormalitten in der Periodizitt der
Kollagenbndel; weder Orientierung noch Variation im Querschnittsdurchmesser der Kollagenfibrillen konnten
Kontrollpferde von befallenen Pferden unterscheiden. Die Diagnose von HERDA basiert auf dem klinischen
Bild, aber kann durch suggestive (obwohl nicht pathognomonische) histopathologische Lsionen untersttzt
werden.

2004 European Society of Veterinary Dermatology, Veterinary Dermatology, 15, 207217